Compatibility and stability of 8% amino acids solution in ...

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University of the Pacific Theses and Dissertations Graduate School

1973

Compatibility and stability of 8% amino acids solution in Compatibility and stability of 8% amino acids solution in

combination with electrolytes, vitamins and antibiotics combination with electrolytes, vitamins and antibiotics

David Harold Schuetz University of the Pacific

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COfJIPATIBILITY AND STABILITY OF 896 AMll'lO ACIDS SOLU'l'ION

IN COJ:v1BINATION \'liTH EIJECTROLJ:.'TES, VITANINS AND .ANTIBia~ICS

A Thesis

Presented to

the Faculty of the .. Graduate School . . . . .

University of the Pacific

\ . .

In Partial Fulfillment

of the Requirements for the Degree

Masters of Science

by

David Harold Schuetz

November 1973

This thesis, writt,en and submitted by

David Harold Schuetz

is approved for recommendation to the Committee

----------------------on-Graduate-Studies, -Univer·sity of the Pacific.-

Department Chairman or Dean:

Chairman

Dated. _______ ~No~v~e~m~be~r~7~·-:1~97~3~----------------

ACKNOWLEDGMENTS

The author wishes to express his sincere appreciation

through the following acknowledgments&

To Dr. James King, who provided the opportunity to

undertake this research. His guidance and suggestions as

committee chairman will always be remembered. - - - -- --

To Dr. James McDavid a·nd Cutter Laboratories, InE.,

who provided the research grant and materials utilized in this

investigation, I am especially indebted.

To Dr. Herschel Frye and Dr. Francis Sayre for their

interest, ready availability, helpful advice during the

experimental phase and reading of this work.

To Dr. Patrick Catania, sincere thanks are extended

for his "behind the scenes" assistance.

To my parents and in-laws. They have inculcated the

value of education and bore the brunt of its setbacks over

the many years with unfaultering encouragement.

And finally to Vicki, my wife, and Christopher, my

young son, this work is dedicated. Through their sacrifice,

understanding, and inspiration, the frustrations and temporary

setbacks evolved into a successful milestone.

University of the Pacific

Stockton, California

November 1973

D. H. S.

TABLE OF CONTENTS

· LIST OF TABLES • •

LIST OF FIGURES

• • • • • • • • • • • • • • • • • •

. . . . . . ~~- . . ~ . . . . • • • • • •

LIST OJ? GRAPHS • • . . . ~ . . . • • • • . . . .. • • • ~---------------

Chapter

1.· INTRODUCTION . . . . . . ~ . . . . . . . . .

2.

NUTRITIONAL REQUIREf·1ENTS AJJLEVI.ATEJ1 BY TOTAL PARENTERAL NUTRITION ••••• • •

Protein requirement ~ • • • • • • • .. . . EJ.ectro1yt;e requirement • • • • • • • • •

Vitamin requirement ~ .. • * ... .. . . .. IJ ..

TYPES OF INCOHPA~~IBILITIES • • • • • • • •

PR.OBLEJ.VI • • • • .. • • • • • " e • • • • :It 11

. . .. . .. • • "' . . . . PHYSICAL Cm1PATIDII~ITY STUDY • « • • • • •

CHEHICAL CGr·1PNriBILITY STUDY • • • • • • •

U.V. Spectroscopy •••• ~ . ... . "' .. . . Thin Layer Chromatography • • & • ~ . ... . l\1icrobiologica.l Assay ., • ~ .. . . . . . ..

:r:X:PERINFJTTAL RESUI1J~S ~ • 0 * 0 • • • • .. .. ..

l)HYSICAL C0l\1PATIBILITY ANALYSIS .... • • •

Electrolyte admixtures

Electrolyte and vitamin admixtures .. . ..

v

Page

vii

X

xii

1

i.~

5

5

9

14

J.L~

16

16

18

18

20

22

27

27

27

33

Chapter

l.J ••

Electrolyte, vitamin, and insulin admixtures • . .. .. • • • • • ... • . .

Ele·ctrolyt e ~ vitamin, and antibiotic admixtures • . . • • • • • . • •

CHEMICALCONPATIBILITY ANALYSIS • • •

U. V. Spectroscopy • • • • • • • • •

• • • • Thin Layer Chromatography •

Microbiological Assay • . . .. • • •

• • •

• • •

....

. .. .

. . . • • •

DISCUSSION • • • • • • • • • • • • • • • • •

5.· CONCLUSION • • • • • • • • • • • • • • • • •

BIBLIOGRAPHY • • • • • • • • • • • • • • • •

..

vi

Page

38

lj.J.

52

52

82

86

97

105

107

LIST OF TABLES

Table

1. The Ingredients of 896 Amino Acids Solution • •

2. The Vitamin Content of ~WI 10 ml Ampul, MVI 5 ml Multidose Vial, Solu B Forte 10 ml, and

Page

7

Berocca C 2 ml Ampul • • • • • • • • • • • • 11

. "

4.. Physical Compatibility of Potassium Phosphate and Calcium Gluconate Admixed. to Equal Parts· of 896 Amino Acids Solution and De:x:trose in Water 5096 • • • • • • • • .. • • • • • • • • •

5.. pH of Potassium Phosphate and Calciurn Gluconate Admixed to Equal Parts 896 Amino Acids · Solution and Dextrose in \1/ater 5096 • • .. .. •

6. Physical Compatibility and pH of Potassium Phosphate and I'-'fagnesj_um .Sulfate Admixed. to Equal I>arts 85'6 J'.1.mino Acids Solution and De .. .-l-I'O'~e -in T.Tater 50°o/

7.

1:... U ).:> ...t.. Y~ I u • • • e • • e dl • • e

Physical Compatibility and pH of Calc.:ium Gluconate and Nagnesium Sulfate Admixed Equal Parts 896 Amino Acids Solution and Dextrose in Hater 5096 • • • • • • • .. •

to

.. . "

8. Physical Compatibility and pH o.f Solu B Forte and. r,lVI I'Jith Various Concentrations of Electrolytes Admixed to Equal Parts 8)6 Amino

23 - ---

29

30

31

32

Acids Solution and Dextrose in Water 5076 • • 3L~

9. Physical Compatibility and pH of l!,o.l-v:Lte and Rubramin PC with Various Concentrations of Electrolytes .Admixed to Equal P::trts 856 .Am:ino Acids Solution and Dextrose in \Vater 5096 • • 35

10. Physical Compatibility and pH of Aquamephyton ·with Various Coneentrations of Electrolytes Ac1mixed to Equal Parts. 896 Amino Acids Solution and DexL:ro;::;e 5:n Water 5096 • .., , .. • 36

vii

viii

Table Page

II. Physical Compatibility and pH of Solu B Forte or MVI with Selected Vitamins and Various Concentrations of Electr6lytes Admixed to Equal Parts 896 Amino Acid·s Solution and Dextrose in 1tlater 5076 • • • • • • • • • • • • 37

12. Physical Compatibility of Iletin U-40 Added to Various Concentrations of Electrolytes and Vitamins Admixed to Equal .Parts 896 Amino Acids Solution and Dextrose in \'later 50?6 • • 39

13. Physical Compatibility of Iletin U-40 Adc1ed to Various Concentrations of Electrolytes and

------ --v-itamins-:Admixe·d- to Equal Parts 896 Amino Acids Solution and Dextrose in \·later 5096 • • 40

14.

15.

16.

17.

18.

19.

20.

Physical Compatibility and pH of Selected Antibiotics Admixed to Equal Parts 896 Amino Acids Solution and Dextrose in \'later 5096 • •

Physical Compatibility and pH of Selected Antibiotics Admixed to V:::trious Concentrations of Electrolytes in a Hixtu.re of Equal Parts 8% Amino Acids Solution and Dextrose in it! at er 50% • • • • • • • • • • • • • • • • • ~ ~ • •

Physical Compatibility and pH of Selected Antibiotics AdJnixed to Various Concentrations of Electrolytes and Vitamins in a Nixture of Equal Parts 896 Amino Acic1s Solution and D xt · l·J t 50c:! e""" rose J..n ,a er ;o .............. .

Physical Compatibility and pH of Selected Antibiotics Admixed to Various Concentrations of Electrolytes and Vitamins in a r1ixture of Equal Parts 8% Amino Acids·Solution and Dextrose in 11Vater 5096 • • .. • .. • • • • .. • .,

Inhibition of Grovvth of Btanh~.£.~~ §]!}:'e·"L~s. by Pol;;rcillin N at a Concentration of 20 ggTml in a Nixture of Equal Parts 856 Amino Acids Solution and Dextrose in vJater 50~'6 • • • • ..

Inhibition of Grm.,rth of Staphl_ococcus ~e-~s by Polycillin N at a Concentra.tion of 20 p,gTml in.Therapeutic Concentrations of Electrolytes Admixed to Equal Parts 896 Amino Acids Solution and Dextrose in \'later 5096 • • . .. • •

Inhibition of Gro\,rGh of §i§.l?11lococcus §.Ureus by Kef1in at a Concentration of 50 J·Vslml in a Hixt;ure of Equal Part; s 896 Amino Acids Solution and Dextrose in \'later 5096 • • • • o

42

43

49 .

88

89

90

ix

Table Page

21. Inhibition of Grm-rth of Staphlococcus aureus by Keflin at a COncentration of 50 M,g/ml in Therapeutic Concentrations of Electrolytes Admixed to Equal Parts 896 Amino Acids Solution and Dextrose in Water 50% • • • • • · 91

22. Inhibition of Grov-rth of Staphlococcus aureus by Kantrex at a Concentrat~on of 400 Ag/ml in a Mixture of Equal Parts 8% Amino Acids Solution and Dextrose in Water 50% • • • • • 92

23. Inhibition of Gro11rth of Staphlococcus aureus by Kantrex at a Concentration of LJ-00 ,4g/ml in

------------------TlTerapeut±c-Concentrations of -Electrolytes Admixed to Equal Parts 876 Amino Acids

24.

25.

26.

.Solution and Dextrose in Water 5096 • • • • •

Inhibition of Grmvth of Staphlococcus aureus by Garamycin at a Concentrat~on of 80 Mg/ml in a Iv1ixture of Equal Parts 896 Amino Acids Solution and Dextrose in Water 5076 .. • • • •

Inhibition of Grm,rth of Stanhlococcus aureus by Gararuycin at a ConcentratiOn of 80 Mg/ml in Therapeutic Concentrations of Electrolytes Ad.>nixed to Equal l1arts 8~6 Amino Acids Solution and Dextrose in Water 50% • • • • •

Calculated Results o:f "Student t" Evaluation ..

93

94

95

96

LIST OF FIGURES

Figure

1. Standard U.V. Spectrograms of Equal Parts 8% Amino Acids Solution and Dextrose in Hater

Page

50% • • • • • • • • • • • • • • • • • • • • 54

2. Standard U.V. Spectrogram of Solu B Forte (o.l xl/ml) • • • • • • • • • • • • • • • • 55

3. Standard u.v. Spectrogram of 1'-TVI (0.2 ffl/ml) • 56

4. ·Standard U.V. Spectrogram of Folvite (5 J.{g/ml) 57

5.

6.

8.

9.

Standard U.V. Spectrogram o.f Rubramin PC (15 Rg/ml) . • • • • • • • • • .... • • .. • •

Standard U.V. Spectrogram of Aquamcphyton (10 }\g/ml) • . • • • • • • • •

U.V.·Spectrograms of Equal Parts Acids Solution and Dextrose in Solu B Forte 10 ml/L, One Hour ,admixture • • • • • • • • • •

• • . • • • •

891; Amino VJ at er 50~b and Post-• • • • • • •

U. V. Spectrograms of Equal Parts 85'6 Amino Acids Solution and Dextrose in Hater 505'6 Solu B Forte 10 ml/L, :Four Hours Post--

and

admi:x .. ture . • . l.t • a • • • • • • • • • • •

U.V. Spectrograms of Equal P:1rts Acids Solution and De:xtrose in Solu B Forte 10 ml/L, 8 and 2L~ admixture • • • • • • • .. • •

85'6 .i-\.mino 'vlater 505'6 and Hours Post-o • e • • • •

10. U.V. Spectrograms of Equal Parts 8% Amino Acids Solution and Dex:tr·ose in Water 5096 and

58

59

62

63

6Lf-

HVI 5 ml/L, One Hour Post-admixture • • • • 66

11. U. V. Spectrograms of Equal P.:1rt s 896 Amino Acids Solution and DeA.rtrose. in \vat er 5096 and HVI 5 ml/L, Four Hours Post-admi::-...rture • • • 67

12. U. V. Spectrograms of Equal Parts 896 Amino Acids Solution and Dextrose in \vater 5076 and MVI 5 ml/L, Eight Hours Post-admixture • • • 68

X

Figure

13. U. V. Spectrograms of Equal Parts 896 .Amino Acids Solution and Dextrose in \·later 5096 and

xi

Page

MVI 5 ml/L, 2LJ- Hours Post-admixture • • • • 69

14.

15.

16.

17.

18.

19.

20.

. 21.

22.

23.

2LJ- ..

25 ..

Standard u.v. Spectrogram of Polycillin N (0.8 mg/ml) • • • • • • • • • • • • • • • •

Standard U.V. Spectrogram of Keflin (20 J.tg/ml)

Standard U.V. Spectrogram o'f Garamycin (200 J.m/ml) . • . • • • • • • • • • • . . •

Standard U.V. Spectrogram of Kant rex (4 mg/ml)

8% Amino U.V. Spectrograms of Equal Parts Acids Solution and Dextrose in Combination v1i th Polycillin N, 12 Hours Post-admixture • • •

Vi at er 5096 in 1 Gm/L, 1 and • • • • • • •

u.v. Spectrograms of Equal Parts 896 Amino Acids Solution and Dextrose in Hater S0°6 , I and Keflin 2 Gms/L, One Hour Post-admixture • •

u.v. Spectrograms of Equal Parts 8% Amino Acids Solution and Dextrose in \'later 5096 and 'Keflin 2 Gms/L. 12 Hours Post-admixture " •

U.V. Spectrograms of Equal Parts 8% Amino Acids Solution and Dextrose in Water 509•6 in Combination with G:'lramycin 80 mg/L, 1 and 12 Hours Post-admixture • • • . • • • • • • • • •

U # V. Spectrograms of Equal Parts 896 Amino Acids Solution anc1 Dextrose in Water 5096 in Combination "~dith KJ.ntrex 500 mg/L, 1 and 12 Hot1.rs Post-admixture • • • • • ~ • • ,. • • •

Standard Chromatogram of Equal Parts 896 lunino Acids Solution and Dextrose .in \vater 5096 (l:J_) • • • • • • • • • • • • e • • • • • •

Chromatogram of Equal Parts 896 Amino Acids Solution and Dextrose in Hater 5096 Containing Potassium l~osphate 20 mEq/IJ, Calcium Gluconate 10 mEq/IJ, Solu B Forte 10 ml/L, :8'olvite 5 mg/L, Rubramin PC 1 mg/L, and Aquamephyton 10 mg/L One and 12 Hours Post-admixture • • • • ~ • • • • • • • • • •

Chromatogram of Equal Parts 8?6 Amino Acids Solution and Dextrose in 1dater 5096 Containing Potassium Phosph3.te 20 mEq/L, Calcium Glucon3.te 10 mEq/IJ, MVI 10 ml/L, Folvite 5 mg/L, R.ubramin PC 1 mg/L, and Aquamephyi;on 10 mg/L One and 12 Hours Post-aclmixture • .. • .. • • ., .. •. • .. .. ., • • • •

72

73

74

75

77

78

?9

80

81

83

84

85

LIST OF GRAPI:IS

Graph Page

1. Standard Beer's La\v Curve for Equal Parts 896 Amino Acids Solution and Dextrose in Water 5096 • • •. • • • • • • • • .. • • • • • • • • • 54

2. Standard Beer's J.Ja\v Curve for Solu B J?orte 10 ml/L • • . • • • • • • • • • • • • • • • • 55

----------- ------------- --- ----

3. Standard Beer's La~H Curve for r~wr 10 ml/I, • • • 56

4. Standarcl Beer's LavJ Curve for Folvite • .. .. • • 57

5~ Standard. Beer's Lav1 Curve for Rubramin PC .. • • 58

6. Stnndard Beer's Law Curve for Aquamephyton • • 59

7. Standard Beer's J;avJ Curve for Polycillin N • • 72

8. Standai·d Beer's Law Curve for Ke:flin . • • .. • 73

9 .. Standard Beer's Law Curve for Garamycin • • • .. 7l~

10. Standard. Beer's La\·I Curve for Kant rex ., • .. . t) 75

xii

Chapter 1

INTRODUCTION

For centuries, researchers have attempted to devise

the ideal .parenteral nutritional product readily amenable to

physiological requirements. Hyperalimentation, intravenous

alimentation, parenteral alimentation, parenteral feeding,

and total parenteral nutrition are synonyms which refer to

a method of complete intravenous nutrition reserved for

patients demonstrating negative nitrogen balance.

--------- ---

Substances such as wine, h.oney, and oils found common

employment in intravenous nutrition from the seventeenth to

nineteenth century, prior to the development and refinement

of lipid emulsions (1). The lipid preparations possess

greater nutritional energy, yielding 9 calories/Gm as com

pared to approximately 4 calories/Gm from dextrose (2). .HovJ

ever, limiteo. success was attained in this country in 1935

with parenterally administered lipid emulsions (3) .. The prob-·

lem of volume overload, a common occurrence when utilizing

dextrose vehicles, was reduced. The employment of fatty acid

preparations rapidly revealed numerous unpleasant·side effects

related to excessive fat metabolism (3). Patients complained

of nauGea, vomiting, chills, and symptoms of what became

know"Il as the "colloid reaction". The latter response was

manifested by back pain, flushing, cyanosis, apprehension,

1

2

dyspnea, and was occasionally accompanied by hemorirhaging .. ·.

tendencies, fever, epigastric pain, jaundice, thrombocytopenia,

and lipid deposition inthe Kiipfer·cells (3-5). Lipid emul

sions were obviously. not .the panacea of parenteral feeding.

Parenteral administration of amino acids by Elman (6),

in 1938, provided encouraging results. However, the initial

breakthrough in total parenteral feeding was accomplished by

Dudrick, et al. (7,8) in 1966. This proced.ure was soon modi-

fied for clinical use after demonstration of normal weight

gain and positive nitrogen balance in experimental animals

(9-11). Dudrick's formulation consisted of a hyperosmolar

infusate containing protein hydrolysates, dextrose, electro

lytes, vitamins and minerals. administered via an indv!elling

catheter positioned. in the subclavian vein (1,4-,9,12).

Protein Hydrolysate Injection, U.S.P., remained the

principle source of nutritional nitrogen until just a fe'iv

years ago. This product ,.,as prepared initially from hydro

lyzed casein and subsequently from beef blood fibrin. HovT

ever, the hydrolysates required the addition of cystine and

tryptophan after processing for pyrogens (3). Unfortunately,

they contained non-metabolizable polypeptides, which con-

tributed to significant water and sodium retention, positive

potassium balance, zero nitrogen balance, and ammoniuria (12).

Those cases of hypersensitivity to protein hydrolysates c.ited

were postulated to be related to the.polypeptide content (15,

16)~

The inherent disadvantages of protein hydrolysates

;- -'-'.=--=-=--'-.-'--=----

prompted·researchers to undertake an investigation employing

crystalline or synthetic amino acids as a nutritional source.

It was determined that these preparations would offer purity,

lack of non-essential amino acids,· flexibility of individual

component concentration, reduction of BUN levels due to urea

utilization in protein synthesis (1B,37), and decreased.

instances of anaphylactoid reactions (3). Flexibility of

ingredient concentrationwill permit future amino acid·solu-

tion formulations to correspond better to true physiological

plasma concentrations, \vhich is a concern expressed by Stegink,

et al. (18). Unfortunately, metabolism of DL amino acid. com

binations was found incomplete (13). These preparations

appeared to promot~ increased urinary urea levels accompanied

with negative nitrogen, potassium and sodium balance (12). On

the other hand., synthetic L amino acid formulations were fo1md

superior, especially vvhen administered to patients v1ith acute

renal failure (15-17).

Parenteral alimentation has been indicated for patients

exhibiting a catabolic state resulting from decreased caloric

intake or increased caloric requirement (19). As catalysis

progresses, cachectic patients' ability to recover and ward

off further infection is reduced (3,10).

Clinical indications for hyperalimentation include (20):

1. Patients "v'Jho cannot eat: esophageal carcinoma gastric carcinoma obstructive peptic ulcer paralytic ileus

2.. Patients \'Jho should avoid eating:

traumatic or inflammatory enterocutaneous fistulas affliction

regional enteritis granulomatous colitis pancreatitis laryngeal incompetence

l,L

3. Patients who cannot consume sufficient foodstuffs: multiple injury affliction major full thickness burns ulcerative colitis short bowel syndrome malabsorption syndrome

4. Patients who refuse to eat sufficient amounts: ----------------po-st- -operat-ive- --geriactric s-

anorexia nervosa, etc.

·5. Infants with congenital anomalies or chronic· diarrhea

Many complications evobring with the implementation·

of hyperalimentation are possibly due to technique rather than

the procedure. There have been reports of thrombosis of major

Yessels (i), sepsis (!+,11,22--26,35,45), extravasation (22),

hyperglycemia associated \'lith osmotic diuresis, dehydration,

and hyperosmolar state (14,15,22 ,2'7-29), volume overl0ad and

heart failure (1,22), metabolic acidosis (22,30-32), abnormal

plasma aminograms (22), bone abnormalities (1,22,33,34) hypo

phosphatemia (22,28,36,37), liver necrosis (1,22), vitamin and

trace mineral deficiencies (1,22,3L~,36,37), hypersensitivity

to protein hydrolysate (14,15), possible cholestasis (38),

hyperammonemia and hyperaminoacidemia (30,39,76), glucosuria

(14-), occurrence of essential fatty acid deficiency (40), and

iatrogenic hypercalcemia (1+1).

Complete parenteral feeding necessitates a balanced

infusate containing protein, carbohydrate, electrolytes,

vitamins, and minerals adapted to patient requisites (1,1+,9-12).

5

To mai:trliain positive nitrogen balance, approximately 150

non-protein calories are required for each·Gm of protein.

nitrogen (1,19,43). Proper correlation of caloric intake

with protein will induce the most efficient anabolic.state

(45), as ·decreased intake of non-protein calories may promote

protein catabolism to fulfill energy requirements resulting

in decreased nitrogen balance (42). Each 100 Gms of protein

will yield approximately 14 to 16 Gms of nitrogen (19,44).

]•or the normal adult at optimum metabolic ·efficiency, Munro

stated that daily body requirements could be estimated

probably not to exceed 0.45 Gms nitrogen/Kg body weight (46).

Severely burned patients may demand up to 10 times this

q~antity (3~43). The remaining sl~pplemental additives and

concentrations utilized depend upon patient need determined

through regular serum level analysis (4,10,47).. Consequently,

discussion concerning these additives, rationale for inclusion,

therapeutic levels utilized, and compatibility problems

encountered, will serve as the criteria in establishing the

guidelines for the investigation of compatibility and stability

of 8% Amino Acids Solutiona (see Table 1) in the presence of

supplemental admixtures.

The requirement for mono-valent electrolytes has, been

well documented (4,5,11,12,18,19,32,L~5,48-58). Sodium ion is

required in the development of membrane potentials and the

· activation of the cellular membrane at the onset of action

a- Trademark, Cutter Laboratories, Inc., Berkeley, Ca.

___

potentials (59). In addition, this cation influences the

tonicity of body fluids and controls the osmotic pressure

6

of the extracellular fluid (60) •. · \Dudrick, ~ §;1. (8,10)

have suggested 125 to 150 mEq of sodium supple.mentation

over a 24hour period to assist in promoting ·positive nitro-

gen balance.

Potassium has been reported to render significant

roles in the conduction of nerve impulses, muscular contrac-

tion, enzymatic reactions, and cell membrane potentials (61).

The dally patient regimen has required 75 to 120 mEq in main

taining physiologic function and promoting positive nitroge!.l

balance (8,10).

Complete intravenous alimentation will necessarily

.dmta5.J. the actmixture o.f various poly-valent electrolytes in

accordance 1.vith patient serum levels (1,3-5,10-12,18,19,45,

47-57,62). Calcium does play an important role in the physio

logical body functions being an ion essential for the skeletal

system, regulation of cell membrane permiability and resulting

action potentials, muscle contraction, release of ADH from the

neurohypophysis, release of catecholamines from the adrenal

medulla., blood coagulation, normal cardiac function, and

maintenance of the continuum (63). Therapeutic calcium requi

sites cited are dependent upon the age and affliction of the

pa·tient. .Neonates and infants demand higher levels due to

their elevated grovrth rate. Some authorities suggest 0 .. 5 to

4 mEq/Kg/2~- hours (18,48,49,55,56), others 2 to 25 mEq/L of

infusate (12,19,50,51,53). Adult concentrations may vary

7

Ingredients of 8% Amino Acids. ~Solution/100 ml

*Essential amino acids

Aminoacetic Acid, N.F ••••••• L-Arginine • • • • • • • • • • • • L-Aspartic Acid • • • • • • • • • L-Glutamic Acid • • .. • • • • • • • I.~-Hi-st-i-d-i-n-e--.-----.--.---.--.---.---- • • -. • ---. L-Isoleucine* • • • • • • • • • • L-Leucine* • • • • • • • • • • • • L-Lysine HCl* •••••••••• L-Methionine* • • • • • • • • • • L-Phenylalanine* • • • • • • • • • L-Proline • • • • • • • • • • • L-Threonine* .. • • • • • • • • • • L-Tryptophan* • • ••••••• L-Valine* • ~ ~ • • • • • • • • •

• • •

• • • • • • • • • •

• Sodium Ac et ate 'I'rihydrat e , N. ]'. Potassium Aceta·t::-;, N .F. . o • • • •

:Potassium Chloride? U.,S.P., ...... Magnesium Chlorid.e Hexahydrate • • Potassium fvietabisulfite ...... Water for Injection, qs •••••

..

.. • •

•

• • • • • • • • • • • • • • • • . - . -. . • • • • • • • • • c • •

• • • • . . . -· • • • • • • • • • • • 0

• • • • • • • • • • • • . . .. • • c: •

• • • • • • • •

Electrolytes Provided By

• • 3.387 grams • • 0.749 grams • • 0.400 grams • • 0 .l.f-26 grams

• • • • •

• • • • • • • • •

-- • 0. 237 -grams- -- __ • 0.493 grams • 0. 3L~7 grams • 0.667 grams •. 0. 427 grams o 0. L~OO g-.cams • 0 .. 10'7 grams • 0.,160 grams • 0.080 grams • 0.253 grams • 0.388 grams • 0.211 grams • 0 .. 056 grams • 0.061 .. 0 .. 010 • 100

grams grams ml

One Liter of 8% Amino Acids Solution

Sodium •• Potassium Magnesium Chloride .. Acetate •

•

• .. ..

• • • • 0

• • • • • • • • • .. . • .. . •

• • • • • • • • • • • • • • • • • • •

• • • • • • • • • .. • .. • • • • • • • •

• • • • • .. . • • . .. • • ~ . • • • • •

• • .. •

• • • • ..

40 mEq 30 mEq

6 mEq 50 mEq 50 mEq

••

8

from 2 to 20 mEq/L (11,12,18,32,47,51,53,54,56-58). Calc$um

gluconate injection appears to be the preparation of choice

as the supplemental adjunct for calcium deficiencies (11,12,

45,47,51,53,57). However, problems \vith precipitation have

been noted 'I:Jith its addition to protein hydrolysate alimenta

tion solution (64,65), and the presence of sulfates, phosphates,

and alcohol (66) •

. Phosphate has an essential function in bone metabolism,

transformation and production of high energy compounds required

in the biochemical path1.vays (37 ,51), plasma buffering and renal

excretion of hydrogen ion (67). Routine serum level deter-

minationo will dictate patient need for supplementation.

I~fants may require 2 to ll.J. mEq/Kg/24 hours (18,45,'+8,55,56)

or 3.9 to 15 mEq/JJ (19,1+7,52,5LJ·). Upper limits as high as

25 mEq/650 ml of ;i.nfusate have been employed (12,53).. Supple

mental phosphate for adult hyperalimentation solutions may

range from 4 to 27 mEq/L (11,12,18,32,47,50,51,53,56,58).

However, the addition of phosphate to parenteral alimentation

solutions may lead to precipitation in combination with cal

cium gluconate (66).

Magnesium is required for various intermediate meta

bolic path~Jays for the production of energy from glucose for

protein synthesis (18,37), activation of the oxidative phos

phorylation enzyme systems (51), and membrane ATPase (63).

Daily observance of serum levels serve as a guidE-) in main

taining the physiological demand. Pediatric concentrations

utilized have varied from 0.5 to 2.0 mEq/Kg/21~- hours (19,48,

9

49,55,56), 1.2 to 1.8 mEq/IJ (50,54,56), and 10 mEq/650 ml

of infusate (53). Adult levels have varied from 2 to 8 mEq/L

(11,12,32,LJ.5,51,53,57 ,62). Magnesium sulfate injection appears

to be the· soluble salt form used most often (5,11,12,32,45,51,

53,57,62). As such, precipitation in the presence of calcium

gluconate (66), protein hydrolysates (65), alcohol, and phos

phates (68) has been reported. The manufacturer of 8% Amino

·Acids Solution elected to add magnesium chloride as the

hexahydrate to avoid the possibility of precipitation,

apparently linked with the sulfate salt form.

Trace elements may be required upon detection of

deficiency states. Recent literature describes copper

deficits associated with long term alimentation (34,69,70).

,,Such complications must be considered when administering

protein sources lacking trace elementso The employment of

2 to 3 mg of iron dextran is useful in relieving anemias

(4,11,18,47) •. Supplementation of other trace minerals such

as zinc, cobalt, manganese, and iodine has been implemented

through intermittant administration of plasma or 1r1hole blood

when the necessity arises (1,4,8) ..

Physiological demand forvitamins appears undisputed.'

Patai remarked that ". • • B-complex vitamins all containing.

nitrogen appear to function, in a more or less understood

way, to act as cofactors for enzyme-catalyzed metabolic

reactions~ •• " (73). Greene (78) has implied that:

1. Vitamins are intimately involved in metabolism of carbohydrates, protein, and fat. As a result, they definitely possess a role in parenteral nutrition.

2. Due to rapidly developing deficiencies in patients considered for parenteral nutrition~ their inclusion is mandatory.

3. The requirement for parenteral vmter soluble vitamins appears magnified as compared to oral demands due to renal excretion.

4. Whenever fat soluble vitamins are employed; care should be exercised to avoid overdosing the patient.

10

Vitamin combinations containing fat and water soluble

componen.tsa have received the greatest utilization (1,4,5,11,

18,45,50-52,57,58,62). Employment of B-complex with ascorbic

acidb has also been mentioned (18,58,71). Infant requirements

for fat and water soluble vitamins have been fulfilled with

1-10. ml of NVI (1,18,19,L~5,52,72). Adult requisites were met

with 5-10 ml of MVI (L~,5,11,18,50,51,57,58,62), 10 ml of

Solu B Forte (58,71), and 2 ml of Berocca C (18). Refer to

Table 2 for vitamin content.,

Folic acid participates in the conversion of certain

amino a.cids to intermediates in production of deoxyribonucleic

acid (7~-)~ Consequently, this vitamin remains essential for

normal grov~h and maturation of erythrocytes (74). Therapeutic

supplementation may vary from 5 micrograms to 10 mg (1,4,11,32~

45,LV?,50,51,57). The addition of folic acid to nutritional

formulashas presented compatibility problems .. Burke stated

precipitation may occur in the presence o.f calcium salts (79).

a- I1VI, u.s.v. Pharmaceutical Corp., Tuckahoe, N.Y.

b - Solu B Forte, The Upjohn Co., Kalamazoo, Mich.; Berocca C, Roche I1aboratories~ Div~, Hoffmann-La Roche, Nutley, N •. J •

Table 2

Vitamin Content of MVI 10 ml Ampul and

MVI Concentrate 5 ml Multidose Vial

11

Vitamin A • • • • • • • • • • • • • • • Vitamin D • • •. ., • • • • • • ••• ~ • Vitamin E (DL alpha Tocopheryl

10,000 u.s.P. units 4,000 U.S.P. units

acetate) • • . • • • • • • • • • • • 5 International Thiamine_H_QL_~.--• __ • .. • • _.· __ • _. • .. • • • • _ • • __ • _. Riboflavin • • • • • • • • .. • • • • • • • • . • • • Niacinamide • • • • • • • • • • • • • • • • • • Pyridoxine HCL • • • • • • • • • • • • • • • • • • Dexpanthenol • • • • • • • • • • • • • • • • • • • Ascorbic Acid • • • • • • • • • • • • • • • • • •

Vitamin Content of Solu B Forte 10 ml

Thiamine HCL • .. • • • • • • • • • • • • • • • • • Riboflavin • • • • • • • • • • • • .. • • • • • • • Pyridoxine HCL • • • • • .. • • • • • • . • • • • •

units 50 mg -io mg ---- - -

100 mg 15 mg 25 mg

500 mg

250 mg 50 mg 50 mg

Niacinamide • • • • • • • • • • • • • • • • • • • 1,250 mg Sodium Pantothenate • • .. • • • • .. • • • • • • .. Ascorbic Acid • • • • • • • • ~ • • • • • • • • •

Vitamin Content of Berocca C 2 ml Ampul

Thiamine HCL • e • ., • • ., • • .. • • • • • • • • •

Riboflavin • • • • • • • • • • • • • • • • • • • • Niacinamide • • • • • • • ~ • • e • • • • c • ., •

D-panthenol • • • • • • • • • • • • - • • • • • • • Pyridoxine HOL ., .. • • • ~- • .. .. • • • • • • • • • Ascorbic Acid • • • • • • • • • • • • • • • • • • d-Biotin • .. • • • • • • • • • • • • • • • • • • •

500 mg 1,000 mg

10 mg 10 mg 80 mg 20 mg 20 mg

100 mg 0.2 mg

12

Exposure to excessive light (80,81}, riboflavin (81), and

oxidizing or reducing agerits such as ascorbic acid nave been

implicated in the decomposition o:r· folic acid (80,81).

Cyanocobalamin is referred-to as the "Maturation

Factor" (76). As such,· Guyton contends it ". • • is· an

essential nutrient for all cells of. the body, and grmvth

of tissues in general is greatly depressed \vhen this vitamin

is lacking." (76). Clinicians continue to employ 1 to 1,000

micrograms of cyanocobalamin injection to fulfill the physio

logical· requirements during total parenteral nutrition (1,5,

11,12,32,45,47,50,51,57,58,71). However, inclusion of

cyanocobalamin in hyperalimentation solutions has been ques

tioned. Deterioration in the presence of ascorbic acid (79,

82), vitamin K (79), and elevated levels of dextrose (79) has

been reported.

Periodic administration of phytonadione has prevented

acute episodes of hypoprothrombinemia associated v.rith neonates

and patients exhibiting deranged gastrointestinal function or

flora (1). Therapeutic regimens again may vary from 5 micro

grams to 10 mg (1,18,32,45,47,50,51,57). Phytonadione readily

decomposes in the presence of light (80-82). There may also

be inactivation in combination with cyanocobalamin and ascorbic

acio. (64, 79).

Hyperglycemic states may be experienced with the onset

of hyperalimentation infusion. Attempts to control this unde-

sirable complication, especially for .diabetic patients, ·have

been undertaken with concomitant administration of insulin

13

injection (5,11,15). \Vyrich, et al. (15), employed C1initesta --determinations ot glucosuria as criteria for insulin supple

mentation. Fifteen units are administered for a +4 urine,

10 units for a +3 urine, and none·«for lower levels of gluco

suria. Regulation of infusion rates remains the best preven

tative control measure against hyperglycemic crisis. Serum

blood sugar levels above 500 mg 96 should not be permitted(15) ..

Prolongedperiods o.fhyperglycemia predispose patients to

complications of non-keto acidotic hyperosmotic coma (14,15,

22,27-29).

Burke has reported that some investigators indieateQ.

a 23-27% loss of insulin to glass and tubing adsorption (79).

Ho\vever, Abel, et al. (83) disclosed insulin adsorption, as

determined by radioimmun.oassay techniques, appears about 2%.

The results of a radio-assay analysis conducted by Weisenfeld,

et 2-.:l• (84) appears to coincide \'lith data mentioned by Burke

(79).

Clinicians continue to make inquiries concerning the

admixture of selected antibiotics to hyperalimentation solu

tions. Hyman, et al~ (45), incorporated 100,000 units of

potassium penicillin G into alimentation solutions for pre

vention of septicemias associated with bacterial contamination.

Inherent complications arise with such admixtures.. The com

patibility o.f antibiotic salt forms and the role of pH changes

have a great influence upon the feasibility of these

a - Clinitest, Ames Co., Inc .. , Elkhart·, Ind.

14

combinations (77,85).

Utilization of the purest source.of protein nitrogen

appears advantageous. This recent literature survey pertained

primarily to ingredients commonly admixed to protein hydro

lysates or modified fibrin hydrolysates. The compatibility.

and· stability of synthetic L-amina acids solutions in com

bination with customary additives have not been investigated

except for an electrolyte compatibility study by Kaminski,

et al. (86) on FreAmine 8.5%a.

·Incompatibilities are frequently classified as physi

cal, chemical, and therapeutic. Physical incompatibilities

are reported as visual manifestations of chemical interaction,

· i.~ e., precipitation, pH alteratioD:, color change~ or evolution

of gas (85). Chemical incompatibilities occur without visual

demonstration and can produce undesirable complications (85).

Therapeutic interactions are manifestations of directly

opposed pharmacologic action of two or more simultaneously

administered medications. This type of incompatibility will

not be considered in this discussion.

The 896 Amino Acids Solution is presently classified

as an investigational product. Consequently, information

regarding its compatibility and stability \vith commonly

utilized additives is relatively non-existent. The present

study was designed to observe a solution of amino acids in

combination with v·arious concentrations of previously described

a - Trademark, McGaw Laboratories, Glendale, Ca.

15

.adjuncts for evidence of physical compatibility. ·The

compatibility data resulting from that segment of the study

were thenutilized as a guideline in the determination of

chemical compatibility employing instrumental analysis.

Initially, u.v. spectroscopy was employed for the_

determination of chemical compatibility and stability of 8%

Amino Acids Solution in those admixtures determined to be

physically compatible. Those combinations ascertained as

compatible through U. V. spectroscopy \'Jere subjected to

further examination by thin layer chromatography. Finally,

those antibiotic/amino acids mixtures found physically com

patible \llere subjected to microbiological assay for the

-detection of possible inactivation or degradation of the

antimicrobial agent.

Chapter 2

EXPERIMENTAL METHOD

In. order to simulate clinical conditions, the

compatibility to the amino acids/dextrose solution admix

tures \'las determined under conditions of normal lighting

and at room temperature. Prior to the addition of the

various additives, the 8% ~nino Acids Solution was mixed

with equal portions of 50% dextrose in watera to form the

basic hyperalimentation solution. Aliquots of that mixture

s.~rved as the base to which other _ingredients were added,

to yield final concentrations as indicated.

£11-;y:sical Cogp_at];.bilit;y Study

To determine the physical compatibility characteris

tics of the basic amino acids/dextrose hyperalimentation

solution} various concentrations of the follo·wing additives

were added to the stock mixture.

Electrolytes b Potassium Phosphate Concentrate Calcium Gluconate lO%C Magnesium Sulfate 10%d

a - 509G Dextrose Injection, U.S.P., Cutter Labs. 5 Inc., Berkeley, Ca.

b -~ McGaw Laboratories, Glendale, Ca~

c- Calcium Gluconate Injection, U.S.,P., 10%, Pasadena Hesearch Laboratories, Pasadena, Ca.

d- Magnesium Sulfate Injection, U.S .. P., 10%, Eli Lilly and Co., Indianapolis, Ind.

16

Vitamins (alone and ·with physically compatible concentrations of electrolytes only)

MVI Conc€mtrat e Solu B Forte Folvitea Rubramin PCb Aquamephyton°

Insulin (with compatible combinations of vitamins and electrolytes)

Iletin U-4od

Antibiotics ------ ----------(aJ:orre-,-w±th-electrolytes, and with electro...;

lytes and vitamin combinations) Polycillin Ne Kantrexf Kefling Garamycinh

17

Each series of admixtures, examined for physical incom-

P.<?ttibility, was performed utilizing duplicate 25 ml samples of'

the stoek sol uti on~ As applicable, depe:nd.ing upon the number

a- Folic Acid Injection, Lederle Laboratories, Div., American Cyanamid Co., Pearl River, N. Ye

b- Cyanocobalamin Injection, U.S.P., E8 R. Squibb and Sons, Div .. , Olin f-'lathieson Chemical Corp., Nm·l York, N. Y.

c- Phytonadione, U.S.P., Merck Sharp and Dohme, Div., Merck & Co., \'lest Point, Pa.

d- Insulin Injection, UoS.P., Eli Lilly & Co., Indianapolis, Ind.

e - Sodium A..mpicillin for Injection, Bristol Laboratories, Div.~ Bristol Myers Co., Syracuse, N.Y.

f- Kanamycin Sulfate Injection, U.S.Po, Bristol Laboratories~ Div., Bristol Myers Co., Syracuse, N. Y.

g- Sterile Sodium Cephalothin, U.S.Pe, Eli Lilly & Co., Indianapolis, Ind.

h- Gentamicin Sulfate, U.SoP., Schering Corp., Bloomfield, N .. J.

of additives, the following order of.mixing was used

throughout this study, including physical, chemical, TLC,

and antimicrobial tests.

Potassium Phosphate Calcium Gluconate Magnesium Sulfate MVI or Solu B Forte Folvite Rubramin PC Aquamephyton Iletin U-4-0

biotic was added after the MVI or Solu B Forte.

18

The resulting admixtures, except where indicated,

were examined at 1, 4-, 8, and 24 hour intervals, against

black and \vhite backgrouno.s for evidence of physical change.

A. Corning pH I•leter I-1odel ?a was usea. to take readings at 1

and 24· hours after admixture, except i'There other\vise indicated.

Those samples failing to demonstrate physical change '"ere then

exam:i.ned for chemical compatibility.

9P.eE.J.i9..Q.l_Q..Q!gpat~biltty_Stud_;z

u. V • .Sr?ec..t.:roscopy.. For purposes of examination utiliz

ing U.V. spectroscopy, a Bausch and Lomb Spectronic 600 Double

Beam Spectrophotorneterb linked with a Linear/Log Varicord 4-3

Recorderc v1as employed. All solutions for investigation t-iere

---------·--·--------a - Corning Scientific Instruments, Medfield, Mass.

b - Bausch and Lomb Analytical Instruments, Rochester, N. Y.

c - Photovolt; Co:rp., New York, N. Y.

19

placed in Coleman 30-300 Silica Curvettesa .for the' scanning

pr·O<!edures. .A dueterium lamp furnished the energizing beam

for the solution$. in question in wavelengths-ranging from

220 to 320 nanometers. It was postulated that any alteration

of the speetrogram.s for ingredients in the ao.mixture as com.;

pared to their standard spectrogram in distilled \'Jater, would

indicate a chemical interaction.

The determination of suitable concentrations was

required for the production of consistent u.v. spectrograms

for each individual component. Individual additives were

diluted from their full strength status with double distilled

water. These samples \'!ere then scanned in comparison to a

r.~ference of double distilled wate.r. Once the U., V .. spectra

consistently measured an absorbance ranging .from 0 .. 2 to 1 .. 0,

Beer's law plots "~:le.re established for each component.

Following the preliminary inquiry, triplicate 25 ml

sample solutions containing .multiple additives were analyzed.

Using thE! Beer's la1:1 plots as a guideline, proper dilutions

were made, and U.V. spectrograms obtained for the desired

ingredient.. IJ.'he reference solution contained exactly the same·

components, at an identical concentration, excluo:ing that addi

tive being sGanned in the sample solution, Each sample was

subjected to examination at intervals of 1 7 '+, 8, and 2L~ hours

after ao...mixture. In o~cder to facilitate the investigation,

a - Coleman Instrurnents, Div .. , J?erkin-Elmer Corp., fvlayvwocl, T, 1 ••.• !. .L..

20

only those combinations of electrolytes representing "usua,l 11

therapeutic and highest physically compatible mixtures were

examinede Although some additive concentrations were found

too dilute, when possible, remaining ingredients \'lere examined

non·etheless for possible interaction.

Thin Layer Chromatography. As an adjunct to u.v. spectroscopy, TLC was employed to elucidate, further, possible

changes which may have taken place in the solutions.

Initially, a standard chromatogram for the amino acids/

dextrose solution \vas prepared. This \vas accomplished utiliz

ing a method of Frye (87) developed for the determination of

amino acid content in soybean products. Glass-supported, pre

c·oatecl cellulose TLC plates, 20X20 em, without fluorescent . . a ~ndl.cator ) were employed.. rrbey had a layer thickness of

0.10 mm& DrR~mond Microcapsb of 2 microliter capacity were

utilized throughout this investigation. After preliminary

trials, 0.5 microliter proved sufficient-to provide consistent

chromatograms. The spotted solution '11-Jas never allmrred to

diffuse more than 2 to 3 mm in diameter at the origin. This

was accomplished by applying small portions with intermittant

drying. A Desaga template0 proved beneficial in placing the

origin 1 em from the right edge and bottom of each plate~

Prior to each determination, the pre-coated plates ~trere

a- E. 11. IJaboratories, Inc., Elmsford, N.Y.

b - Dr-u.rnmond Scientific Co., U. s. A •

. c - Desaga, Heidelberg, West Germany.

21

developed in Sol vent System I, \'lhich was allowed ~o traverse

the entire surface before drying the plate.for 5 minutes at

100°C. Duplicate sample solutions were prepared .for each

determination.

Ground glass Desaga-Briru~manna developing tanks were

·utilized. Filter Paperb, 20.5 em in diameter, lined both

sides of each tank to insure saturation. Dual solvent systems

(Table 3) allowed the production of two-dimensional chromate-

--grams-.- Just prior to each determination, the old solvents

were replaced with freshly prepared solvents. The filter

paper 'ltlas changed simultaneously.

Prior to placing the plate in the first solvent, the

cellulose surface was etched across the entire coating 16 em ..

above the origin. Similarly, a second line was scored in

perpendicular.' fa;shion 17 em to the left of the origin to

insure a pre-designated. termination of the migrating second

solvent front. All plates 'ltlere developed while standing on

edge. Approximately 8 hours were required for the first

solvent to traverse the 16 em. The plates were removed,

dried at 100°C. for at least 5 minutes, rotated 90 degrees and

placed in the tallies containing the second solvento Development

time ranged from L~ to 6 hours.. The plates were again dried.

The drying proc;edures were necessary to insure removal of

traces of hydrochloric acid and ammonia ..

Visualization \vas accomplished through the· use of

a - Desaga, Heidelberg, West Germany.

b- Braun-Knecht-Heimann Co .. , San Francisco, Ca.

ninhydrin in a cadmium acetate solvent (Table 3). The

ninhydrin was admixed just prior to use of the spray

22

reagent .following the second drying cycle. The visualiza

tion solution was sprayed on the plates until they became

translucent. Care v.ras exercised to prevent the reagent .from

running. The damp plates were drie~ approximately 15 minutes

at 80°0.. Quite o.ften, more time was required at room tempera

ture .for distinct resolution .. The resultant.chromatograms

were traced on onionskin paper • . ·Having obtained consistent chromatograms .for the

amino acids/dextrose solution, duplicate samples of the

hyperalimentation solution containing electrolytes ana. vita

mins were examined 1 and 12 hours post admixture. Significant

changes i:q. spatial arrangement of the components on the chromato

grams, as compared to the standard, 'i:JOuld indicate possible

chemical interaction betv.reen the amino acids and additives.

Micr_opiolog~cal. Assay.. In order to verify possible

loss o:f antibiotic activity when admixed to the amino acids/

dextrose solution, selected antibiotics \vere added alone and

in combination with "usual" therapeutic concentrations of

electrolytes e It is the contention of the U .. S.P. (88), that

antibiotic efficacy is directly related to its demonstratable

abi1ity to j_nhibit microbial grOi\rth under proper conditions.

Loss of intrinsic antibiotic activity will manifest itself

through subtle changes inits capability to inhibit microbial

growth. This is exemplified by reduced zones of inhibition ..

The procedures set forth by u.s.P. XVII were modified

Table 3

Solvent System, First Dimension

2-Propanol • • • • • • • 'Butanone •••••••• lN Hydrochloric Acid • •

• • • • • • • • • • • •

• • • • • • • • • • • • • • • • • • • •• • • • • • • •

Solvent System, Second Dimension

2-Methyl-2-Butanol • • • • • • • • • • • • • • • • Butanone • • • • • • • • • • • • • • • • • • • • • P:r-opanone • • . .. • • • • • • • • • • • • • • • • • Methanol • • • ~ • • • • • • • • • • • • • • • • • Distilled \Vater • • • • • • • • • • • • • •••• Aqueous .Ammonia • • • • • • • • • • • • • • • • •

Cadmium Acetate-Ninhydrin Spray Reagent

Cadmium Acetate • • • • • • • • • • • • • • o • •

Distilled Water o • • • • $ • • • • • • .. • ••

Glacial Acetic Acid · • • • • • • • • .. • • • • • • hopanone , q .. s. • • • • • • o • • • • • • . • • • •

Ninhydrin (ado.ed just prior to spraying) •••••

23

60 parts 15 parts 25 parts

50 parts 20 parts 10 parts

5 parts 15 parts

5 parts

0.5 gm 50.0 ml 10.0 ml

500.0 ml 0.2 96 w/v

24

slightly, incorporation suggestioi:l.s of Grove and Randall (89).

The recommended agar cups were replaced with sterile, absorbant,

paper discs one-half inch (12.7 mrn) in diameter.a

Staphlococcus aureus, A.T·.;c.c., 25293, was chosen to

illustrate the antibiotic activity of Polycillin N, Kantrex,

Keflin, and Garamycin, when admixe~ individually with the

hyperalimentation solution. The culture vJaS maintained on.

nutrient agar slants consisting of Bacto Antibiotic Medium - - --

#lb. Bacterial transfers were performed weekly. Fresh

innoculated slants were incubated 24 hburs at 37°0. to stimu

late growth and then re.frigcrated. Nutrient broth was prepared

using Bacto Antibiotic Medium #2b. Both the agar and broth were

aut.oclaved before use and kept sterile for the remaining assay

procea.ures.

A day prim:· to the assay, all required materials v-mre

wrapped and autoclaved to insure sterility. Simultaneously,

a loop.ful o.f the bacteria was transferred from the agar slant

and dispersed in 2:.5 ml of sterile nutrient broth and incubated

24 hours at 37°0. Agar plates were prepared pipetting 25 ml

of freshly autoclaved Bacto Antibiotic Medium ill into sterile

disposable Lab-Tekc 100 x 15 mm plastic petri dishes.

At; the time of assay, the nutrient agar plates \vere

a- Schleicher and Schuell, Inc., Keene, N.H ..

b - Di.fco Laboratories, Detroit, Mich.

c - J~ab-Tek Products, Div.·, Miles Laboratories, Inc., Westmont, Ill ..

25

seeded with 0.1 ml of innoculated broth. The innocuJ.um was

spreac1. evenly about the surface of the agar with a L-shaped

glass rod and allowed to dry. The various solutions and

dilutions were then prepared utilizing Sterile Water for

Injection, u.s.P.a. Five absorbent discs were then positioned

equidistant about the dried agar surfaces of the petri dishes.

The sample solutions were spotted on the dry discs utilizing

50 lambda pipettesb. Following this procedure, the plates ---- -- - - -

were incubated 12 hours at 37°C. Longer incubation tended

to obscure the zones.. The zones of inhibitibn were measured

with a Fisher-Lilly Antibiotic Zone Readerc~

A preliminary study indicated 50 lambdas of Polycillin

N:_0.02 mg/ml, Kantrex 0 .. 4- mg/ml, K;eflin 0.05 mg/ml, and 100

lambdas of Garamycin 0.08 mg/ml would yield optimum zones of

inhib:i.tion for the investigation ..

Each of the four antibiotics was added to a 25 ml

portion of the total parenteral nutrition solution to produce

a concentration of Polycillin N, lGm/L; Kantrex, 500 mg/L;

Kefl.:i.n, 2 Gm/L; anc.1 Garamycin 80 mg/L. With the exception

of Garamycin whic.h was used undiluted., the antibiotic/

hyperali.mentation mixture \vas diluted to the optimum concen-

trations indicated above at one and 12 hours after admixture.

One hundred J.ambdas of the Garamycin solution and 50 lampdas /

a- Cutter Laboratories, Inc., Berkeley, Ca.

b - diSPo Capillary Pipets, T~M .. S/P, Div. AHSC ..

c- Fisher Scientific Co. 1 Pittsburg, Pa.

26

of each of the other solu·tions \'lere applied to the sterile

absorbent discs immediately after dilution. Fifteen repli

cate determinations were made for each mixture.

The same antibiotics, at the same concentration, were

added to the hyperalimentation solution to which 20 Meq/L

potassium phosphate and 10 mEq/L calcium gluconate had been

added. The results of the physical compatibility study indi

cated antibiotic admixture in combination with these supple

mental electrolyte concentrations demonstrated the greatest

degree of stability. These solutions were handled as outlined

above.

Antibiotics in sterile water and the hyperalimentation

solution without antibiotics served. as controls.. Following

incubation, the zones of inhibition v1ere ·compared with the

controls for differences in microbial inhibition ..

---- ----- ..... - -~--- -----------------

Chapter 3 · ···

EXPERIMENTAL RESULTS

Ph;y:sical Compatibility Analysis

Duplicate samples of.the amino acids/dextrose solution

were examined after various admixtures. The nature and con-

centration of these combinations are indicated on Tables L!. to

17. These mixtures were examined at the end of 1, 4, s; and

24 hours against black and white backgrounds for evidence of

physical change. The pH readings were taken 1 and 24 hours

after admixture. The absence of precipitation, color change,

significant pH aJteration, or evolution of ge.s suggested

physical compatibilit~r. The appearance of one or more of the

above indices, indicated incor.1patibility.

Part 1.. During the compatibility investigation of

the hyperalimentation solution and electrol;yte eombinations,

potassium phosphate was added ,before calcium gluconate. The

resultant concentrations found to be physically compatible

are indicated on Table 4o Those combinations of electrolytes

demonfJtrating immediate incompatibility usually appeared as

\<~hi te, milky solutions with fine particulate matter gradually

settling to the bottom of the flasks. Those incompatible

combinations bordering the compatible concentrations required.

12 to 24 hours for precipitation to 6ccur. In these cases,

the precipitate appearE:d as translucent or white flakes, most

27

28

o.ften .floating upon the sur.face o-f the solution.

In similar .fashion, magnesium sul.fate was examined

in the presence o.f potassium phosphate and calcium gluconat;e

contained in the basic stock solution (Tables 6 and 7). 'l'he

concentrations studied failed to demonstrate incompatibility.

It was also found that 160 mEq/L o~ magnesium sulfate could

be added to the hyperalimentation solution containing 40 mEq/L

o.f either potassium phosphate or calcium gluconate \vithout -·

evidence of physical change. However, since 8% Amino Acids

Solution already contains 6 mEq/L of magnesium chloride hexa

hydrate, furthel~ investigation including magnesium sulfate

addition was not undertaken.

The tabulated. pH readings in this part of the experi

mentation represent a mean of duplicate determinations. A

standard deviation is indicated in those instances where a

slight change in pH was observed.

8

6

4

2

29

Table 4

Physical Coinpatibility.of Potassium Phosphate : and Calcium Gluconate .Admixed to Equal Parts

of 8% Amino Acids Solution and Dextrose in Water 50%

--f--- ---1 . l I 1----r--+- ·.i

I

I c

C.

c J '----·----

3 6 9

C = Compatible

-

I I @

c c X X X -·

X

c c X X

c X I -

xj .. c c c X

c c c

c 0 c c

c

12 10 15 20 25 30 35 40

Potassium Phosphate (mEq/L)

X

X

. -

X

~-

.I I -- .• , I I I

! .,;

' ~--

I I

I I j

60 80 100

X = Incompatible @ = Precipitates unless agitated during mixing

-- --- - -------

1-·

6.,3

-~~-~- --

Table 5

The.pH of .Potassium Phosphate and Calcium Gluconate Admixed to Equal Parts 8% Amino Acids Solution and Dextrose· in \•later 5096

_,..._

I ------ -

I I I ., ' *

-- I

16.~ 6.~ I * 6 .. 3 6.3 ...... ....,.

I : I I *

.I * l

h ~- '-6. 3 !r;.LJ.. I . .

I I I 6.~ b 4.5+-i

. ;

s c:;s-1 IG 4 's ~s 6.S

6~5 6.55 -·

s _11 6...1L. h~ h.h

6.~

6.~

*

*

_j

I I I

" I

I

I

I I

!

! 3 6 9 12 10 15 20 25 30 35 ~0


60 80 100

* - Precipitated immediately upon acldition of calcium gluconate. No pH reading was taken ..

Amino acids/dextrose solution pH 6.15

Table 6

Physical Compatibility of Potassium Phosphate and Magnesium Sulfate Admixed to Equal Parts

896 Amino Acids Solution and Dextrose in Water 50%

·Magnesium 4 c c G Sulfate (mEq/L) 3 c c c c

2 c c c c ··-

1 l c c c c 10 20 30 40


The pH of Potassium Phosphate and Magnesium Sulfate Admixed to Equal Part's 896 Amino Acids

Solution and Dextrose in Water 50%

--Magnesium 4 6.35 6.45 6.55 6.60 Sulfate (mEq/L) 3 6.35 6.45 6.55 6.60

2 6.32 6.42 6.50 6.60

1 6.30 6.40 6.50 6.60 10 20 30 40


C = Compatible

Control Amino Acids/Dextrose Solution pH 6 .. 15

----------

Table 7

Physical Compatibility of Calcium Gluconate and Magnesium Sulfate Admixed to Equal Parts

8% Amino Acids Solution and Dextrose in \'later 50%

---- - - --

·Magnexium l.J. c c c c Sulfate (mEq/L) 3 c c c c

2 c c c c

1 I c c c c 10 20 30 1+0

Calcium Gluconate (mEq/L)

The pH of.Calcium Gluconate and Magnesium Sulfate Admixed to Equal Parts 8% Amino Acids Solution and Dextrose in 'dater 50%

Magnesium Lk 6.15 6.15 6.17 6.20 Sulfate (mEq/L) 3 6.15 6.17 6.17 6.20

2 6.10 6 .. 17 6 .. 17 6.20

1 6 .. 15 6.15 6e20 6.20

10 20 30 l.J.O

Calcium Gluconate (mEq/L)

G = Compatible

Control Amino Acids/Dextrose Solution pH 6.15

33

Part 2. The physical compatibility characteristics

of the electrolyte/amino acids solution were considered to

represent both "usual" therapeutic and highest compatible con

centrations. The succeeding investigation utilized these

categories of admixtures in addition to vitamins commonly

employed in total parenteral nutrition. In the absence of

other additives, all vitamin solutions ·tested appeared to.be

physically compatible vvith the basic amino acids/dextrose

solution .. The multiple component solutions were again observed

in identical fashion as those in Part 1. The :pH readings were

recorded similarly. They remained essentially unchanged

(Tables 8 to 10).

An examination involving a combination of vitamins

\'JaS und~?rtaken once the compatibility of' single vitamin aamix

tures to the electrolyte/amino acids solution was empirically

demonstrated (Table 11). Identical procedural techniques were

employed as previously described. These admixtures again

.failed to manifest any physical incompatibility.

------------- -

Solu

Table 8

Physical Compatibility and Corresponding pH of Solu B Forte with Various Concentrations

of Electrolytes Admixed to Equal Parts 8% . Amino Acids Solution and Dextrose in Water 50%

B Forte 10 ml/L c c c

Solu B Forte .5 ml/L I : c c c

Solu B Forte 10 m1/L 5-73 6.05 5.95 6.25 .:t.-0346

Solu B Forte 5 ml/L 6.00 6.25 6.20 6.40

Ca 10 Ca 20 Ca 25 Ca 15 p 10 p 25 p 20 p 40

*Control 6.40 6.45 6.40 6.55

Ph;tsical 0ompatibility and Corresponding pH of I"lVI with Various Concentrations of Electrolytes Admixed to Equal Parts 896

Amino Acids Solution and Dextrose in Water 5096

c

I c

5.90

6.15

Ca 40 p 15 6.30

------MVI 10 ml/L

MVI 5 m1/L

MVI 10 m1/J.J

MVI 5 ml/L

*Control

c c c c c

c c c c c

6.31 6 .. 45 6.40 6.51 6.36 .:t,.0264 .:t,.0264 .:t,.024

6.35 6.437 6.43 6o53 6.40 +.0264 .±_.0282 +.0282

Ca 10 Ca 20 Ca 25 Ca 15 Ca 40 p 10 p 25 p 20 p 40 p 15 6.40 6.45· 6.40 6.55 6.30

C = Compatible Ca = Calcium Gluconate in mEq/L P = Potassium Phosphate in mEq/L

*pH of the Amino Acids/Dextrose Solution with Electrolytes

-

Table 9

Physical Compatibility and Corresponding· pH of Folvite with Various Concentrations of. Electrolytes Admixed to Equal Parts 8%

Amino Acids Solution and Dextrose in Water 50%

]'olvite 5 mg/L c c c c

Folvite 2.5 mg/L c c c c

35

I c

c I --- - -- - ----- -- - - I Folvite 5 mg/L 6.33 6.1+0 6.40 6.45 6.35

±·0346

Folvite 2.5 mg/L 6.35 6.45 6.40 6.50 ,:t.070

.. Ca 10 Ca 20 Ca 25 Ca 15 p 10 p 25 p 20 p 40

*Control 6.40 6.4·5 6.40 6.55

Physical Compatibility and Corresponding pH of Rubramin :PC vd th Various Concentrations

of Electrolytes Admixed to Equal Parts 8% Amino Acids Solution and Dextx-ose in Water 50?6

6.33 +0.346

Ca 40 :p 15 6.30

==================================·--·--·---·----Rubramin PC 1 mg/L

Rubramin PC .5 mg/L

Rubramin PC 1 mg/L

Rubramin PC ~5 mg/L

*Control

c

c

6.30

c

c

6.40 ,:t.05

6.40

c c

6.40 6.45 ±-05

6~43 6.,50 ,:t.0346

c

c

6.43 ,:t.0346

6.40

~-~-----·----·------Ca 10 p 10

6 .. 40

Ca 20 p 25

6 .. 45

C = Compatible

Ca 25 p 20

6.'+0

Ca 15 p 40 6.55 .

Ca L~O p 15 6.30

Ca = Calcium Gluconate in mEq/L P = Potassium Phosphate in mEq/L

*pH of the Amino Acids/Dextrose Solution with Electrolytes

Table 10

Hlysical.Compatibility and Corresponding pH.o:f Aquamephyton with Various Concentrations

of Electrolytes Admixed to Equal Parts 8% Amino Acids Solution and DeA.'trose in Water 50%

Aquamephyton 10 mg/L

Aquamephyton 5 mg/L

c c c c~

Aquamephyton 10 mg/L

Aquamephyton 5 mg/L

*Control

6.35

6.35

Ca 10 p 10 6.40

c

6.45

6 • .42 ±·0574

Ca 20 p 25 6.45

6.45

6.45

Ca 25 p 20 6.40

c c J 6.48 :t,.0346

6.45

Ca 15 p 40 6.55

6.40

6.40

Ca 40 p 15

6.30

C :;; Compatible Ca = Calcium Gluconate in mEq/L P = Potassium Phosphate in mEq/I,

*pH of Amino Acids/Dextrose Solution·with Electrolytes

Table 11

Physical Compatibility and Corresponding pH of Solu B J!,orte 10 ml/L, Folvite ·5 mg/L, Rubramin

PC 1 mg/L, and Aquamephyton 10 mg/L with Various Concentrations of ElectrolyteEi Admixed to Equal

Parts 876 Amino Acids Solution and Dextrose in Water 5076

Sample No. 1

§~_P:l.~ NQ._ ? _ L c c

5.90

c c c

c c c

6.10 6.05

37

c ] c

5.98 Sample No. 1 6~25 +.0346

Sample No. 2 5.90 6.10 6~05 6.25 L_ Ca 10 Ca 20 Ca 25 Ca 15 p 10 p 25 p 20 p 40

*Control 6.40 6.45 6.'+0 6.55

Physical Compatibility and Corresponding pH of · MVI 10 ml/L, Fo1vite 5 mg/L, Rubramin PC 1 mg/L,

and Aquamephyton 10 mg/L with Various Concentrations of Electrolytes Admixed to Equal Parts 8% Amino

Acids Solution and Dextrose in Water 5096

Sample No. 1

Sample No. 2

Sample No. 1

c

c

c c

c

c

c

c

6.33 6.48 6.43 6.55 ~-0346 ~-0346 ~-0346

5.98 .:!.;..0346 --

Ca 40 p 15 6.30

c

c

6.40

Sample No. 2 6.33 6.48 6.43 6e58 6.40

*Control

.±.• 0346 .:!:.• 03L~6 .±.· 0346 .:£.• 0346

Ca 10 P ··1o 6.40.

Ca 20 p 25 6.45.

C = Compatible

Ca 25 p 20 6.40

Ca 15 p 40 6.55

Ca L~O p 15 6.30

Ca = Calcium Gluconate in mEq/L P = Potassium Phosphate in mEq/L

*pH of Ami.no Acids/Dextrose Solution with Electrolytes

Part 3. The apparent absence of physical '

incompatibility in Part 2, permitted utilization of these

mixture.s for examination after admixing Iletin .U-40 insulin

in increments of 10 units/L. The amino acids/dextrose solu

tion underwent prior addition of electrolytes and vitamins. ·

Observations and pH readings were performed in accordance.

with previous procedures. Table 12 reflects utilization of

Solu B Forte 10 ml/L, Folvite 5 mg/L, Rubramin PC 1 mg/L and

Aquamephyton 10 mg/L. MVI 10 ml/L, Folvite 5 mg/L, Rubramin

PC 1 mg/L, and Aquamephyton 10 mg/L were employed'in Table 13.

These admixtures failed to demonstrate physical

incompatibility over the time interval of study, except in

t.wo instances "'here numerous whit~ crystals appeared on the

surface of the solution at the end of the 24 hour period.

-f. r

~~ .

Table 12

Physical Compatibilit'Y of Iletin U-40 Added .to Various Concentrations o.f Electrolytes and

Vitamins Admixed t;o Equal Parts 8% Amino Acids Solution and Dextrose in Water 5096

Insulin 10 u/L 93 6.15 6.10 6.30 5.99

39

0244 ±·0479

Insulin 20 u/L ~8'--':"9-=-=------=6=.'-=Cl~4--=-:-_______ 6"-"".-"'0.~5~------=6'-"-.· 31 6. 01 H-~~-~==:......=c:.__:=-~--l-+.0479 ±•0264 _t.0264 .±,.0264

;·:

Insulin. 30 u/L

Insulin 40 u/L

Insulin 50 u/L

*Control

5.90 6.15 6.13 6.30 .±,.0282

6.01 ±-0264

5-93 6.19 6.11 6.36 5.98 .±,.0282 ±$0264 .:t-0264 .:t-0264 .±.-05

5.93 ;t.0282

Ca 10 p 10

5.90

6.18 6.13 6~36 +.0241+ _±.0282 .:t.026LJ-

Ca 20 p 25

6.10

Ca 25 p 20

6.05

Ga 15 p 40

6 .. 25

Ca = Calcium Gluconate in mEq/li

6.05

Ca 40 p 15

5 .. 98

P = Potassium Phosphate in mEq/L

*pH of Amino Acids/Dextrose/Electrolyte/Vitamin Solutions

Table 13

Physical Compatibility of Iletin U-40 Added to Various Concentrations of Electrolytes and

Vitamins Admixed to Equal Parts 8% Amino Acids Solut-ion and Dextrose in \Vater 50%

Insulin 10 u/L 6.24 6.43 6.40 6.55 .±.· 0264 ±.· 0282

40

6.35

Insulin 20 u/L 6.30 6. 5_5--------______,..6.__ .• _ 3 9'-----~1-~~ ;t.0264

Insulin 30 u/L

Insulin 40 u/L

Insulin 50 u/L

*Control

6.35 6.45

6.29 6.39 +.0412 ;t .. 0748

6.41 +0.1

6.43 ;t.0282

6.55

6.53a .±. .. 0282

6.33 .±.·0282

6.45 6.41 6.58b +.·0264 .±.~0264 ;t.0282

6.39 ±·0173

6.35

6.38 .±, .. 0282

~-----·--·------------Ca 10 p 10

6.33

Ca 20 p 25

6.48

Ca 25 p 20 6.43

Ca 15 p 40 6.55

Ca = Calcium Gluconate in mEq/L

Ca LW p 15 6.40

P = Potassium Phosphate in mEq/L

*pH of Amino Acids/Dextrose/Electrolyte/Vitamin Solutions

a - Precipitation in both samples at 24 hours

b - Precipitation in one sample at 24 hours

41

Part 4. The concluding study examined selected

antibiotic admixture to: a) the amino acids/dextrose solu

tion~ b) the amino acids solution in combination with "usual"

therapeutic concentrations of electrolytes, and c) the basic

stock solution with 11usual" therapeutic levels of electrolytes

and vitamins judged compatible from previous determinations.

The investigator felt that, in a clinical situation, these

mixtures should be in combination no longer than 12 hours.

This would include time of preparation to the completion of

infusion. Consequently, the visual observations and recording

of pH values were made at closer intervals.

The data found in Table 14 were gathered from the study

O! section (a).· The information presented in Table 15 inferred

a progressive laGk of compatibility of antibiotic ad.misture with

increasing levels of electrolytes. The Polycillin N exhibited

the greatest degree of instability in section (b) of this por

tion of the investigation. The results of the examination of

solutions described in section (c) appear in Tables 16 and 17.

Elevated levels of electrolytes seem to decrease readily the

stability of the solutions containing Polycillin N. The utiliza

tion of MVI appears to render these combinations incompatible

to a greater extent than Solu B Forte. Those solutions con

taining MVI and elevated levels o.f electrolytes demonstrated

lack of compatibility after eight hours. The precipitate

appeared as translucent crystals, which adhered tenaciously

to the sides and bottom of the glass flasks.

Antibiotics

Polycillin N

Polycillin N

Kant rex

Kef lin

Garamycin

Table 14

Physical Compatibility and Corresponding pHiof Antibiotics with a r1ixture of Equal Parts 8%

Amino Acids Solution and. Dextrose in Water 5;o%

Concentration --~ :·

pH of Admixture (Time ]:nterval in Hours)* o 1 2 4 1 6 8 24

500 mg/L 6.4-0 6.30 6.30 6.30 6.30 6~30 6.25 6.35 6.30 6.30 6.30 6.30 6.30 6.25

1 Gm/L 6.50 6.40 6.40 6.40 6.35 6.35 6.35 6.L~5 6.35 6.35 6.L~O 6.40 6,.35 6.35

500 mg/L 6.10 6ol0 6.10 6.10 6.10 6.00 6.10 6.05 6.05 6.05 6.05 6.05 6.05 6.05

2 Gm/L 6.20 6.20 6.20 6.20 6.20· 6.20 6.10 6.20 6.15 6.15 6.25 6.20 6.15 6.10

80 mg/L 6.15 6.20 6.20 6.20 6.20 6.20 6.20 6.20 6.20 6.20 6.15 6.20 6.20 6.20

Control Amino Acids/Dextrose Solution pH 6.15

*Visual obseJ."Vations failed to demonstrate physical evidence ]of' incompatibility

~

Antibiotic

Polycillin N 1 Gm/L

Kant rex 500 mg/L

Keflin 2 Gms/L

Garamycin 80 mg/L

Table-15

Physical Compatibility and pH of Selected Antibiotics Admixed to Various Concentx·atioil1s of Electrlolytes in a Nixture of Equal Parts r::IS% Amino Acids Solution and Dextrose in Water 5

10%

!_

pH Reading Time In:!; erval (hrs. ) ~isual Ex:amina·tion T me Interval (hrs.)

0 2 4 6 8 12 24 0 2 4 6 8 12 24

6.55 6.60 6.60 6.60 6.60 6.60 ppt c c c c c c X 6.55 6.60 6 .. 60 6.60 6.60 6.60 ppt c c c c c c X

6.35 6.35 6.35 6.35 6.35 6.35 6.30 gl c c c c c c 6~30 6.30 6.35 6 .. 30 6.30 6.30 6 .. 30 c c c c X X

6.50 6.50 6 .. 50" 6.50 6.50 6.50 6.50 gJ c c c c c c 6.50 6.50 6 .. 50 6.50 6.48 6.50 6.50 c c c c c c

6.50 6 .. 55 6.50 6.50 6.45 6.45 6.45 cl c c c c c c 6 .. 50 6.50 6.50 6.50 6.45 6.45 6.45 c c c c c c c

Electrolyte Content: potassium phosphate 20 mEq/L and calciuj~ gluconate 10 mEq/L I

Control Amino Acids/Dextrose/Electrolyte Solution pH 6.40

ppt = Precipitate C = Compatible X = Incompatible

-I=' \J.i

Table 15 (Cont.' d.) . .

I

Antibiotic pH Reading Time Interval (hrs.)· '[visual Examination

4 6 24 ~~ime Interval (hrs.) ·

0 2 8 12 ~ 2 4 6 8 12 24

Polycillin N 6.60 6.,60 ppt ppt ppt ppt ppt J X X X X X X ~ •I

1 Gm/L 6.60 6.60 ppt ppt ppt ppt ppt ,. X X X X X X

Kant rex 6.'+0 6.40 6.40 6.40 6.40 6.l~O 6.35 0 c c c c c c I

500 mg/L 6.40 6.40 6.40 6.40 6.40 6.40 6.35 G c c c c c c I

Kef lin 6.50 6.55 6.50 6.50 6.50 6.,50 6.50 0 c c c c c c 2. Gms/L I

6.50 6.55 6.55 6 .. 50 6.50 6.50 6.50 a c c c c c c I

Garamycin 6.45 6.55 6.50 6.50 6.50 6.50 6.50 q C C C C C X 80 mg/L 6.50 6.55 6.55 6.55 6.55 . 6.50 6.50 ci c c c c c x

Electrolyte Content: potassium phosphate 25 mEq/L and calci+ gluconate 20 mEq/L

Control Amino Acids/Dextrose/Electrolyte Solution pH 6.45


t

Table 15 (Cont .. ' d.)

!,

Antibiotic pH Reading Time Interval (hrs.) jvisual Examination

0 2 4 6 8 12 24 ~ime Interval (hrs.) · s. 2 4 6 8 12 24

Polycillin N 6.70 6.70 ppt ppt ppt ppt ppt ? X X X X X X 1 Gm/L 6 .. 70 6.70 ppt ppt ppt ppt ppt ? X X X X X X

Kant rex 6.50 6.45 6.45 6.50 6.50 6.50 ppt c c c c ? X X 500 mg/L 6.50 6.50 6 .. 50 6.50 6.50 6.50 ppt c c c c ? X X

Kef lin 6.62 6.62 6.62 6.62 6.62 6.60 ppt c c c c c c X 2 Gms/L 6.62 6.62 6.62 6.62 6.62 6.60 ppt c c c c c c X

Garamycin 6.60 6.60 6.60 6.60 6.60 6.55 ppt c c c c c X X 80 mg/L 6.62 6o62 6.62 6.62 6.62 . 6.60 ppt c c c c c X X

Electrolyte Content: potassium phosphate 40 mEq/L and calci+ gluconate 15 mEq/L

Control A1lino.Acids/Dextrose/Electrolyte Solution pH 6.55


~ \.J1

Table 16

Physical Compatibility and pH of Selected Antibiotids Admixed to Various Concentrations of Electrolytes and Soltl B Forte

to ml/L, Folvite 5 mg/L, Rubramin PC 1 mg/L in a Milxture of Equal Parts 8% Amino Acids Solution and DeA~rose iniWater 50%

____ .,. ... ___

,•,.

Antibiotic pH Readin~ Tim~ Interval Observation Time hrs.; Interval (hrs.)

1 4 8 24 1 4 8 24

Polycillin N 6.25 6.20 6 .. 20 6.15 c c c c 1 Gm/L 6.30 6.20 6.20 6.20 c c c c

Kant rex 6.05 5-95 6.05 6.00 I

c c c a 500 mg/L 6.00 5.,95 6.00 5-95 c c c c

Keflin 6.15 6.15 6 .. 15 6.10 I

c c c c 2 Gms/L 6.12 6.12 6 .. 12 6.10 c c c c

Garamycin 6.12 6.12 6.10 6.10 ·1 C C C C 80 mg/L 6.12 6.15 6.12 6.10 C C C C

Electrolyte Content: potassium phosphate- 20 mEq/L and ce.lc~iJm gluconate 10 mEq/L

Control Amino AcidslDextrose/Electrolyte/Vitamin Solution pH 16.40

~

Table 16 (Cont'd.)

A..'!'J.tibiotic pH ReadinZ Time Interval Observation Time hrs.) Interval (hrs.)

1 4 8 24 , 4 8 24 ......

Polycillin N ppt ppt ppt ppt X X X X 1 Gm/L ppt ppt ppt ppt X X X X

Kant rex 6.05 6.00 6.05 6.00 c c c c 500 mg/I~ 6.10 6.05 6.,10 6.00 c c c c

Kef lin 6.20 6.20 6.15 6.15 c c c c 2 Gms/L 6.20 6 .. 20 6.20 6.15 c c c c

Garamycin 6.15 6.15 6.15 6.10 c c c c 80 mg/L 6.15 6.20 6.,20 6.10 c c c c

Electrolyte Content: potassium phosphate 25 mEq/L and calciJm gluconate 20 mEq/L

Control Amino Acids/Dextrose/ElectrolJ~e/Vitamin Solution pHI6.45

.:!J

Table 16 (Cont'd.)

Antibiotic pH Readin~ Time Interval · Observation Time hrs.) Interval (hrs.)

1 4 8 24 1 4 8 24

Polycillin N ppt ppt ppt ppt X X X X 1 Gm/L ppt ppt ppt ppt X X X X

Kant rex 6 .. 20 6.20 6 .. 20 6.20 c c ? ? 500 mg/L 6.25 6 .. 20 6.25 6.20 c c ·? ?

Kef lin 6.35 6.35 6 .. 30 6.30 c c c c 2 Gms/L 6 .. 35 6.35 6 .. 32 6.30 c c c c

Garamycin 6.30 6.30 6 .. 30 6.25 c c c c 80 mg/L 6.35 6.40 6 .. 35 6.30 c c c .X

Electrolyte Content: potassium phosphate 40 mEq/L and calcij~ gluconate 15 mEq/L

Control Amino Acids/Dextrose/Electrolyte/Vitamin Solution pHI6.55


~ CP

Table l?

Physical Compatibility and pH o:r Selected Antibiotic 1s Admixed to Various Concentrations o:r Electrolytes and MVI Cohcentrate

5 ml/L, Folvite 5 mg/L, Rubramin PC 1 mg/L with a Mli.xture of' Equal Parts 8% Amino Acids Solution and Dextrose in ~~later 50%

-~ _.:__-:-..,;--_.

Antibiotic pH Reading Time Interval (hrs.)

1 4 8 24

Observation Time Interval (hrs.) 1 4 8 24

Polycillin N 1 Gm/L

Kant rex 500 mg/L

Keflin 2 Gms/L

Garamycin 80 mg/L

6. 1+5 6.45

6.35 6.35

6.45 6.45

6.40 6.45

6.35 6.35

6.40 6.40

6.40 . 6.40 6.40 6.40

6.35 6.35

6.35 6.35

6.35 6.35

6.45 6.45

6.30 6.30

6.35 6.35

6.35 6.35 6.35 6 •. 40

c c

c c

c c

c c

? c

c c

c c·

c c

? c

c c

c c

c c

X c

c c

c c

c c

Electrolyte Content: potassium phosphate 20 niEq/L and calciuJn gluconate 10 mEq/L

Control ·Amino Acids/DeJ<..-trose/Electrolyte/Vitamin Solution pH !5.40

..;:: ·-.o

Antibiotic

Polycillin N 1 Gm/L

Kant rex 500 mg/L

Keflin 2 Gms/L

Garamycin 80 mg/L

Table 17 (Cont:'d.)

pH Reading Time Interval (hrs.)

1 4 8 24

6.50 6.45 6.40 6.45 6. 50 6 .. 45 6.·4-0 6.45

6.40 6.40 6.40 6.30 6.40 6.40 6.35 6.35

6.45 6.LJ.o 6.40 6.40 6.L~5 6.42 6.40 6.40

6.45 6.40 6.40 6.40 6.45 6.40 6.40 6.40


c c

c c

c c c c

c ?

c c

c c

c c

c X

c c

X c

c X

X X

c c

X X

X X

Electrolyte Content: potassium phosphate 25 mEq/L and calciuJrn gluconat~ 20 mEq/L

Control A11ino Acids/Dextrose/Electrolyte/Vitamin Solution pH !5.45

\J1 0

Antibiotic

Polycillin N 1 Gm/L

Kant rex 500 mg/L

Kef lin 2 Gms/L

Garamycin 80 mg/L

Table 17 (Cont '.d.)

pH Readin~ Time Interval hrs.) .

1 4 8 24

6.60 6.55 ]?pi; ppt 6.60 6.55 ppt ppt

6.50 6.45 6.50 6.40 6.50 6.50 6.50 6.40

6.55 6.50 6.50 6.50 6.55 6.50 6.50 6.50

6.55 6o50 6.50 6.50 6.55 6.50 6.50 E?-50


c c

c c

c c

c c

X X

? c

c c

c c

X X X X

X X c X

X X X X

X X X X

-. . I I Electrolyte Content: potassium phosphate 40 mEq Land calcimn gluconate

Control Amino Acids/Dextrose/Electrolyte/Vitamin Solution pH J3. 55

15 mEq/L


.\J1 I-'

~-~~----~~-~---------- --- --- - - ·- -·

52

Chemical Com:patibility Analysis

p.V. Spectrosco:p;y.. The previously described techniques

were employed to obtain standard U. V. spectrograms for the.~

solution of amino acids and additives investigated. Triplicate

·samples were diluted with double distilled water at the time of

scanning.,

A 5% dilution of the amino acids/dextrose solution

-n--~~~--n.pr-oduc~e-a-a-sp-e-ctrogram-w:i:th-a-A oi' 279-nm-\vi-t-h-second:a~L-Y"-----~~~~-max

~max of.267.5 and 287.5 nm (Fig. 1). A Beer's la\•1 plot con-

firmed the dilution best suited for the succeeding analysis

(Graph 1).

The examination of Solu B Forte, 10 ml/IJ, produced a

spectrogram. e.x:hibiting a A of 262 nm when the mixture was max diluted to 176 (:B'ig. 2). Similarly, MVI 10 ml/L, produced a

~ of 265 lli~ for a 2% dilution (Fig. 3). max ·

The application of U. V. spectroscopy become limit eel

after determining the concentrations necessary for obtaining

standard spectrograms for the remaining vitamin additives.

Obtaining a u .. v. spectrogram for Folvite necessitated a mini

mum concentration of 5 ,Ag/ml yielding a Amax of 282 nm (I!'ig. 4-) ..

Similarly, R.ubramin PC required. no less than 5)1g/ml, exhibit

ing a }\max of 257 run with secondary peaks at 27'7, 295, 305,

and 320 nm (Fig. 5).. In like manner, a A . of 21+1 nm with max secondary Amax at 262.5 and 270 nm was obtained for

Aquameph;yton, 10 fig/ml (Fig. 6). ~1hese data inc1icated either

the eoncentration of the vitamins must in increased for com-

parative dilution, or the undiluted vitamin/amino/acids/dextrose

solution would have to be scanned. !I'he former situation

would necessitate admixture concentrations·f'ar in excess

of'.those utilized clinically. Scam1ing of undiluted solu-

53

·tions raised questions concerning the interference of t:he

amino acids upon the spectrograms obtained f'oi' each vitamin~

r£1 0 5 ..... p:)

~ r./2 {:q <l!

2~- , i" 11 I i~i?~-?~ .I; : 11 i • ·1'\1)20

!\ , . . .. 'l.si , I '''' '-

'

! \' . i : : : 'I ! I ! i ; I : i I : i I I II i i Ill . . I . ' ' . ' I I .. i ~~ I I I ~--'-~

I' \ ; ; II:!., i! i; ; !': i '!II ! I: i I 'ill! ! . > I' , • · , '!: I · 'til : 1 :---t-t--f---i'f---j-1.1 . . .

1:' :.l;.:. T!l;_; .. :i;! j1l! i: : i\: i I il!

1 ......... I' .... ''' :''' . I II itlj,

i-:-+-!f-7--f---L1 ~'-J-j o. I ' ' : I ' i : i .

. :qJ::. :x~:~:;:; ;!-il::i i1!i ilil 1

l::·ft:~~ 1 ·:::::·:-:·~,' ::·:--~+;-:'~ -('~· :1-r r-m,_i

1 "''I" I"'"'. fi"·' ''I

~-~W-1 Li:, :Ul!L- ~,_fUiJ 1. ;: 'i. : ::. ~H:·:. :::: :::: r··:-·, ......... ! ....... ~~----' ------ .......... r '-ri_J ,:::(! :::i : \;;:: ::·:1!!!- ::,u;!.:l r:1i: 0r7!.:;:. :n: i; :::~t!

1,: i-1,

1----:-- !,-:---· .. j .......... , _____ , .......... 1 ..... _ .... , .. , .. .1., ....... , __ ._ I

' : : j . : : : . : . . . I : ; : : I ~ : . ; I : : I : I ! l LL.J..-' _. _!_: ~~.:J.:..:_~_-_: ~; :_·. :Ji_i_UJ I ! ::. ·:; -: ~· : : j . ! : . J'/.A, , : : : . n· ; , T ! ; 'l i: . :: . . . : : ..... -. ::\I ': : :I . :I '! :; r.··---rr-----,-----~-~--~-----\1-·---- ""''~!-'""' I : : .. :l .. ; : . : : : l : / . . ~ ; : \ : ; ; I : ; ~ ~ I \ ! ~ '' .. ,, . . . . .. I I'' . '.: 1\:::: i .. '. i '''I ~~:~: j: .. :·;·:: ::-~~:::: .:::1 ~-~-L~~L ~-; L~.:r:: ·-~ ~~~ :~-~:~ ~\~:.1-~L~~r~:!; ~, I .• : . : ~: . ! : I : : J! ! ~ : . I ' ~l· ! _: '\' ' I ; 1 ! ! ; I i : ·_ t. =: :\ : ::1: :1·';:::;: 1: ::: 11,\ n ! :: 1 ::! 1--: 1 I c--;~-:---1--- ~---·j···-'---;--r--~--P····-'·-'-~-'--1:: '·1\ ••. I :t: i;; i: <:: !] ] \:I: li i 1!! i '! -~-----l..,.--~-----.1_, ........ _;-r~---~-1 1::: l.\ !/:::1:: .. ,1 :;:j:;:\,,iilj'l:l ~::-:-:-+-:-~,....r---:: :r::~-r~~::·m~-i ~: ::1~~

l = i : : : : ~: -~ ~: : : · ; • ; --r~t~t· ! }~ · r : ! I ; i : ~ , t ... ;.. . w--·----·-r~-----~----- -"... .... L-"--_ --:--L-. .;....1 I ' ' . : : . ' : : : : : : : 1 : 1 i.: I\ : ' ; : : : 1 · I , ,, I· . I' ·~-4l2.r\' .. l''i I

L! •• i --H: h- r ,,i :u#~·l8t~J~J1 " " I . " t'' '- f • ~ •' _" ~ 0~ ~ ...L;_ll

Fig. 1. u.v. Spectrogram of 8% Amino Acids Solution and Dextrose in Water 50% ( 1 ·1;'-......

\'lavelength

>-max 279

,..... a

()'\ ('-. (\I '-'

r:£1 0 z ~ !:Q •:l::; b U) r.:q <!!

0 7~' I IiI' I'''.. ' ' ' i . • __ , - -I • -t· . -1 t .. --! . 1 • ·I- ! - !·· . - . ;-:-·· - l ·- . -~. ~--. -- ; .. •. r--:

+~;~~H4iH~f·i·:!: ~" .. :-F:Jt HJ o.6 ... :J., ---~-~·-:·r"-!--1-~- .... ; :----· -: H- :-··--.----i t-1

I I I :~.! ·I· ''I :_I I' I : I ' l ; ; ... i ·' '·' ·: ' . c _:_1 • • t ! 1 ~ • 1 • • ~ , r : l • • 1-1-- !----1 --1 -f -:- ·-T--l- ·r-- 1--· ~--' --1-- · --· -1---·- :-"T""T -:..-;---r--:-· I ' I ' ' ' I I I ' ' ! ' . . j I '-I I I . '1, ·I -' . 'I 'I' ' ... -' . ,.. ' . . . -. '. ~ . ---: ., .. I ;.. ' ' ·I ' I I . . : ·' i .. ' I .. i o 51. I .. r -~--I .. I ... [ .. ,.. _ --~-- , .I ... ,_,_, ___ j_ .J --1-+-- --i--!- _,. . ~--.

• I ' ' I I ' ' I . I I ' ·' l : I ' I ' .. : : ;

F~:f·i~ I~! ~-1--:L'I=:!~l-- r''l ~ :-·! . ·!~ .:\.~~:fFFf'!~]=l:-~·t-rJ () 4-~--·-T i"' ...... l_. ____ ,_._r! ..... 1 .. -j--'- t,._; __ , ___ ~--. ____ ;._ __ :

~. ~. i I i I J I : ! : i fi:::_l i:LiJLI+H:H-~f1 ~t·.tiJ

Concentrati ;n (196 o:f Full Strength)

Standard. Solution r, . "! ) \-•-

Graph 1

Cur1

·ve .for 8% Amino Acid;:::, and. Dextrose in Water 50%

\.Ti ..;::-

~"'t1 0

~ ~ g3 <!

22~ I . r=· ::df*ffi' ' '-~- J..foiJ~O Wavelength I· j:l '·.i ,.\, ll:i; 1\

--!-'-~'-- -r---' ' -~' ' '1 5 !..l.. . I ' I' ' ' 'I ' II --r--+---.J,- -~ r~~~ ~~-:~-.: ,~~: :_~H ~ H-:-~, . I . i ; " ' ' " ' I I I i I

-~---~-----~--- ~--- ~~--~-:_j · ~.~l.~-,..l-!-. I .. :. I .. j:. 'I' .. : ' I'. : i . I . i . I : . . ; ! '' : i:: ~!----+-:-·1---~-:-~--: 1----f. I ic-r.:..~

I ': . I I I . . I :·,:·-·!· ·: r::~\ 1-:l: J. l_t -. tJ---'--1-- -~---~-.1.-.9--i-o----. . I I ' I'\ I ., •j. · ... Ill ;--1--~~~ ~ . ··I-I'!. \--1 ~ .-. ~t~ '-: .; ~: :.1: l-l- ~----+-~--~. - _l;._:_ • --'-.s-f-Lt-\ : l . .: ! .! . : ; \ . I jl . : I ; ; i I ! ~\I _: __ :+ . ~-1-f-. ··-- r~-· -\ : _:_· ·f-c'---" -:7, .• i-~-1 . I . . I' . . i . . ; -I ... j . I ' I

;_~---· -' · ;__ __ r---·•..!.---~ .:._:_Li- -: ..... ~ -1-'- -il-

; 1 i r ! \ t> ·.··• ii :-[\" -t--tt-~:_:__t~-t7·&~1' it

l ' I I I . . ' I . j ' ' ' . I '

~.:-. _L_\ :-_r: :z_t' .~j_.J~~ t~ __ iH r: ~:_lr~~L h-.1-. ~ ~. i I . . ! I\ : .. I I ' : ; . I \ . ,· . . i. . I\ . I; . ; : I! ; .. I ____ ,_, ' . . . I • . I .: . ' ... .. . . ' 1"

J 'f---- - f 'ic itr. ~:~ : ! 1· .. : ... 1 · · • • : ~ • t , • • • I • ·•· .; :. • • :. : • : I ,

. ' p- . T:':,. \·:i'T; IIi J~I:l J iiJl~--~ :~'1; It _ r · ____ j : : f>i ~ > 1 . · : .\: _: 1 ~ : :_ u

I . ! J : . l .. :;\ < :: t-~:- ~~~ .-~-,--·t'-~ ~--~~1::-1: . ; I.. . I: ., .. r. ·,

li~~ - . .- l. ;·-. • .. : :·; ;-- . .:..., ·--=r7-l 'j.. • • I I'' ll .. i l I I j II I. . - '! . t • :I.. , l.O'

A max 262.5

Fig. 2. U.V. Spectrogram o.f So1u B Forte (0.1 ;<1/m1)

.......... s L."'\ •

(\j w C\J

'-..-"

Pit 0 z ~ ~ 0 w !XI <I!

• i I • I ' ! . ! I : ' j I i : I I : •. : I i

o.JT-1jj ir!l i! i '!: i' :~2:-~~; .J~t]J~! !-- r-- . · i i ·' --1- r 1 ·i · · , jj ; l : :-- i : · t ·1 ;--! ·-:- r ·: -~ - r-· t-·;:'-r

- i ~---! --;- ., .... ! ., .. ; · - 1 1·· T ., · · ! · :-· ; -:--: :·-1-...,r--:- -- ; ,-- 1'- ·-:t

' . . I . \. ., "; I . ! .. . . ' I : .. : . ! ... --·-; ! ~ I .. - __ , -:-:-r. , •· • ' 1 I : I ! ! ; 1 1 1 I ! ' 1 .:.J o.Ltlt -~-' ..

1. -+-~ .. _

1 . , ___ j _ _, _1 __ _, ____ .... -~----:- , _ _, ___ ;_ __ _

~~: l~~----l--1-~ ~---; .... Lr.::;~--~1 -·l::t;.:L~-J:~-'f..: -1:~~-..W -~L_; I, 'I ·I I .j. I I I i ! ·I ' [ . I I ... . : ,. : .: !. i ' ·i .. I . -! . ..;.: . 1- I I I . I ' i ' I : I ' ':.:::.:J ,; ' . . . ' .. , I 0 • 2 !.- L! -+ ,--::-,L+~~ T. c -:-- , -Tr---1• -.c t ~--; __ : _· 1"1~n-;--~~i~-~JI f

! ' i : I 1·' I : li I ~ I I I : ' ' I ! ! : ' .. '· i ·-!· . i--·· -+:+- ·--· -+- -1··-t·-r- .. '· -~---:--1--:- ~--.- ·--~c--l-t- ·•· -+ .. r-L .-;:-:--1 ' I I j t· I .IJ ' I ' I ' . ' I • ' .. I·: .:t · ., "" 1· I ,.,. ·- ·. ·+d .... --[1-- ,.· r::i :::r:'·'~ -• -!----:"I T-t , .. '-t--"1--:--+

L.L~--~-~ ...:.. _: i:.l±L.l.JJ.,jL:t..:ili..±diJ .... b .. •::: :~:...:L.£~,;-:'1 .= • :q,

0.5j 1.0 1.5 Concentration {1% of Full Strength)

I C}raph 2

Standard Curvel.rcr Solu B Forte

... ;

\Jl \Jl

r:il 0

~ ~

~ CQ P=l <::::!

~~~~ ,, ' ' ' 9'7() .i " ,,··~-~ ., !' . . ' I' . . I'''' . I. I' I I I' ! . : ' I I' ! ~ ' I ! I ' . i ' : ' I I I I ! I ! i ·1.5' i 'L

: .,, ..... ,,I'·" "'I I . ' . I ; • • • : : i L • .; i I I 1 I ! •

~ ' I ' ; : l ! : : ' ! : ' ' ; --t 2- .L..i.~

l , ,. ' I 1 '• , .. · : • 1 , .. , . i 'I' . ''II• I.,, 1111 '• ·,Ill'' ' I '' ' ' I I ' 'I '

I' ''' { ' I' ' I' I '' I 'I'' ''.I \ • ' I' I j! 1 I: L t

--~ --r,----,..-- ~·- ~ . 1.1- -·-·-

. ; : . l : . . . I . ' . ' j· . ' " : ' I ; I i ' ' i ·: ; . ' '. !.•: JJ•,:•

Ifill ~--~~--~-·- ---~-~ j-l.Q ._:_~

! . : · I . ; ·1 :: I· . I . . .. I' I '' I'. :til

~- :.-~L....-.-1~--...:..t.:._.:..:. ~-L 1.~.:.- f ___ ,_t:~~.! • ' . . . . I . ' . • . • 1 j : : I i . ' I

hl.i · ,. :: ·

1-.·-J .. 'Ti· •:tmi. il;;

-~:: ~ : • •

1

, • • • I • ·': \,1: :' ~~; :: j, .. ,.... 1 .. I· .. l,f '•I' I'' .... ,:,! : : ! . : : . : . ' . . ""' , : . . . I : ; , . 1 ~ : ; ! : i ! ---~---- ·---~·~-' ----L~- _,_II~__ -----~--,i-

1 .. · ~~- ' · 1 ' 1 ' " '!:1 I I ' ' . ... l .... I ... ,,.,1.· . , ,.,~

< ; I ; ~: . : · {·.: : . ; . ' ; 1 : : : ; : it-i i : . :-~: : ; i ! t .. ·---.·t-··--- ----~~---- ·--··w--- ,_, ___ .,.1L -------'·"1'·. :: y!: .. -,·1·: \: '::I i:!! I: : 'I!! l 'I

! i I . i ; • . ' ' ' ' i " i ! I j • " '

r --- --- --- -- . ......,, -r-r--1 '. ' 'I I ' 'I ' .. ' { ·:.j' I' . ·!:': '.

: ; . I · . '/ i . · l , . : . : : : : l ; : : : : ! •

t-.. - I - - • - • • - t-·- "I' t ... j .. -- -" ·-·- -l .... ·----.-1·"-· •-i· . : : . ! : : ./ i : . : l · · : ! · : I : : ; i :.:1.·:· . I . \·I·': ·:·]'1!· ·!:':' r-:--:-7.:..1-- - t ___.___ __. __ ,__ -- -- - .G --r~-·-·. . :' ·1 '. ' ., :' ' : . . . ::: '. " ..

' I ' I ' • I ' • • ' • ~ l : I I ! ' I I • ' I I' i -~ :. t: ~ _. ,. . --~-'- : ., : . 1. 1 : : ! , .:.- .:.:J.. _.:_ __ j.; _,_, j . : ; . i ! . I . . : \ l ~ : : ! : I : : i t I : ~ :

. '.'I''. . . I·.' I ' ' ' ,.I 'i;

. . . L-: -r--:-:-r.:..:....:.. -: . : ~-. : -·-.s.:..,-:-,~+-1 .. ····' ... "''l"' :I'"' . . . : · . H~-- ~~ Al~~-~~ ~; :+ 11+- · -~-r~~-~

.. , .. / 'I ... 'I''" '"I!•' I' :1 ... I · • · · · • • · 1' • ·' •. :'' _4

_......:.....:.._1 . ·. · '· A · I: : :: . 1. · · : :; i ': ' ·I:':; I If •••• ; . • • . ! . ~ . : . ~ : I i : ! i I . . I • i

1-: . : ! I : :-: ·J:-;: : ; ; ~ !\' j j ~ : ; i i ·[·!.1. -: ; I ! i ; ! ___J_JC ... , ...... ···!\,.,,.,, .. ,I .... ~. ~~~ l~ : : • !· : . . ; . i \J .: : ; i > ; i ! .3 I i : ... . ~;-;--:. ~-;. . .. -~ ;-·:-. --·--;-: i. i: ~ --: i-:- t d-·- -- -· ! -t".

J-fui:~_L __ ~I: .:: :~:!;!; --zlLJ

l· ... ~; ,,, ' ·. :,::;l T : • • : i . : \; I. ; I ' : • ' I , : ; :

""1-:"!' .. j--- .. 1· "1 .... ':·:~ 1 ::· ... :h·::;j

¥0=~~1;=;1JrFii Fig. 3 .. U.V. Spectrogram of IVIVI (0 .. 2 _;tl/ml) .

Wavelength

An ax 265

,-..

E tf\ \._() C'J '-/

t:=.l 0

~ r:q

~ (/) r:q <:

---~

. ' I I ' I I .. I • { I I . .. -; l ., I . -~· I II . I ! l -I i ' . . I '. ' • - ... - .. : -! •

.:/ : • ' 1 ! : l ; . I • 1 I : ; I l • • . ·;----·,- L~--,-'~·--· ,--~-:- ; -: I·; j ... - j- ; ·; -r-·---i ·-! -·-·;-·-J

' I . t j ~ I I . I • • ! . J • 1 ~ :' I I -··-~ I • I I •• I I • I l . . . . I I ~--'--~-- 1

...

1

. 1.... ._

1

__ ...

1

. _. I., , ;-- .. , ... --: _____ j_ __ r' ...... ____ , __ _, 0 8 I I l . ' I I ' I ' I ' ' : .J • !----,--·1 , ... "i" ,., 'i.;. :· :-·i j. ·t· -:-·-;-· i

.,' -~~-·~,- .. ] . .,,·- .. , l-1 ! ·r !. i,·- :· i ·:--!"~~~ ~-- i-~-;,:-~---·

1

:_ f"'---L-1.- ... , ... ,: .. 1-l -t-· I_, ! '---1- -:--: .. ;_:_ .. j_' .. t---;--'-i-' j 06 l r 1 1 ·It':·',.,., , .... -1

.. .. -1 I .. t • • • 1 .; j · ·1· • ·I • · ... · t· · 1 • • ... ·; c·-,.,{ • ! I I ' I I I I .. I : : I • I : • t I i I' ...... j .. l +·j· L .. l.. --I· ... , .. --1- :-+ !" ·----· ~ ... t--- :-··-·--'·] I . I I ' I ' i ~ ! I I ' ' ' . : ' ! • I ' ' : ' I ' I ! I . , I . I , . ' . ' .. , ! . ' . r·· '_,, =- •

l I I • ' •• j • I I • I : ! I .• : ! : •

0 LJ.'--1..,.. ~-:I'·'' ~-,-t--,--t-· :--'-i--··!·· :-;---;-- ·:---··--'--j • I· I I i ' i l I i ' I ' I : I . ! . : 1 ·: I. . ,. ; - i ... -..: ' ' ' ' I ' . I I I {

(~-i-' ~--; ,. r.:· .. -;--· ·t:-l: (::·.r --:-.! .: t .(-t-;~-r-~-~-~~r=-=1

0 21·--·--1- -- ;-- . j -[- , __ , ___ ., .. : .. ;. , .... , ..... J ..... ; .... " -1----•--:· ----·-·: ___ j I I ' ' I ~ I I ' I : I . I I I : l" I { . ' . 4 • •

1 I' l I I : .. : • : • ! ' : I i· .... ! :. ! ·-·. i I l , i ' . I ; I : ,. I • : : I . I .. l !:'-, -- -~-LI -; .. ! --,--.--:-.I l --+-,.--'1-·--'-t· ·---·r· -~-r-,.--'__._·r-- ""-7--+-'-;• I : . I' . 't''. 'I I. I.: ... •.f :· ·l·""j--·J-~ :-- j--;-T-! -r:i-:r-T 1-'-- .·-!- 7-"f'~-i-·: •-·,-T·:-ic;"''l · -··•-~l--·- 1-t..-J. __ !_:...1, __ :_~-d~ LJ-~J __ !,_ __L ___ Li_-~~

1.o I 2.o 3.0

Concentratidn (% of Full Strength)

I Graph 3 Standard CuJJ~ve for NVI . \J1

0'1

J:il 0

~ p:: 0 w lXl <I!

f!~? · t' · • : • ' : - , · • ~ t.ra 1

f'. · " , • ' , 1 • ' ' • " ve ength ( )

: ' t+-88' ' ! I i : I : : i . " I . nm · F:l--:'"" .. n"'"':t""T"',.._...----1 ,.- . . . .. ~-1_. ' I ' ' I ' ' ' ' I . ' .... ' .. • -jl . l:: .:.~:,,;- :1 ,•. ·.: ,•I!· ,,, !I! ~.o ··r'--1--:- ~-~--l_L .. l .. H-----1.-l :_l_tT-:! ';. -

' . ; II I . : .. ' ' ' . ' ' I I : I : m· ··- I ' ' I I . 1 -L- ·- : I r . j I ... I I 1 :1..:_:_ .. : ·

1

· ! 1 • ! I : , 1

1

· 1 ,

1

• : 1 • , 1 1 1 .~.. · , ... 1 : . ,. 1. ;_ c :u-:-·,-- ;---(--;- +- :. :, _ _:_ __ ; - ; -~- -- , j- 1 . ~----~~ c4.,, I, I ! 'II I I; 'j":"C·-t ., .. ,:_ .. LL L[.,. I'' · -'·! i. :. , ~ T~- ·c+-'cj .:-!·~--r.': _.:: ~-,,,,:-r-.:..;--u-1· o 9-TI. ·J·~..._,_, ___ ,_, !1-: :-~-~-,-J--:- .. : _____ :. :·-- :·· ,.,_! _ _.

:::1:1. : l> y: i( ijill'l: • ~(i•rt'·+,~-j-:.;_·::.: ... ;~~- .I::.--:-:-,:-: !-~----+-:...1 .. _:_. ~ L~..: I ~l~ : ; : : l ' • ' ' '..._._, .:.:. ! ! ! i ! ,I I T : ,+ L +i '·I : ! 'T"- i "'f:.-:t• ,. •.. ' ' . ._.;

>r: :"r:· ~c( .. .. :"I'Lli '''1'1'· o.s-i--'-r:~•· d' ,JLb · ·r~·· -·~-:-.• .. : l , :lf I,, I ' , ••• ; ' ' ·.~ Til' rt: r ! ! if::ilJ i !·~+~~ .i ·+c'. _I il -~ - . J I . " . . .. iTIT ,.~+ . . ' . I r I . I , -. --~ ---- --·- - , • , . . -- - I . ' :::: :i::l;-:- ..

1

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o; _ :! :·.- .. ·:1·.· :· ·!.· 1 ·-- -;.,-,---:.t- 11~ I ~ ~1~ter Jr; ' · · '-" · · ' ' : ' 1 ' ' • ' : : 1 I 'JlJ ' Graph 4

_g._4. U.V. S ~ St Fo1v~te (~ g/p le)ctrogram of andard 9urve fo li' . / rr m r - olVJ.te \.11

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Fig. 5. U.V. Spectrogram of Rubramin PC (15 .lfg/ml)

Wavelength (nm) :T'i">':'IT --:)··· :-r-r-::-· . , . r-~-~-:·-r-.--r--;--, -, -:-;~

~n~~ 257 277 295 305 320

-.. a t'L'\ C\! '-"

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I I I . ' .

5 1 10 15 Microgriams/r·1i 11i1i t er

Graph 5 Standard CU.rve for Rubramin PC

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I · .• i

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~. ~- L~-~-;-~ 1 ::~- ;::. :,:: i:!: : .. jLU '+tc~- • -Y--"-:· :_~ J_, L;_f ;j +1 l . : . . . I : : . : : : ' i l i i . I 1 ! . l.. , .. ,. , .. f_;.. .. . I :- !... . ·~.. . l .J::j l. i . j . ,, . . ~w~ +·: 1 :·r; · : ~ ·: r:·r: iFf ; r·-f .. J.. .... L --tr~:~ -H ~ :-t·ttt-t=·-; . ; . -t=rr:;(- ... -+ 1 .. ; : . : \ . ' : I i l : : ·_ ; : . ! ! ; .• ' 0 .1 ' ! ! I I ,.. . 1 ... , .. i I . [ i I il 1_ .' .. I. .. : ·-'- .L -~J... 1... ,- : _L] 'E'' . . .. .... R+ - ' . I . ' ., :.j .. - ..... -·+ .. 1""1' , ... ·! I . 1 : I I ' I !j ~ I

3

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h-..:~-1

~-H·:·. _:'t_;:1 i:;~ ;::]:::. ,!i!Ji! 11i fvlicrograms/Milliliter

! . . . . : j : : . . ' .. ' ' ' • ' : ; . : J; i ' : : : . I f: f· I :.. · .. ·-: ·' · .. : · '::: · :jl; '· ' · · 1 ' i i Graph 6 l . ! . . : I : : : : ' ll.~. ; ~ :.: . .;: : : I :

f-:-:~[- · ·' I:( '..J:i:~ c-.:.: V: ·. :;iifq++ _. Standard Cu,rve for Aquamephyton 1:·:;·:1::·:··~-~::::: :::::;;:;· ::!: ;ii;llJJJJ L ~ t 1 ' . : 0~! : i . • I •• ' ' ! • -

FigG 6. U.V. Spectrogram of Aquamephyton (10 ,}(g/ml)

\Jl •.0

60

Having determined the standa:I.'d u.v. spectra for most

additives, various admixtures were examined.

Electrolytes. Potassium phosphate 25 mEq/L and calcium

gluconate 20 mEq/L were admixed to the amino acids/dextrose

·solution. As these agents precipitated at this concentration

in distilled water, only single additions were placed in the

reference solution. Duplicate scanning of each sample solution

alteration of the U.V. spectrogram·for the solution of amino

acids and dextrose compared to the standard.

Vitamins. The combination of Solu B Forte, 10 ml/L,

~d the basic solution of amino a~ids and dextrose produced

spectrograms indicative of possible chemical interaction. One

hour after admi:x:ture, scanning of triplicate samples produced

a u.v. spectrogram for the amino acids/dextrose solution

exhibiting two additional absorption peaks at 230 and 262.5 nm

and a hyperchromic shift in absorbance at 282 nm. The secondary

peaks normally demonstrated at 267.5 and 287.5 nm were unde

tectable. The U.V. spectra for Solu B Forte also showed a

hyperchromic shift in absorbance (Fig. 7).,

Four hours after admixture, the U.V. spectrograms con

tinued to modulate. The first peak seen at 230 nm exhibited a

hypochromic shift to 227.5 nm. The second peak disappeared.

and the major peak of absorption, initially 282 nm, shifted to

279 nm with a hypochromic shift in absorbance (Fig. 8)~ The

spectrogram for Solu B Forte appeared unchanged., though a slight

61

hypochromic shift in absorbance was noted.

The u.v. spectra obtained 8 and 24 hours after admixture

were essentially identical, and exhibited the most profound

alteration compared to earlier results. The spectrogram for the

amino acids/dextrose solutions displayed absorbance at 230 nm,

a second peak at 253 nm, and a significant hyperchromic shift

in absorbance at 280 nm. Similarly, the spectrogram for Solu B

Forte featured a slight bathochromic shift to 26?.5 nm. This

development was accompanied with a tremendous hypochromic shift

in absorbance (Fig. 9). This observation \vould tend to coincide

with a noted drop in concentration of those components which

initially influenced absorbance at 262 nm in the standard U.V.

spectrogram for Solu B Forte (Fig. 2).

These data appeared to suggest the occurrence of a

chemical interaction, progressive in nature, over the period

of investigation.

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l : I:: ~ :: ; > ~ i : t ; : i 'l i ~.___-__ ./

Havelength · (nm).

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-\nax 265

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fi!!~:' ••t ,1•+~r+,: m il '1~:~1[:~ lr,:,wir ~-i-m :;:r! )\-'HiP /:"j:IJ

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'' ' ' •jl' , , I ' ' .. I:. : ' I' i, ill . , . 'J'.. . . o-·' -· _1 'l:; l i I! f-!! i l; i I:·~· :. ' '. .

Fig. 7. U. V. Spectrograms o:f Equal-/Parts 8% A.'Tiino Acids Slolution and Dextrose in 'Hater 50?S (left) <:Lt!.d. Solu B Forte (right) One Hour Post - admixture.

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vlavelen§;th (nm) 2')" ' . . ~: It:; !j!l - 1 -'- --~·-

mo

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r_,, __ !_. _____ ~ _____ ! ____ L: ___ :_rFt_._~J!il'l~-~-:

I I I , 1 ' ' , ' I'! •

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r·:j;:~~ ·:·~:!T'_':-~::(r·:~·: :-::'!'i. !/1': rtll

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: ·:: ! .. "· i,.. :: 'j · . ::; 'i j i 1JT1j1 ~~~~·I ~ : : I .. • : ~ ! I : ~ : . ! • ' : I : : I I : ' • I ~--· ~- -~-· · --- <---"-· ..... \ .. -" .... > L i · l I -~ 'Jj-1 . . ::.I .. : l :I ! . . '\' :'. i: ' :. '! ! I ...... ', ... ~- •'· ' '1' 'UI I ~J ·-' l...L.:...!_. · ~ • .:_ I I _!_· ~ ..!J. _: --~ ... , ......... ,. l'lrl.i'

I ' • , ... ' --1 I ' ' ' . ' IJI' ' '.. ' .. ' - ' I ' ' ' " I I I ' • • • • ' ' j l. • • I .•. ' I ' ! --~--!-~- ----- ~-+---- --- t·----\ --:---- -t-.,-- ,..,. I, ,.... 1 ... .. . . . . . . .. "I. . , . I.. . I r~,. I', - :_I! '' '.: '!.: ' '' . J '::. i I:' iIi Ill! i'

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E-·T-- .. ~. ;~ :-:-· ~- : ·,~~-;. -: ;.~-~~~~:

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.. ..

)..max 227.5 279

Fig. 8. U.V8 Spectrograms of Equal Parts 8% in Water 50% (left) and Solu B Forte (right)

~ax 263

.

-~.o~ , I r • 11'~·1. f i , , : : ! I ·. :_ !-1 ,11 •1

. I. ·:~___,_,_ ____ w 15 . ---,. r- · · • · ' 1, · r.-~, : ... , 1 ! . , , : , . .

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~-,--!:L +/U-:- ·, ·!:-~:-t-::-1::-:: ' I' :I ; i . . : j: :. I .. , ' ·. : .:..! . ' .. :.J_--· \~;~;y_~ -~ i+crc . '. ~~~ • : . . ·';; I'' • 'I J i I;; ii \ i! l'' ;c.cLc

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c!; fil :s·;t .:· i::__ ~ f.,, ~~ t . r : . . i •• - I . _; : - _, ... - I - :- : : t1:; :;1· i:'iil ::. :i!lL>d j i ! ~ ; ! t r : I · : ' ! · ' · ~ ' '

Amino Acids So~ut.ion and Dextrose Four Hours Post! - admixture.

(j\ ~

pq 0 ~ !Xi 0::{ 0 w ~ <!!

\'lavelength (run)

>max 230 253 280

\nax 267.5

1 r;: • 1·1 J' ~-i 1 •' I 1. 'i; 1' I·: i •I , . j •

~ .:rJr~n= :!~::·1~,•~•·:f-::~ :!II II•, ,{nTTI. ,,,,l.:,.fr!i·l . :

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:: . I ! : : : : !!!: ! ~ :_! . ! ; : I : 1 ! . : ; : : i : ' ~ ~

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.:·.·~·::..:....:._~.:..-'': ... ;;:; :!:.•:: 2; . \I . ·j· . •I • ' i . . . I •. ' ... I

: : ' I I ; . . : : . ; :~ . ' . ' ~ . : \ . . . . ; ~ t - .. : : . . " - -[-........ , .. -,.. . . ·--- '. \- ..... ,_. t·· -· ,_ . ~ \· -~--·· ., .. 1 ..... I . : ; ''\. . ' ' 1 I ' ; · · ' · ' ' · -1 · · . ; · · · . .

m,~T ~H%~ trt+~ ~::~~-·~ Fig. 9.. lJ.V .. Spectrograms o:f Equal Parts 8% in \vater 50?{; (left) and Solu B Forte (right)

I . o

Amino Acids Ej·olution an~ J?extrose 8 and 24 Hourrs Post - aa.m1xture.

. ~;:.; .; .

0'\ +="

' i

i

65

In like manner, triplicate samples of MVI, 10 ml/L:,

and the amino acids/dextrose solution were scanned at 1,. 4,

s, and 24 hours after admixture. The amino acids solutions

spectra revealed immediate alteration vJith absorption peaks

at 272 and 275 nm (Fig •. 10) one hour after admixture. The ·

shoulder at 287.5 nm appeared to be retained. The spectro

gram for MVI lacked any apparent change.

After L~ hours, the mixture disclosed further progres-

sive deviation. A Am of 267.5 nm appeared with a succeeding ax peak at· 277.5 nm (Fig. 11) in the amino acids/dextrose solution

spectrogram. Concurrently, a noticeable hypochromic shift vms

noted on the MVI spectrogram (Fig. 11)~

Eight hours post admixtur~, the resultant spectrograms

appeared generally unaltered, except a decrease in absorbance

was again notea. in the MVI spectrogram. No shift in spectra

was apparent (Fig. 12).

The final scanning demonstrated significant u.v., spectra

alteration. The wavelength of absorbance remained essentially

unchanged, however, a hypochromic shift was displayed on the

amino acids/dextrose solution spectrogram versus a significant

decrease in absorbance on the U. V. spectra produced by the f>1VI

(Fig. 13).

Pi! 0

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6 CQ ,::q <4

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o:f Equal. Parts 8% Amino Acids So~ution and Dextrose (right) One Hour Fest ~ udmixtml Fig. 10. u.v. Spectrograms

in i'Jater 50% (left) a..11d l1VI

0) (j)

f.1q 0

~ r:Q ~ 0 l!2

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1.? ~ ~ I . I . . l j ' ' I . I ' ' .. -- I . : • : : -.._; ... " 1-1

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bl~• ;:~ f+~rtT~ ·~~ iWR-}~t 1l:rs~~~p,~.jL-Fig. 11. U. V. Spectrograms of' Equal· Parts 8% Amino Acids SoJJuti on and Dextrose in \'later 50% (left) and HVI (right) Four Hours Post - admixture.

0'1 --..:J

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1

:v-J•:c: ·,:; 277\ : ,,,. r· ·· · ·· . ., .~, •• 1--, r : l.tf: ::::.:;: w: ,;,, . A LrW' 'i:·:f'J' /i drc :: ..

H. . . : . I I : : . I .. : t' i. . : t;; I i 1: : j-1 ! .; ! I : max ·'. ;'' mr·· ' ' .. -:- : : ! 'r'.· : '1-:- l . : I ' ...:...::_;__--. : .L I : . : :: _l·

_J t .. : : . •: . ' ; : ' I i I : I : i : I I I I 265 : : ! ! : i : ' i I i : : l ~ . \ : ' . . ~~----:-r-~ . ----\ lllii'.. . ., 'I'' ··-~: . ..t.t'' f-iJ"". \ ... · ,., ..

1

-- 1:. f-l·. , :'L~-- .: .17:!_: -~~-: : 1 ~· ;:;tnt:~~: 1i1I-~n::,1 : .",~> .. ;::,I.:~L,:~~:-1 . . ~:: \ ~- -. ----~ ~~ - ~~-- . :_,__ ....: ___ : u_; . : : 1 :

1, 1 • ..::.;,s • : ' ·.:. i.Lf · : , \. : ' : : : : : , :: : : : 1 · .. ', , .,. · .. ·:· ,. .d, .· ~" ... ,· , ... ''· 'I' ·" ·n·---H· ..... ;1 .

I

!:::!:\,.::>·;: ~-ct!H+o-J !i Jf;.;. :~;i :[,! ;fTf:iH ;iJLi;~~· :~r·:-~. __ ::1_::: ::~:j!~~:~-j' i> ;.J:•' :''LJ!i\0:1\ii!lU 11l1 ''•+Hfi .,,,•_:: •' 'I ; 'F~;:::c1 : 1ilii~ .. Jd_:Jmn:mJJ~ ·\1ii ~:1JT::~ -r-- .J r~, · .

r: .. ·: -:::::!•·::1< :::li:: 1:::: ~\·.: il: r::1]· -... "'--~·~ .-~- --~ : . ~ . . ..•. !l' -i:t·:::--_(:-:·1::-;r· ~-~-<A:~:

-. ' ' :: . . . I '\. ::.:. :< i.

!,._t)-~[Ii;~;.:.i .ffi~,L=bt llitJ~H.i~if.tfi~Jt~ -8:-= Fig. 12. U. V. Spectrograms o.f Equal Parts 8% Amino Acids SolL.tticn and Dextrose. in Water 50?6 (le:ft) and MVI (right) Eight Hours Post - ad.mixthre ..

0'\ {):)

M 0 l2.i -='"' ~~ 0 C!J. ;::Q <!!

~91 1 ~ . _ .n Wavelength (nm) r-::::~?.9_ ~----~--::r::-~-?-0 .. -r ---.---· 1 __ J~_Q ____ .

'. ll T''r':;,:t~,~;, :Iii ! , Iff I .II ·!·: ~--;-.0-~.~~,1 ,rn r:\-~. lj"if!jt,c, ... TT:-T~M-.-i~- . I -. - : 1.~; • I • ' ·, I : i I I . : I ~~- --~-~-- I I . ' I t i I l' ' ;-:-- ' . . -I I ; ! • • : : I i : I I • ! ! I I I . ; I I l I I I ! ' I ; : I • ,I • : I I I I : : : : . ' ; . . " . r-~-·-:;7· ·~2----H--!.-:· .: .. l+.rr+~ ~--~ --1.:!--·.~-·-~--,-~-~ ~-;__~-~-- ---; .. --·--~~-------:._ ' : : . • • ,; 'r • : ; I i : I : I. ; • !' ; ', : ! I' I ; I I I. ! ; : . , . ; I' ' . I I 'I I ' I : : . . I ; I '. . • . . • I' I. • : I '. . I' I. . ' '' I ; . . . I ' . . ; : ' ; ' t : ~ • : ~ ' . . ' ' : ' I ' . I ; ' I ' ' . ' • I ~-r-- ---.----- --1.1-L-'--· -·--t--·- -~·. ·-·-·! 1·-1 ~ ~---1-~---- -- --~---- -~ -~-L __ '--;

I . . . 'I . I • ' I' I ' . . ; . I ' ' i ' . . ' ' ' I I ' . . ' . ,. I -l ' : ' l ! . . ! . : . : . . ; I \ : : ! i i ' . :. I I : : ' ' ' ' i I I I ! : ... , :. I . ! ' I . I .... : : I . I I 1 1 I . ' ' ! l I I • . l I . . ' ' . . I • I

' i . . . i 1' ! . ' . : ' : : ' . . I . " I I I I ! • I ! .I

hit--·-----·---_, 2-l--~ --~·---~. --~-:..:_1 ~-t.o .. L __ ----- -··--·-- '---'- --·-1-·-'-+------~-::.J . . . ' . . . I :. I :; ; ! :; '·I':' ; I \ . I . .. : t . . I I I I ·I . . : :.: . ! . . . : I : . : • : . ; ! ' ' ! . : I • : . It . I : ' • I : ! I . I.. I . !

L .......... -----.···' ............ -r--····1·+----~-··- ..... c:.:.; .. L. .. -. max j-· .. L !~_ ....... ·· t·- .... -·' ---1---·•·· ., ;._ • . ::. :·I·: .· .. :i' lj•·l*.l'i' :;: 26'7 5 ., I, .. ! : . 'i . I I I I · ·:· · ·' ·r· · , .. · : · · ,, .. · 1 ' '· • :: i: :l• :, ' :: : 1 .. I ' 1 ·. · · •

::::;: ·· -. ~--r~~.,::,: iTI j·:~ ·~;: ~'· 275 ~-9 ~H:;,~:;:: ~-A-.:_-·~- · : .·.. · j

·-n:j~ ::-~~-J-·:-~~:t::~:r~j~::~ :~:.:.: ·:r 1:11

11, r!;;~::r : 1 ~ H-1~·- ;j:: --~~;~;P.-~~--,-- ~---•+-~,-· ·•T'-+·+·H :-t+·r·l<.--· ~~- . ih r-:l· ,.1--! ,, . ...,.... --·-:· -----1 ··-· -·-·· ··-··•-'

H""ii ~H: :. iHH:!:·> ihlhl1,i: ::1.; ·: I 1'·1' ~. ·1 -.. ·: l-~-~-~ r·:·-·1 ~~:--11 ~-:·;-:·-: ''"1-:t: ~tut ""''·;;· ·: ~ :·· .. ; .,1, ~-i ·lr: ;·! '# .. ;: l-:! ;· ;u. :::.;II .... :.1:. '~-l-:. -!--~-: l . . : , !. : :; r : .. I \': • :: . :, : ! i i . • . . . ~~I ; i;; :! j: I;:: :::. . : .• ::. i:: .•. : •. :I --rr--- ~6--t-- ---'--- ·--- -.5· -+----..._~..... '-t- · - , !

,: :: ~·.::!: ;· j \: ::: i:!:: iili.:. . !i:Ji::f.:j,:;::: : :~ .::J:. I ! ···•1•· !''''' , .... I••\•· ''l'l"''' 'l'u·l·· ··-~/'··,•;1• ·'·.·!I• ,,j ... l. 1. .... 1 ... 1, c.• . .. 1 I' I:\ ,:; •!•:' 'I ,,· 'I'' ::; :•.:I ':: .. ::. ; :!.· ~-! . !.·- . ! ......... '"· •'I ' I'·• 1 1 ! ..... 1 :J-:• I .. , I . !•· . I t s 't ; ' ,:1 't

1 ' ' r--.~ -+i+! :+ +~~~-·-~ :·r• • ~ . 1 ... r I l I , .... !~,~, \ ·.· ,.,.'II', .. :, .. ,. ·i'·' .... J., .....

. : : . • · : . 1 ~ ' : . : 1 I : , , : • • : "max . ~ ; . 1 ; ~ : . j : .• i .- .: 1 : ; : : •• : 1 . ~ ; · . : . , : : . . : : : I f.-lt

1

..... , ...... , ..... ·+~-' .... 1

1 .......... -"ne. ....... J-·---····'··--·-··---'------------ _____ J . • .. ' .. • I I ' 265 . ' . I " I ' I ' • : 'I' I . t .. ; I ; ' +1' . . . I . ' . ' . I . . ' ' ' . . I • • • ' ' . ' ' •• ' ' . ' . I . . •' '\ :i_l . I .. I ..

. . l : ; : : : : : ; : : : ·7~ : : :: .i < i ~ . ; : i J I . : : : ; t· 1

; l : i j : :. : ; .;f': : \; ; : !7·-· :. : q ~;:: . ; : : ------.\.-~-.. --·----+-·-·-··u'- ,. 1 ···~·· "· HI . n L ... .,yjDl, .. . .... I:; + 1 ; j':•i'!:,:::H~!i'' ~:-\iii::!' ;'841\ I . ! ' :)

T;:x·r·t:-1 )I '' 1;:!' rtn: · :· ;·f~,!:rr:·M- · ,~1 ·: · --b· yj +·~-+II H r I ': t I': l : ~ : \-1) . ! : \ : Til i i '' : I' I > r·-r i < I . : .. I ' . i .... ' ; .J..J...i..:.. : . '~ ' I . I ! ' . LJ . I ... I . . ~ j i ;

t!: .. H,J+ bi·h tffit~,:, . I~]; :r:~ · ,, I :,'H,rmr.ar Fige 13. u.v. Spectrograms of Equal Parts 8% ~~ino Acids So~ution and Dextrose in \'later 50% (left) ano. NVI (right) 24 Hours Post - ad.rnixturd.

m \,.!)

70

The Beer's law plots for Jlol:vite, Rubramin PC, and·

Aquamephyton confirmed therapeutic admixtures were not of

sufficient concentration for scanning. However, in order·to

determine the stability of- amino acids/dextrose s·olution,

each of the supplemental additives were admixed singly, but·

only the amino acid content was scanned. Corresponding

intervals of examination employed in preceding studies were

utilized. Resultant spectrograms for each admixture failed

to demonstrate alteration, suggesting an absence of chemical

interaction.

?1

Antibiotics. The determination of standard u.v. spectrograms for the selected antibiotics indicated thera

peutic regimens contained· in the amino acids/dextrose solution

would require scanning of undiluted solution·with the exception

of Keflin.

After repeated determinations, the spectrogram yielded

by Polycillin N, 0.8 mg/ml, was difficult to evaluate. There

appeared to have been a \nax of 252.5 nm. It is conceivable

this deflection could have been influenced by analytical noise

(Fig. 14). Scanning of Keflin produced a A of 236 nm with max shoulders at 262.5 and 283 nm at a dilution of 0.02 mg/ml

(Fig. 15). Production of a spectrogram for Garamycin required

a.concentration of 0.160 mg/ml yi~lding a ~ax of 256 nm and

shoulo.ers at 240 and 280 nm (Fig. 16). Similarly, a Am ax of

256 nm appeared on the spectrogram for Kantrex, 4 mg/ml, \'lith

a shoulder at 282.5 nm (Fig. 17).

ri! 0 ~ ...... l=Q

§§ ~ <!:!

250 .,., 26L 220 ..

Et' I tz.o~ . l· i·i·:· Fij:; lnTf:;:i -, 1.5-t-·-i.· ·t ~~ ~~;: ·.:~:L:ii!

. ---L-12-;--- ----r ------~·-c-;-- .. 1: 1 I.· i i i: I : ::: I: l:j·ll ::J:!Til I---~-- -r-U--~--•--!· · .. +- -~----F·-- i ~ -+:~ .. -t l I ; I i : i . ! : I l I ! <: I I ; I I I ' ! . I " . ' ' . i ' I ! .

L---• -: ~--1-' () .. ; -·---:--... +·-~ . ---· + ._ I .. ~J ',-'-'.:I L\_.J ...... , .... !----i-----.i .. : ... ~-J:_: I :;~1:,: I fL t· i-t~--Lj,. -,·[~ r-·.-·t·\· c~ ·o ·-:-,:t-:-.,- -~: ·:·-:·;-:: -:-.--:r~f'

F ! ~ ,"'i ' i .. t> ~ 11·':. --'"L----f-.. ,. _, ....... I ... -j . . ...... 1 -+·· .... --+·--T---irt·.: I-~ :.:1: i .. I.· 1:; ·>t>. ·-i. . . . . 7 . I '\~-.-+-:-tt- ~7_;-;t

1-. ~.:; ---- : .. ~ .. ~ .:\! ... : .. (.~· ~~ .. ,.~.t:..L.... ... ..:...:.:l.:..L ... r+ 1 -+-11 ~\--f~-- 1 -.~!'.: ·y_:_,+_:L

t ... !. .•. , :. ; .. . .. i .. --- 1-·--1 .: --i : .....

-• ·_:_. -1--$~-L~~---L\.-~- \; \ ·-r---J~-h- f- -1- i i -• +.X- -l: r- 1 •. J '· t-·~· -~4-+---1' ...... _. ---~~-~~-· ·-+-~ I.:. : . i : ! . ::I : ! : . i\ I:; :j :; ! I:;;:

.... -. ~- --~-- .. -. : ... I ... . 1- - ... - \ ~- ..... . ·-' ....... ,

~~-~-- -~~+~..:-!~-~-+--~- _j___~~~:' _:' :~~-·~ !'~ ~.;:·:_[ ____ ~j, __ :l· ,: :;:J~:_:·r'•~ ·:uL.:.:: - -:. : l . : -. . ; j:. : .. --.:...,:. 1: j::

,·· 1--!···i' .i··· ···j~ ~:,

t-T---~-~t~~-T::t~ 17!::: -~~-;-1 -}~;_ i- ·:-.! : ... ·:: .· i. T:: ·:!·:' t·-~~:~- ·--}-··:---1--:~.T ---1- --~ ~:: l- :_f:~: ~--:·' . ·t--- : .... j .. :.; , ...... ! ....... 1-. ' .,. 1.::~-1: ~.J l' t _.:• t::, LJ.lL . ..:J

Fig. 14. u.v. Spectrogram of Po1ycil1in N (0.8 mg/mJ.)

\va.velength

A max 252 .. 5

,......_

s Lf\

"' (\J !I\ (\) '-"

f:r.1 0

~ p:j

gj 83 ~

<:::ltff~:~~11RillfP;~~Titfh~ • L.,.r- ···r,·l· .. ···- ... ~ ..... "U -. ,_ ,_ .... _..! ..... , . l. ! .. .[. .. , ... ,_ ....... ' - ,.. ... !. "'-: .. r

-~T:,: .. r:·+:, .. l·::.i-TTl~ : __ l ! 1.: 1- ···[~i .. !·r:~!·r-l 1:i; ~--1=1T ... t i:I~~~Twi •.. ~I-Ll±'-···urH·

. sq_ -T- t--·· --r· H··· , -· •. ·t··-. ·~-·---j~· --·~r· ~-- I r ' Ill ·I··· I 'f ' '}'Jj::j:'c, __ L. -~~

~~-1- 1:~1114-t~litBI .2 .4 ·f .8 1.0 1.2 . 1.4

rJJ:i11igra:ms/Mi11i1i t er Glraph 7

Standard CuJve for Polyci11in N

"

--J 1\)

r£J 0 ~ ''< (:Q p:: 0 C!.2 p:::J <4

200 _ _g50 300 \vavelength (n.1n) · ~--: ,._ .. , .. ,.. ~----r- -~-- !"Tr ~-- n --r · ,_ ;--~- -r 1 -- .... ,. · :-r::T ~ ·: ·-TTl

-- ; :q=rm+,'-I T':_FTI"' 1-,--jl ,...,...,...,-~ I i! i! II' 'I

y: f1!i lj!i ~~!/--;-l-++-+~~-'-.;1 1 . : : ,. i :: : ~~ ~ :, ~ :-r: 1, i , : i. ~ j f-,

. I· I ''I I i. I II'

I > ~·-·--- L i'lj

· . . . . _ 1 · 1 · , -------r- --1 1-

j :: . ·. ' : I . i .• l! l; : i:: :: I; : I iiI J",d'l t-~t::-1 :' .' :HT H '_ ·~'-f-: -l:4 I I- .. I ... ·I· : .j· ---I-,_. L ... . .. ' :I I 'I ·I· r; ... "

.· 'i I T • : ' .. ' !!: i/!i 'i[;i! mr I'" .. •'... . > .l+M-. 'riG .. I. ., I,~·.. '·-·· ..... _,' ' it' ". i

1:,··-'·-- tc·· ,----- ··'--~~_:_:.: ,,,,,,,,. jl, 'I'' liji 'I"· I . '" · • .. 4 . ··-·-·· -•· ' ' l' 'i+' I tf

liT< f:l J '-!:::!if 'i!pi:r lilt>-1 :il..+-~-'

1 ~:-i~r·~ ~:...:_ :_ ~L~.J :_L_;_ \Lt! !I!! 1!!! i! i i i i 1 1 max r-. ,:!:::::;·~:l: .•:: ::: ~~~: rit: nt~-n-+ _+-,-.:.L-1-lf 236 p ~ j/ J'l ·: , ·: i:'! ':;L~! ilLJl\l 262.5 ~

1-~~JH-:~-:-::-'-.:l :...:.:.J.:.L ,,!: :;: .::: .:.;: ·:'! 283 i<\ ~::: /LW i i; Iii,- :j-;-, Tl~-- ~c-j-•- -~~.:_- .:..:..;J. (\J

:jj:i) I i •.' < I ,· :::: ::; ::: ;

1

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CIDZL:f-~T-T~~H+t:ii··~IW .lil ; I : :; : ! : : : - . : ' 1 : . : . ; ! ; .p.. : I i ! I : I : i ; . I ' I I • . .4+ -,

lilll[_t :Ir :1[rl_~ m~~:: i ,lfj !. ! :! I i ij~ l . I . ' . ' I ' - J,,, I . I < . . ...

J!ii~• ~r: r:u•;: • i •-~ilti!J:ji*~* :,~ I:,,.

1::.: ·, · •... 1: ni--~- -++-"-'-.: LL : 1 'Ll .. 1

, :, 1 I

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'--' . . ''; ; : : ; , .. ' I I J i I 1- ' I I' I I r . '' · ! • · • 1 I 1 I J ol -r.~• 15 ~r IT S -,.,.+- "" .f K .fl . .,., jj~g. • U§v. pt;"'vrog_,_am 0 -8 ~ .. .1

(20 )fg/ml)

..... "!"["; · :-- ., .. ,. · --~-1 i ' ·l '· -·:- · . .! ·· ! ' l '· 1 · -·:- -·

I_;:J --. I I I ! I ! • t \ ! I : . I ; . 1 : 2. o~-~--r:--;- --!-, -+--lml. : +-: ~-· 1-+ ~ ....: - '--·-"--- r--"-- -----f--;-.'-~ ; j ., ;-1·' 1---:·j"·!~ ----~ell-.(-!--; !'-~·! ·; ; . i ;-!·-: ! ""(:.-~

-"- L t .. 1 ... 1-r: -:--, -rr-' -:---!--,- _, __ _, , ---- L.-'-- -r---- __ . . : .' ! -I . f ·I . I . - !. I . II : .. l ' . -- L . . ' -; t : . ' . __ _j

1 8 i. l. iII l I i ... I '. : • • • : .. ,--,- I -; I : , __ • : .. ' - :· I ... I '. !· . ·- ; .. : ' . --- --1 ' : ' I . I I I ' • . l . : ' . I . :

[··,· .. j·l . ,. , ... : ., '! . . .! ' .. ·-'

_(_L, _____ --,---'1---i-.L ... .: -'·-·-" "_,_ ---1--- ~._ .. ____ L__;J I · i · · · • 1 ' ' •

1: ,-, ! : ' ' i ; . ! l I ' I . . : . i ''l . . I

-- . I I '· I . I ... , - I I i . . . ' . j • - ·- • • ... .

1~61·-L-f.l! 1,. !- .. ! __ ~-~---~'-----;---~·l··:-i- ... ! _.' i--i-- ·_: ·- , ..... J I I j I ! I . ' . ' .. . .f

f -!. ---, .j ... , --~-·-- -. ·--'-1"·~---J .... c.l--- r--· 'l -- : . ..,.-------' ~- ---

f I. . ! I r. I j i ' I ' LJ' I . I . ; • ! .. ·' J I 1 I 1 : I ' ! I r · · ' : , • · · ·

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1 • 2,t-1-fT ~-1-- r --!-~;lfi "TTT n--~---:rT~--~·-:;~~:~~ l r--r-.:4-- '--1--r--· ___ u ___ ---,---r+---,--- ----~-- + ----· -. ! I. I I ,. 1 I ! i f i [ : I ; ! : J : I : ! I • ~

1 Ol' : I . I i ' I .... . I . I --. I • I . : I : •• ! ·-;. ~ ., ' i ---1 -. ..: I ... ,. r--· .: ,. . !. .. I .. ! :! . l .. ! j-! - f . . 1 l __ ; --l +- I -'-- . -~1--..::J I I I I I I ' • I I I ' I • ~I I ' l ; -~

~-~~~~L-1: =t -J ·-~!n;·=r~-- j~~~ -~~ :J~ t. __ ;~~-} --~-~- -~ ~-~ -~-=~--l : I I I' I . "l' I ' . I . ' I . I ' . I . I I 'I I . . I ;. ' ., I ' ' •• ' .. ' ' I I .... 1 I I ~ I I I ' 1 • ' ' • • I 0 • 81. .J .... , .. ,_J .I_ ___ ,- ---1 --'-f--1--! 1- ... !- '-· J --- • -i- .,.. .. ! .... ___ ; - ----l--

!-:··· ·1 :·+f-- -!-L:·-·'-·-;·Jr-(r--!-T: _, · 1 -~---~--:---;-r ~ --:-:----t 1 I I I I·! I • ; i ! : : . ! : ! I !

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• -!---- -·--+ , 1-- · 'r-' ·- f .,..,._J -,_ -~---~- i---- t------·r .J--- -W-----l---. : 1'1 I I ' I I . I : I I I i I • : j ' ' ' ' :· I I· ': I l I t I : • I !··· . j' I t i ... f· ·-·

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1-!- - ·- f 1-- : ::·~!-- I_~ -~ --p-L; --1..,-J-l· -:- ·;- .---. -+-- 1"--f--r -;--r-f::i 0 • --j--- "T - r·i--i--+j __ :-- 1 --,..i----t--- r--~- i- "- -1--:- ,---t--;--H·--' -~-::"·-!

,I I I + i I" i ... 1 ! II' : ! I l ; : : I . : l ; __ , -; ! .. l - . :.{ I !:·' l '~·I -~i I :1 J I I ·I I : I I ; I I _j .· .. 1

L i-T--: .. .-:-: -::r:, r,_r ·-:r:· •i ~-:-~· -t--t--, · ., : i : · ~-' :;.i _I~ I :~i;c+---k,-'-!B~-~- ~- +·-lJ + lLi--'i--::---- 1-!- -~---:-i--7.-i--i· -,-1 --!-~-i'j I _l!U~:.!.LL.L±: . . _L -'- _ _:_::L_l_ L-' _, : ' i . ~ l I : ;

1o I 20 . 30

Microgr~:i:;~i~li1i t er

Standard cuJrve .for Ke.f1in -..J \,.>J

·'!

~ 0

~ p:J

6 w p::J c::t:

220 = 2 0 . ~ 20

'TI1Ti. I!; '. t • , t, I I + Wavelength -~ ... ~~-~- :·~, •• I!! 0' -J·I·· :''!I' I: I I:! i I! I! i : 1 1

1 '1 I'

+-'-.. -;-: 7-~ :...,.,-,-~:1.Sn:i : rrti: ~~ : ~ rTn-r --- -u-" -~- · +----...,..,-"-L

l. ': ' ! ' i: ' : : '!• : : : i: :! ., :: 1: l: ! l ; I! I! i 1'

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~----'--~: ..L; ~_]_ -~- ! .... - .: -----l~-· t.:.:_ ..

f~~ · r~~ [: _, \ -~1~. !" -! -r)Jtl _; __ .i;.:,.:.:.:l - ~ . .:. .. l: ... j .. :.c.., ';[~:...:.

. ..... ·t l :•""1 ' 1'1!1

: , ; 1 'j :r: FiJ.~ I '·· .. ~·,,:·it:· ··i·l·• l .•• : i: ~ --r I

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~---jJ __ , __ .. -t-....-· .. --r---~-----1 :: ' : ; 1

1 : ·: : \• : : ' I' I I : : : : :1: \ ; I·. . . .. l- . I . I '':' '. , ... ,., :''

L.·_· ~---r -~~-:-:-~.:.\:--:~-:J. __ ~J.;):::. I . ' . I l . : : . .. ' : I . I " I . I

I. : : . ! . :; I :: i -: : ~ . ~ I -: ; L : . < i. 1 I : ·: i :: 1

1·\ : • l . · . :::I:::

r-..;..----+-r 1-~4-- •··.·--· ........... ~ .... ·----~~~---' : : i : : j' I .. .:; : I\: ! i :': ' ' : : : ; : '

t--·--~-- ... · ·c--f --•-·1·' .. j ... ,. · ..... 1. • ~- '. I·: .. i' ! .. , ' i'' . I : ·.l'' ' '. i: i: I ' H+-1 >pi _:_J~h~~jt~tP, i: :tt:.._

1" ___ ... - - ..... t. ..... .,\ ·--. -- ......... - ... . ; . ':~ !~:- ... ~ +l--~+<~---~~~-111 tl :[ j r.-;·-~--·~~ ~; ~T~~~-

1~-T ·T-~} -;·:·l~~~-

1 ..... ! .... ... .. ... : ... ' . :: I .. , ... 1 " ; I ' ' ... _: .. :::.:.L~l:::_.!:'l .. ~ _::::l:~:·i

A max 256 shoulders 240 280

Fig. 16G u.v. Spectrogram of Garamycin (200 _.t(g/ml)

;--..

~ \.0 t.r\ (\J ....._,

Pi! 0

~ ~ ~ 0

83 c::t:

.:: '~L-.L.J ~-+--~~.;__L_~--~-·-f ... f-~ -t~l----~--~-++·t+r-: -r------t-r n I I [; ·I , 1-.. • ' , : L ' ' y.J., .. : . ~ ., .. 1 .,. •• , -; • ,. ,-+ PFf:i :,-f-fl--f~l~~ l .. l.,rt+rt-i-rht1+: .i :--~~ttt

80 s 160 240

Microgram fi~illiliter ~~9

Standard Curje for Garamycin

i~

--:1 ~

M 0

~ §§ ~ -<

f~+ I 2?£= I • -! 5 -L-~ .-L-~.J · I : 1-t ·-, 1-. ;~~~.ln~P \vave1engt'l.:. ( ) · I .-·--~-' · 1''''1 1 1' J..t nm ·

, -u-; _.,.i l ! i:, ; I :-.~j :. ----+t:~~ . ·I ·--:--........ . 1 , .-r--- J ~~~ ._ 1 1 I J 1 1 ; ! 11

2 0 .... ·f · ·' - l·j .. , .. · --·1-· · ........ .. : I , I ' .... c.~ ' • .. I ., ' I I --I· ..... . . . . . • ' ' '' ., - -- - ~ ' ·, . ' ' . +•· ' : i · 1 ,---~--·---'''

1

.... .i. lT:-·Tj·_J. .. · ,-: .. ·· '·r--.::' I ''i' · •·' , • ,. . . ' . - ~~----;--7 1 8 ·r· J! -- -,--~--;--.. -~·-- .. _1 I :-I : ... L.t ·i _;_~,-:--,--i--r-;-!---- ~~--i .,t· ' . • . , ' . • . , ., . ·I.. ' ~ r .. ,....... . -~--~- =-i . ·I -~ , , ·:· - •

, .. ,,,_. ___ !.. ' ' ' I ''' ' . --~-i- .. ' .. ' .. , . II ' , I · ... , ......... l.-! ... 1.' ·- i""" t---!---1..

l- -~- .. l .---:. -·,--:- 1 ; .. · ... 1 .~·~ 1 rJ i T • '·, ..• ! : i _:.: !-:-ttt : '' ' '.J I'." i ... F I' .. , ......... L.i ., ., ,_, .• I .. ,-· •I.e I.,·· . 1 ~-- 1 1,.! 1.6 .· 1-[: .. I,,L,-,I.._, __ J __ ,i

1'1,-·

1-.. ,., ,.,.,+

,_..., ' . ' . ' . I ' 1 , I , , - - I ' ' I . . .. ' t ._,.--- -~-'-~: ·· .J·: '' , -· 1 1 .. r-t 1-+----!'-I-:!_J.i 1 · • ... , ... ~r! :·: .. +-L+l'.+ ~. . _,. , .--- __ ; ,!_,. " L ' .. L .. ,. ' '~-·-j-· 1 ... • '.''' '' ,.

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- IT , . I ' . ' ,· , 'I ' . -' ... ' ' ' .. ,. . ... ' ' . -- ..... ' ' .. . '' ' ' I . - . , I ... '

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. . I \-r- ,1,. 282 5 . 'I _! i --~-- ... , .. , .. L,, i' ~-··'·----~ , r·-;--"

'.-:··I : i ., I,.. .'i'l~-···- • --~--- ,--1~----L ... i· i i .• I:~-- ·r ; .. ! .. L:. I L ~ . . ' ' ... . ' ' ' • • .. - - ' - --·- __,_' • ; i . ' . . • ., .. -r~ 7 t---rL~_l·' \ ·r!·· :! : .,1 ;::''"! s L 2 +.t--r1-. : ... J-1 ,~t 1 ; r·-·;t:'-1·:-+-~-+ _ .. , - ~· ;-t·.

! . .·;- --- ----r' ~ . , : ; ' ~ ' ' ' , ..,- 1-: .. : .. , . I ' - I . ' : ,-.~ -, T. : I . , --- r ·------'' ~ . L _I ' i ' ., ' . . t I .... ·- . . . . . t : • ~-. I , ' .. I , ____ . , . " I ' I ' ... ' ' . r j . ' , . , ·,. ; · , .... , ... o 1 • , · I . . , ~: : . 1 ! .•

' ' , . I <JO --- ' . . .. •· , I . ' ' .... - ! ,, I : \ I . -- .. i : ! : I l!\ 1. o .. , . . ,-, ,--- -:. I . : !. _,- --: - J . . r .. - .. : _ _I' i , L :_ t ~ t

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1

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: .. I· j ·; ! . I!-; ... :~--~ ;·rl I I . . ' I I 1 , , _.___ l ....... -~ , . . I J. , • • .. I I ' I ... , ll : , : ' : ' I f.i:l 0 8 . : . I i" --·: .... ,. . I- L: , ~ I . I . I : : . -l . ~ -~ :...... L

f\ 0 .:. - I : ' . I . '[' : 0 • . .', : ; ' . ' ; i ' ' : i-·1 ~ ' : • : q ' ' , .... ! .. -- i\... I I '' ' ~ ... , ...... I ' ' . . I ' ; I ' ' • I • L .. : ' ... , ... f- L ,. I \ I I i ~ I'\ , ...... ,_ ... '-'-' I ~ + .....: ... '-, i . : ! ; I . : . ! •. ! L.: .... , : ' . I :. [ ' ;

1

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. I ~--1-,·r-~-~-'! T---f---~-. i •. ,;j:jj I. ~-:·-rL-"-,::L ....... .:... ... ___ ~·~~: . :·1 ! I_ .. -; .. +·:_j~,H J·~· .:I:U+i+·;=cr:j::ffi

• . I " -· -- • , I I r· . ' ' I , • , I . . .. -- . ·"· ,,

t---.. -r-~- --1 .. :....-l~ .. : ...... _.__ \ :; , I I.~ -:-~~ o 2'. -~~. 1·-- -il I I' , .. t ' i ---:- I.--!--~.:.\- :rt~L+ ,~.. L- -tl.

z-' ---~_: !.''. l_;' -;--·--:.~ .. " .... , .. J-----1----=·r·j--+t .. i .: ... ',' ;---:-·. l .. i.·;- "f::''" : I --. -- ' ' ' • ' ' .. ~- ' ' I -.. '- I : : ' -·- ·-t-· , .. '' , .. -,~ -"- ' ... ' I 'I ........ _J_L' ,- .. ------!..' L

-· ; I , , , ; ' I , ! , ' . ·--~--·· '. ".. - . ' . : ' ' -;--+- . I ,... '

' -~--:_ ---·-·! ·

1 1

' 1 2 - I__

1

"j .. :.:< o·t-~ ...:.:LJ !-~1---r-· ;....l i · · 1. _ ... -- l'' 13 ' ' ' .· -;~Li J. T

--.-1 . t·:· l . -·'- '--- ·:-i·.-'o" - . ' ' .- ... ,, ,,

0

I ... ! ,· : ........ , .. i' - ' • I "I , ..

• t • • • I I , : : : . \ • : t ~ l -

Fig. 17. U.V. Spectrogram of Kantrex (4 mg/m1)

Hil1igraJms/Mil1i1iter d

1

raph 10 Standard durve.for Kantrex

......:J \J1

--~~-~-----~--------~~-

76

Having previously established a Beer's la\'.r curve for

each antibiotic, single admixtures of each antibiotic to the

amino acids/dextrose solution were prepared. Prior investi

gation indicated, with the exception of Keflin, that thera

peutic concentrations were of insufficient strength to produce

u.v. spectrograms. Consequently, only the amino acid content

was scanned 1, 4, 8, and 12 hours after admixture.

The U .. V. spectra of' the solution of' amino acids and

dextrose containing Polycillin N, 1 Gm/L, appeared unchanged,

except f'or an absorption peak at 224 nm. This secondary peak

remained consistent throughout the 12 hour period of' examina

tion (Fig. 18) ..

Upon examining Keflin, 2 Gms/L, mixed \IJith the solu·tion

of' amino acids and dextrose, a hypochromic change in absorbance

t,ras noted (Figs. 19 ana. 20). The· significance of' such an

alteration in absorbance remained unclear.

The U. V. spectroscopic examination of' Garamycin, 80 mg/IJ,

in the solution of' amino acids and dextrose suggested an absenee

of interaction (Fig. 21).

The final examination of' Kantrex, 500 mg/L, admixed to

the amino acids/dexGrose solution, appeared unreactive (Fig. 22).

To further evaluate the results of' the U.V. spectro

scopic investigation of' the above admixtures, a microbiological

assay (pg. 86) \vas performed to provide a more definitive anmver

as to the possibility of' detrimental interaction.

i •

j:i:J

g ~ p::) ~ 0 U2 f:Q <:

2f0 . i __ , ~: , " Wavelength (run) 2~4 , : ; ,l.l_'B__!...c.. , '2g , 1.,,,; , . . ; ~~w , 1 11 ~ 1 m _ ~hln 11 !JJJ! ~ , 1 ~ ~ 1: ~: 1::: ~ . . · u ~{ LL-li·i! 1!!!:!! I JliT!Wll! Pi~il!d!!J!i 1!:. ii!:j Jii !. l~!j t. I;'; I . ';I' . ' "i' j j': 'tl'li' I I-JIII;.; 'II'. :'. . '' 'I' . . . I.Z""'T

tl : : : , , : ~ 1 1 : 1 ; : 1 : : , 11 i , ! ! i : , 11

1 : I 11 1 ! il 1 1 1 1 1 : 1 ~ i 11 : : : :r : · . ! i

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·I·· •.. t'. _, ... ; ' . ,.t. I . '.' ! '.I,, . i' I,, III i I I' I' I'';' 'I:,,' I ; '' I' '·I . . \ I I I. I, 1' I I Ill I' lo ], r r! I J I·,· I. j I :·: ... i·: l . ; • • • .. :

·t.:>il.<i::' i,i <:!;;;! 11-t:'iii: if+;;;: :;h:::;''·i;;-::: :;::!:~·: -_;.~ - ---~~----·-I·-· -l -*·--·1··-,:-rJ .. ,. ·TI······- .. ; i·····~ A . ; I ]I It·': : ,·,!· i' :·•. '':'. " I·':. J. .. .•

jl :! i : ,, ·::;!:·.:l.ijW ;I: ~~3 Ti',ti!T .ff'U~ i~if}'t-;0!T~: ~4 .. , ' ·I ... , . "·11' .. l.,,. ', • I· I', 1:! r9 ... ' I,.,. , , . ', , . , .. ' . . . , . I, ... +Si ... - ---- .... ·•··-·--·- .-..----- ~ ·-·~ ----·: _._ .. ~- .... •· .;1 · ~ ! .i. i.¥, ! i ' ' ' t' f-f''' '' j: 1_ ' _·

1 • • . • • • I •

· :: ·: .·.;:.:: :;·. :;,: 1;; :li·! Shoulder - r,·ir'l:rr:r:~-~:·,.·i:·: ·~:'"!:t<t'l'1T;-~·: ···· ... ~ : .. : . ~- : : ! ·B. : ~ : : ' ; : ; : "i-~~l: : ; ! . 267.5 r+W-1 : i i I 1-~i I : : . I ' . : : : I . : •. I ; : : . ~~~

jt ___ ·I ··J···· ... !! ....... ··~· 28'7 .1!1 ,,:. ,.('11 .. ·1, ... , ....... _,, .... "

~t~;-:· fi-:-::t·::..rr~~: :+f1i;; v:··-- :~it·· · '" 5 -:+·: 'i-H: H-tH!:-:~'rJ~~;;.;;.: -~L!-~--~ __ _:_:I ·t·· , .. :.! .... ~,,,, ·~•·li!l ,Jri i\·~ • l''r[ljl' '.\'!'''''''I' 1:: 'I'!'•. ··J r·. r:: :i.i'i:: ;;: ~::·1tn-r1ii -~~i:i: i\11 :;;: ·~:;::;:··:;:' l' ·: !i1 ~ ~~·t-·1·1·-· .. L. ,, I, . A "' l·j .. It ' j .. .. t ~-:-.>.:. :.·:::':':~: !:;_ ·~~:l'l!~ max Jjj:·:r ,-:r:jl·Ti:i:·T::: ;;·;~ : .... -·s ·•., I • ·' .,.,., .• ···r· · .. , .. 'l'mml 224 '[''''''II. ''I:,,, .·.;.,,. , , ·t : I . :~ . . • : • : ! . : y. . . ; ; . ; I . i ( i ! ' j • I : 1 l I ' \ ' . ~ ' ; : ; ~- I . i i : l . • •• : :

·!--_,, ___ , ..... , .. , .. , .... ,.L,,, l. l!!·,··t "'79 . ' .. hjl·'. ·• !:_ .. ,!·,,\.,:_· '! .. tt· .. " ·'It··.,-~····' ·I,'·'· "' . Uill' .. i .,-. , .... 1.,,.,,.~ ... , ... ,\ .. -·- -+----~r-·

t-'4, : : · : ; , ;'. : ; : i·H ; : ! ou er , ! : .. ~J ,j'-L . : :: :\ ; · · . ! : .s: ~I· . > ... ·: : :. 'iii\;;; j,;!,iili Sh ld i •\ iT ''j·':l ·1:;::' ! .. ·,. i' .. :

I '' . J. : : : ' . • \ ; .. i' I I' I 267 5 I I I :I ·I· 11 ' I " '~ . ' . : ' . ::I 'ffSI·~~-:._' ~-: ·-~~-H +;\\··; :·J: .. ,L~.tt: .. 287,. 5 .~ :.:.1:·,- :_: ;_ .. J..I_/1'. ,-. ~· : ...... ! \.·I·-~-, L: _:_ ~-·-:_j . '.,, .. ' '' . II. ' 'I I:' . • ''I I . I'. . . ' .. '\ ' . I ,. . ' . : . ll : : : ' ; : ! I : : ~ .J.H i -~ . : ! ! : i ; i ! l : I t I l : : - ' : : : i : \ :: ! ; . : . : ' ; ; I I ; i i ' ; j i ! ; l '; r i i I : : l ! I i 1 I I . -:;rtiH7. ' . I I • • I ' . : . . -~ .

. . ' \... ; T; I i .,, ; ; 'i i I' \: ; I!: i i;' I l' :. --------l .. ~ .. ·e-- ....... i·• •·~11 ·-· . ;;!J I ,II• I . : 'I. • ''1. : . . ' I' I. I I I . ' I I I '· •I ' • • . - - .

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1 ' ~ : ' i ! 0

• i ' \ · I f ! • •

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,,,, .. l·,i_JJ'' ''I' 1.!•1,11~, ltttl, 't'i· 'il' ,. ! '1'1"''· "' · . ~ ,, . ., ,, ,• .. '· ,•,'1 I . 1 1 •:1 1 I:, "II •1 1

• ~ • ·"

~~ '· 0i:l': ~~~: ::;-~ ~-qn:rn: it!:; !i; :;~~-: ;-- .'!;::;;. ::: ~-~·~: -~~-=r . N4'... . . I' ' •'>.. ' ' 'I I I I .. 'I'. " . '.'. . .....

--,·--·-~----1·-··t--!·· .. ( ........ n . ..~ .. , .. ~~~ Jl1~~~d·j·iJ· ~··· .1.-~., .. ·~~~ ..... ···.il : .. :. l::.. . : .~;:: ~- . : i: ::.;I,;.:. i: ~ ~~ ! ! ~-~-f-!-! ·r _·.w:- ;:-· .' ·rr'l :T- ~ .... --~D

-- .:....;-i-~...;.:._f..:_' ..._.:..:.. ;p:.t_u_ ~ • . · i I . l ~, ,_,' 1: • • ; ; • • l!: ": 1!,: ·: : ·.: u± ... ,!,,,,!.,!. "·' ,, 'i ,,,, jll'jl''' ·I· I"' ,,,,,.,. _,,.1., . •j · ;_.::.~.:j::;·,;:~ .·~::;:::1 . :r~·l-i:T: ':lu -:--·: .... TFT:~·, -~T:·:_· .---~

1 hour j 12 hours ()

Fig. 18.; in Water

U. V. Spectrograms of Equal. Parts 8% Amino Acids S.ol'!ltion and Dextrose 503,6 in combination \'lith Polycillin N, 1 Gm/L.

---:1 -..J

r::£l

~ P=i 0 U2 f:Q ~

-~~--~~-~~-~--,~~L2~9~-~ • I : 1 ; ! ' l ' ~ • • I .j !

, ~ •:.1,.,, 1 trt,Jtii r~·j, 'Til --'-..;....;.+---4-4. -~~7! :+++-: H.~: ; ·;;

-'-+--·t-{ '!:hi! t::idl\! \!\~:!1! j·: 1 ~ -;--T:.i ~~.~-~;r.T; ~-~·~;-ITT! I. I. ! '1''' !:·:·I! ;·,ij· I'

1--·1-----.,----. • : . + _ _[_, ___ ~ ._; :_ : :..:.+~- ·_:.;_~ .:..:JJ_ \<l •. l' !'·!;::· ,:!'!'!,: ::: ::: i

~--:-~ ·-------1----t-·--- .......:..---r--... J.-1 o~ ---• ... j \: .. !::: . i. , .... ::~ : .. !i!::!;~:~~:~!:

- \--t--: I~----~-.. -- ·r·· :- - ·- :··r;:: · :~ ·1 ,...~-·

:idl i i .~ ... + .. ' ·:-:--r-r.~- _____ [ ... ~ ... : i--~1.....:.:__ .__:.i. ::1:!:~~ d~_:'.! ::;. ill ~! ·:: PL:. -~:~ ::::;,>: -~~-~~·:-l~,i'~· 1, .. -l' ' ... . . . . , I- - I . ' ... I . I . .. . . . .. ' ' ' . . : . . I . ' . . . I : I I ' . ; . : ~ • • . : i . ~ . . I . I . . ' . : . : ~ ~ : ; ! : ! . : l ;

·c· . !\ i/:--r.:.: l. ~\ :; \ :L:. -:'~: ,-:r-n -- -·· --- ---. l ---·- ---- ... , .. --~..;.~ . ... --~ ' ....... ..:. . •I . . I .. ' .... , .... ' ' 'I: 1 .. ·I\ . 'i ' . I' \ ... : . .. . ' I' : ~- ,f; :: ·: .. : . :;!· :·l~:

· · · i · \ · /:I · · · .. 'j \: :':: 1•.;: ! I l

F~·:·!-:;·~ /~~t~~~ ~-u: t;'t ~;tTf:. -Ht ·i< !::: \l}::: . I·

bf-- I :t'r'tt-. ... , Fig. 19. U.V. Spectrograms or in ltlater 509b (left) and Keflin

Wavelength (nm) 20Q .. IT.2.0 I lll -,! t

250 300

A max 268 279

~ax 237-5 262 .. 5 277 278 282.5

::J:I!ltlt':_l''l:::li·''·';I.'''H·':I· ~~·~ I I! . ::; ; :-1: ! ~ :I : t:': .· . . . . ' .

1 \ ! I ! . : i I I I ; 1 l j ~ : : t ' I ! ' ! I t : : : : I ! ! I . ;

r~ 1 r i 1 !j: 1 i i ~--r :··::·-~- -. -, -~- ·---r·--. · : ! j i l I I : I j l ; ! ; I : . : ' ! : ~ ! . I !

~:·~: , : :~ -;~1 .. ,.-i. r ·;~~-i--:-~ -~G--· ----r·:--· 1

::.E ~J;,'· :~r· ~- L~- __ L ________ L_ -sf+ ill: 1 i -, H , 1- · : -

• "I , 11 · r tEE·

DV<.rt: ·w,N _, I I P1 ~ .:.L .: ' J(i 1: : ) : f, : ; : i . . !: : : ; . : : : •. ,. ·~~-- ..L:~-·· --~- -r------··· ·-r----,..- -~ i ·,~\ ll : : 'i \ ~ ll ; ; \ · : ~ I ~ ~ · ; . ! .: ~ . · · , . ! ... .tl.-+-·· ., __ H ..... 1 ... , .. -' ..... ·t··---+·~ ...... , ....... ....•.• 'I" .. 'I . I . .. .. ' .

1. pi: I . 1 ; , i '; r' · · .. . . ' .. ; i .. :~-.II . . I . . \' . I . . . . I '·~ ',, :, . ·t" I' . ·: (~' ,._:~ ' 'I i-; • -··- · --- ---r-:-7 • -r • , :. :I· r · : t:, • • \. r::l . . . . ·... · . , · ··:

' ' . \ i ' ' ' i I l : ' 1 : ~ ' · ' .... · ; · • ' ' i ' ' ' ' ' ' J ; : ' ; ----- L~-!--•· -~-1-;l·-- _, ___ J_\·•··· ·-----f' !...!.., .. ·----!-;--~~ . :>I: i.: :; ; :I :: . :::: . \; :. . '<: T::: I;';:

• : • • ' t I • : • I I • • ' ' I I • ~ I ' I ! ' ! I ' ' .

--;-.3~1,''- ' I.'-, - - I . ! -r--.. 'I' .... I ., ...... r . , i!: l : l: I! 1: i: 1: :::. :I' . . ; ! i: ··--, '·'-·• . 'Tj'' ' ; -, . ·-•, j . •-- • ' ·- ·I •·• ·r •

I ; ; : i ~ i : i l • I l • • ; •; : • : : 1 'l : • • : ; l ; :......z-+-i+- · · · · · '- · · · · I · ·. I -· · ·

~u 1

i! i Wil" l· ! hl;: , , ? "" ,,,, [,!, ·d·" .. ,., .. [ .. '

j ~ b: i i: : rtit ~:t; ftt~L. ~: ~~tn: ~ ~-~- ~. Equal Parts 8% .At"llino Acids Soljbtion and Dextrose 2 Gms/L (right), One Hour Post - admixture.

..•

..

-...J co

f£t 0

~ p:: 0 w i=Q <1!

' .?.<=0 2.0

vlavelength (ni!_l) 2 , 1 o:: 'ti.' :7."",,.· 8sr-~+:-:-! t_ : l i ! ~; ~ l i : ! ~ t : : : i : : : : i 11 · i! i 1111

t ! I ! ! I ~ ' : 1 ~ I • I •

. . • I I ' ' I ' I : ' ' : I -~ :· : ,,. " I : " . ' I . : I ! i ' II I ! : : I I I I ' :· I ul ~-r- f-ii;;· ·~~·~,:!:: :j~' ·t:

L._L..:__ .-~-L..:.-1 -·.~ -- ~u- --t-.1 ~ ' I . I .. .. ,. .. . . . . .. .

' '' . . . i : . . . .. I\ ! . I •. . I ·. ; ·I ·. ·. ' I ' I I' :. . . l. , i :: . : i ; ': t-T--.1.__ -'-·-- -···-~.- -1.0-t--r-·

'11 J:: ,. ; j . ::· : 1: :::,,!:: rr-·v·-. ----: --- ·j··' .... ··--. ·-···j ~.-~

.±~l:~.~i+ t~rrm HH ~I\, I or .. ,, I''" .;.,t~ ·i·~-+,4-!-i!l.

. l : ; ~ r 1 ; ; 1' 1 i ; i i : ! ; ; : ~ ·1 ·:::-J::r ::l! : ,-:-: :r: h-:-: l: ::: l: . ·:::i ::-,: ~t~~~ : 7tp l · .. ··r·· .... "·' .... Hfl.;.,, ... , .... ;!~ •.. . 'I . . . " . . . " . I I . , :. . . . .. I.:. '; ... : . . .

! ; : : J ~ . .: : > . -: t i . : ' . -;6 . i ' i ;:

I ........ -"I . '·I,... l .... ,.. . . . .. I +; I :: ;j : :: l : : : ' ! -.: . . . HI I : i !---t·--·--· ·-' +--'-- -~- . ..

, .• ·. ! •. ·--c·~.,. ;:l I \ .. ,/ .. \_... ',.,

~ : .. · . · ~-· :, . .: r -: : . : : · 1 , ........ , .. ~~ . ru I : : : i : •· : ; tT·, : : ;:·· ~ ,, : i ; ~ : : ! : i -..----iL- .... ,Lj..., , ...... __ 1_ .... , .. ··-.·r .. +,·'

. • . .• . . ·I . . • . f. t' . ' I 'I • .. , .. ' { . . I ; . '. ; ! ' , :. , . .

; ~ ; i 1: ~-1· .. ; : . : ' -~ : : . ~- J•< : ; l :

-~_._L._: ~ . ..:..:....1_ :.~ :. .. .. l~ .. - ..: : : ·, ~.;.... • '..: 1 ... I' l . ' • • " . I . ' :t. l ,. • , . • . . : • . ' ' . l . I ' • ' , ~ I

1. \ , .. -~-~+-L_ -~ : ~ . .Ji~l :; I . . '·I'. . '. .. \• I: I I . :. . . . 'I I '; • 'II • - ... - i - . . • - ·- • I .. . • . ~ . . . .. ' ... . . l I

I·:: _L __ lL. _lL. : · :_ ~~-~-· ;.~ ... , ~-~.. :.j:.:· .:;: ,; .,, l!7j +~~ -: :... :·H~ !~-:-: : t! i r -,: i r: · · : ,, l h , . - . : i I : 1 ~ j ; I !" I _l\ W.Lil.J

~ max

267.5 278

Fig. 20. U.V. Spectrograms of in \vater 5096 (left) and Keflin

A max 23?.5 262.5 278 282.5

; iii iII I I j illl ; i ::! i iii; iii j i :! : ! ::: ··-'-~ ··~ ·r:! :ji: ij;:f'li: ;: 'i; r

-:-- .. ITt~~· ~:-;-:·r:-:1:, ~-- '+J..I +: ~-i ;_f~~ -:, .T:~:-; I • I I ! I : ! ; : ' ; ' • I . : ,. . ' ;

I • ·'I • • , • ' '! • '. t ' j'

II. ' ., I I"' .. 1.· .. ··J·"I ,,;--;-: :~: ::;·:1-i··· ~-::r-~·i: -~·t:;.:..~ -~:~,::~ r I I , ----;-· _:_.,; 1 r~ • -'-' ~IT' : ~-+-, ;_ __ ~~ I •1: . ' I • / I . J . : 1 1 • : I ' I I'\:' :'. /l ., J\ . :! . . I I:.:. :: .I

•·: . • ~. . - ! . . . ' ' • .., ,_; . - .•.•

I ' • 1 I I l ' ; ' : I ; . ' . . .. ; ·! . : . I " : ' : • I ,I ' ' ' : . ; '

\1.. 1 • • • . · I · · · .. · t [ : f I \ ' ' l ' ; : r l

: ' . ~[ ~ { : i i : ... · : . ', : ~ l : : : . 'j\ ; ·: ! i : : j· . ..: k. I'. 'J • ' \ ' .'.. . • . . . . '.'I

i:11 iii! ·:l!: :"fTfl~i!~ .4 - :~i: :L:t>:: 1 i l .. j. I I'll i J ; :. ; : \-:-: it;·; I i 't: !';·: i 'I • ! ' 'I ' I II '1\ l '!' .•. ' •. ; '

! i . ~ ' i : ~- : ' I ' • 1 n· ; i i : . : r, j·r ; j ; ; i . i ; ; ; . ; : : -~ -l .. l·t·•·' .j[,J~L. i .. • , .. _, ... ,L .. ......, ....... ....; •ti· .. i--"·• ~j1' +h :w :; ~~ ~:1 ~~ llk I : : ' ' : : I : '

. ll ' ··I . . ' ! ' I : . j : ' .. , ' ... , -- .... t·"· ... ... -- I -- .. 1 j : j 1 i . : : : : I ; ; j : ; ~ i : . ; ; ~ . i ; :~~ ~ : : ; ·

·...,-~-:· "-·-'·". --+··1--1-·-.. ------~ .• --"--· ----1- -·- ~-r----

1 l ! j i ( . l l: 1

j :1: : ! j j : j j i L !j ~ .;l l; li; : :·;! i II.' I'; I . Ill ,. :1' :''I ,J j' '':. ':!; '.I .• ; ; l

i ! .;;·' 1·1' \~.~ '!.1 ll'il ~:t· :t:t t!~t ~-~!!

Equal Parts 8% Amino Acids ]solution ~nd Dextrose 2 Gms/L (right) , 12 Hours :E'ost - admixture.

--.:1 ...0

ri! 0

~ p.':l_

f§ w p.':l

<1:l

1 hour

i ' ' ! I I

• j ••

. : i l ~ :J..\

~ j i". ::.,I ; : ! ~

~--

Fig. 21~ U.V~ Spectrograms in \'later 50% in combination

\'lavelength (nm)

~max 279 Shoul4er 267.5 287~5

~~~ 279 Shoulder 267.5 272.5 287.5

1 ti 3~0 ' !II ._. ; .. --,

-'-'-''-+·+"--'-'-+--1---l : ' ' l ::: L-J ! l: ~ f\! t: j: l:, :: ~, ,-.. ..~ 1 -: .-,,:: :

1,

' : i ; I' i 'I: I ' '" : I I .. • • I ; :: i I 1 ! ; ! : ! . ; i . r : : ~ : ! . , ; l ! . ; · ; ~ ~ .

: ~ 1 '· : 1 i : :I! i ' 1 , ! :: : : , l : : : : : I : ·

IiI· ''I' II!! Itt': 'I i' I ; : ·_ ; :: :: ... ; : " l.L'. II I ! ! I:': I:: '::.I::. . . ! ·I''. ·I· .. :!

H1 ~ 1!!; itl: /JL: ::)·::~~: :.j·: .. [:::1 ~- ~-,~~, -; :-r--t--,- :-·-·- -"-,---: 1--:---: t---:-_· +--r. : :: • 1 i": I ; . : ' . · · · .... :. : . i

<,.;:,~:IJ::.,·. :~---, : ; .. : :· •_:! ! .. ! : • ~: ;_; ' i ' -: c . ;_ . ; -' - -. - ~--· ,'- . :. ; : ~ ;: 'j! • \ l: 1 1 • ·, • ' f : ' I I .II '! .,. - ' . I . • • I ifi·t i ., :- l::; ·::,. . .. ; -, i . I: , . \ i! IJ 1 I I :I. I' . . . I ' .• t -'-'·-~· :. : ::'-.r:·_· __ :_:_ i--- .c_.L: ______ ...::' ! ; I llil t i i! :t;, .. : -: .1· . . , ·I' . :l I I. I, 'LL: \ t • I' I • • • ' ..• )

~-r::1 :-;I ~-,~-~-1 :-r.··--:-- -:·r--·-:- ~~--~ : ~ ; :- • i .

1

.. : . ! . • . ~ , ; : . . _ I

TIIf Trri J'-:H:-_: i ·:-··: i- ~: ·r :·:: F~ .- ~--n '!': )t.;i;: :I i:! i ;; l ... : \ : 1: . J' I: . : '•'·•hi··•' i't'•·lt•<·· ...... ,.,. ff~·-l·j---·j··-- I-I .• , i'! II . I' )' I.' ' . • . • . I I' ,., ; I' ! I' ·I : .. I '. . .. :. . . . .

).~: ~~ i:' I I:; i:.;: ':I::' :! \ ; :) :; .. : .. :; \1·!, , I;;/ r:.' :·::,I . .,., :·:--:·: L-:--. ·--: ~t

;._4 :·I I! :I i /I ':\: . ,_: < i:: . ':: :L::. . . l ji'il i!i: :f~1_-;:,;:_\ ;;:: ;. :,; ·I I' l' lil:l':i' ;•I' "'~ •. ,. i• .,·; ·-; r-1~w'--1 -t-111 i 1-h+t:i~' :-;: i1' :·t: \ ; ;~-, -~ ;~~ -:7 -l3 i!t; ~~~,;!!:;~: ~:!:··~~ :;;: ;;•; ':;;

-~-{-U~-tl1tf H~, ~h·1 ·:+F· \:!~I~- 1.::H .:..d' ,J!: 'i,: ,1·: ~.:J..;i.L' .:..::::;~_=1·.: iii! 'ill f~;,· );j:l ,,,j,"!;, ,,·,· ;.~- ;--:-~. :I I ! I t!! : ( i. : j· :! ; ' : L'. ! 'i '.:. -t -

1·-l- T -,: j·f·r·ji8" ;·;-~ :· •-;-;I '-:--;--:· -:-~..,. , ~.' --~-:-:-

i I \ i \ \ j ~ ! '. ~ ; ' : '1 i I ' • ' I ! ; ' • ,

::-1, :~~: -H1:+ ;} -.:-hf:T:'::1;::. :,·

1 l : 1 I ! I I ~ : ! ; I ~ : I ; < : ' . ; : . ; .. : : . ; . . . +-t--1--•- .. L. -t -- "1··1 _--'- - --· -- __ , ...... ~----r· . ~ --- -:!li !! i !ii\':.:: :-:··; (~:_\:~

I

of Equal Parts 8% Amino Acids Solution and Dextrose with Garamycin, 80 mg/L.

OJ 0

r::::q 0 ~ ~ gs C/2 r:q <4

~?~ 1 . . I . .~712 . ;_ -~· _, _, ~~ -L' Wavelength (nm) ~P, 1 =l"" _ _it£ __ , , __ J__ .3f0 , ' ·; :·:·· -·:! !:,T-t-:, :1:: ~~~n~ 1 ,.r:,!l 'i! ~J!l[I!ii:~~~lliLLL~_:_~-~-J :i:G~j·-·J I \::::;, :!!; i::: ili;jll!! !Iii !if: liil !iiillii; !~:li 1 ···l·!iiT!:: :1 ri i ,; : :i:rt;+;i: ;:,:,il :1 ' ll.i' :,:; :'f~,!;ii ~~~~~- ;·,.;;~:· . ; . I : . . ; . i ' I i i . ; I : i ; I I I'll i i i i I i I' I : I 'I iJ I ; l ! I i : . I i ; i . ; i .. i' : : :- . ! ; ! i I ·I·. I!,; , :I: II! i I . ; i I!. I I I :1! i! I! ; I I: II::. I:: i • , :·I .. ! :

-~ .. :._ .. !~:~? ~~"·, t • ~;r; 1 [;; 1· !i-1

:1-ji i~iil :ii: ill~;;!!'<;: ___ ;;j;,. · .. ::i 1 - : .. 1 ' .. - 1 . , ·. • • . 1· , , I : I 1 . 1 1

1 1 • 1 1• , • 1 I 1 !· I • 1 ; 1 . 1 1• ; : • : • ! . ' 1 ' • · 1 .. • • 1 : : • - - .. I J • : I • ' ' I : : . ' ' . . ! ' . I I I J i . I I . I I . i . ' ' . ~ I : ' I I : . ' I • • . • • '···'-'---'~- ·---· .,.J ..... ·-··.·. 1:1. 'l'ill'J., .. ,,, ''j l,·jl, ,1,, r''l·ri'"· ·l··j·.•: ,.,.,,. . .. 1 i . i ' • i ' ' I . : ; . ~ I . . I ' . : : : ; l I • I I I I I l : I r I ' • 1 r . ' I I I I I I J I I ' I ' . : I I ' .

i: • -; •• -;' :: .. i ' .• i;, i i '. Ill' 'I I 'I I i 'I . I i iii I'; I IiI I I I I i 'It' i ''.' ' .. : :. -....

::::::i;;_ :::;:i:~~·j;;:;l:i:: 1 ~!\:,:li'l.)\!iilli !H' >max u:.'liil! ;].,i\!:i:~ ;~![\_;:q<;:,j::-: :;.:! f1 :~~. -n- ccf> : l;mt: 'J if@T]}T ~~;ulaer 1 mtn: r: . 'lTi'!f[IT! · r~ r:rr- 171

.. 1,. ····- , :-•1·· 1,, •. ; '· .,,:l•·i'''.''! 1 1 1 2o7 5 111 1:.• .::··:•:· :··::·;· :.::·

1 :•: I·· : L.p---~-t·:-:-:- r-·:--:~1· ~~-::;-;·:·;: ; :-;--:! ;-'-;-' j :·H tj--~ ·~·i-: -~~~ 287 -5 ,~1+; ·-;~~-:- ·:i i-, r :·i-;; -:--· :~~:- ~ ·t ·-·--.'. ~-+---~, 1- . :I ! _, 1- : : : : : I : ; ; . i ; . ; !; . ! . I ; ; : ; I : i ' ! ! ; , i it' i'~ . ~ - I" : 1'1. i : j : r:-:1: i ; : : j. : ! ~ - . . · .•. ! ; . . I ... !

' - . . . . . . . I 1 ' . . ' ! ' . ' I • ! I ' l I j • ~' •. ' ! . . . ' . ; . . .. I . -L..:.. . . I

f : :; . ' . . . . . I . . . . . - . I • ' . ' . . . I I ' I l • ~ I : ' I • • ' ' •• : •.. · ••• I . . ' .. ::d~·::: _.: :::1·: ·:,:• ::1:! 11:; 1 :1!!.'!J:,!:I :J: l!!t !t.r:!::!: 'ri!!· 1 '1':.'··:~-r: j::·:t L : : ; : · : . : . : 1 : ~ : . ! : ~ : : : i : ! : ! 1 : i : i : 1 i 1 1 : : i-i · i ! i :1 i ! : ! : : : 1 : : , i 1 : : : : : :: : 1 t i ·1 > J ,,I,, , , , , , , 1 , , I 1, • , , , , •, : i , l t l • •: I :! , ·I I l , . • [ l 1 , . 1 .. ; . . . 'I\.'-. I .. ; . \\I! I! I , II'\:; I\ ! ,I :I I ·II:.:' :::I .. ! ; j·; i' i -; ! •• I\- ''

:-:-.- -:~---- - -~L --:·;-: j·--;--- ·~-~-: :- :-; r-+-t;·: : I: .. L!I ~·· ·i •i·i ",·~~ . tr·: jr 1Ti· ~~~~ 1, ... : -,1 ;· • CT"~- ;-;-cchc-r ;-' . :; :. - : ; :: ! :; :I;:: ::; i;;:: ; i;: :::: ; :-t- ~tt1:-t-':-:-f::; ::I~--.: :; ~; ~:::; ; :: . : " ' ; : . . . . ' ' . ; ' . l -. . ' I -\ ; . ' I . i : i I ' I ; I I . i ' I . I I ' I I ' '; . I . I : . ! t ' l ' , ; ' ' ! • . ' . • . . • II ~ . . , . ~··:-~:t::i: ::·:t:?:·~ :.:::,jJ~; ~J1jl~;~j ;)ri iit: ~ 1 ·\: !·t·l··J:i·J-~-f··~·-;·~+1-~:··~·:--f'~:.;.i~f_:_:,_;-. ·~·-:_~~-: ~---f ·····•·.'· ····I···· ··i· \ill ;,f:l:i'' ''I ·1·'8'' !JI: 11'1 ,J;,J• .•. i I 'til ,, L ::1'" ;. I ::: :.;~:; ! . ::~- ~:::: :::·''~:: ·:\!El .!-~ :·:: -~:: !~~~~~f~::.:-~-H-:-;--f-1 ··: i'•• P···· ····r~-, ··1:1····. ,,! :' 'II -!1' :,·: ,;!: ••.. , .. ," •. , .. ,: --:~:..1

~-~~"' :': !;Y~-:U·t: ~.:,ilL,::!!:':· ,~~~r :rr:;~,r;·: -:m:{': ',ll\'~'m. I;--~ ___ .. ,, .. ,.. .. .. ,~'Lc ..... . ,..,_-... ,.,:Y .... ~-.·--:-·j-•-- -'.i~ij·'+•·· .. ~!.should r __ , . -,- :+-l·:f--"-- -., .. r~1-- 1 ·--- +---.,-.,.~-- ---···--· . . . ; : ·.j: r:: : ~ j : 1:;: · ; 1 • ! i!; r · ; i 1 i ! 1 ~ ~ ; :; e • :! : \ 1 . . ·;,: ; t 1 ~ ~ · •· : i i;: = i.: :; : · t

, i· .. ·!;-' .. '·''I' ... ; 'II'::,' :JI. •; 267 5 I l ,,. '! /' ;' '·:''1 ·1· ... ,,, .. :·'j : . ;·t:,: .. :, :::: ; :: · :-r· : i;

1:::: ':·I . ; I;++ • :1 ·: ,\;:. , :·v-f-tlli·;:. /1:: .•· . ; :1 i;, ; : · >

''I'll•' .... 1 li· ;,.1.,! ,r! ,I•, !/1 l•'i ,,1287.5 111·1'\:1 1!'i ;.\ · :ti:I',J ·:;TI, r·:· '"I~ -:-~·~- ~~{+~~-~ 0.·~-:--:·-i-- -~-~-!-.. ,·~+·!~ .. ·~-~-i· -;-+~~- ·~-~ 11~ i- · \-;-: • ~r- -r+r ··7 r~-. ~-11 : ii ... rl:-:· :-:-0-.. ;.j..

1 .. , ;" ·::·j· :L ::;; ·•: .\:: •1 ,:· :·1 jjl 1

1 '" .f, j''!l, •il''1l'\t· ij ·!;. :i" 1 • 't . . I . I • I ' • • : '\. • ' I I ' I I ' . I I • I I l ; t I { . . . . . I ' ' • . I : jl • . • . ' .

; .. : . . .. . (. : . . . : : ; . . ~ ; : . t : ~ : . : : : ; I . : :.3 ; ; . : ·~- ~-=· ; I : ~ : i : ; ; ~ : : ; ·. . : : ; I ~ ' ; ! ~ . ' • . . I - I . - . ' . 'l" ; . I . : .. I ; i " ' ·, ! I . i ' I ' I I I II ! II : ;; I ; ! ii.. : I ., I I . ~ ... : ' i ' !I \ ' ' j : ! ' .. I .. '--. --- . : .. : . . ! : ~ ~ ~ ; : : ' ; I ~ ! ~ : ' l I ' : ' i ! I I : I i 1 . ~ l : I t I : • 1 . : ! : j • . ; . I f l : i i . : I i . --~---1----------·--i---c-- -.---'---·----r-;-n···- H-~~~- --rr ~·lt ,, . -~~~ .. ,.,, .. ~-~~~~··-·· ····r-·-'-- ..... I . I . • . " : : : I : ! : - ' . . . I . ' . , ' ; I ' I ' I ' • . i ' I ' .. I . . '·' " ' • ' , ,. • • ' .:.·,~_, .. ;;. -•~·''f!lil!ij'•:'!! liji;l!lj··l 1 j•l''ji~'''j··

,-p·l-~~~4~-l- iJ- ~Hl1t -rt~ : i ill iil~!~l, +~iW ~'IT~~ 7h~ m -..... l'. 'il'·l'd'' ·r,,' I ; ! ! ': ;l·j:' ·-:.!•··!. 'II•! ·' - I·- . ; ; . • . . . l .• I . '.. • ' I; I': I, : ' I '' I : !·: •. ' II:. . I ~--t-::-· ·+:'--;; .: .. '+'1--::: ·iii· ' ·t r[·ll+-t+ ~ri+·--' :.;..:Ji·;:-;-: ~--:..;-r-:-~ -~--,

t-. · ~ · :I: ;. ::- ·:' : i k_iL.!; JJJ ~ 1 \: i \:: A i I; : · ; : r' 1;:;: :,:\I i :-: -. ·: l

1 hour

Fig. 22. UoV. Sp~ctrogram~ in \'later 50% in combination

"" "' 1 P ' · Bo! Am. O.L ..... qua ar-cs ;o ~no

with Kantrex, 500 mg/L.

12 hours

Acids So]ution and Dextrose

'; ..

co 1-'

82

Thin Laye~ Chromatography

Duplicate samples of the amino acids/dextrose solution

were employed in the development of a standard chromatogram.

Utilization of 0.5 microliter· required 7.5 hours for develop~

ment in Solvent System I, and 3.5 hours in Solvent System II.

Having established a standard chromatogram (Fig. 23)

duplicate samples of the hyperalimentation solution were sub

jected to admixtures of 11usual 11 therapeutic levels of electro-

lytes and vitamins. Similarly, 0.5 microliter was transferred

from the samples 1 and 12 hours after admixture and spotted on

separate plates. Follo\IJing development and visualization, the

resultant chromatograms were compared to the standard.

The comparative chromatograms on Fig. 24 may be sug

gestive of chemical interaction at the termination of the 12

hour period of investigation. One component of the chromato

gram vanished; another exhibited a loss of spatial continuity.

Conversely, the spatial arrangement of the chromatographic

components of Fig. 25, appeared essentially unchanged. This

latter graphic representation was suggestive of chemical

compatibility ..

~--~-~-- --·- --------- -- . -·------- -- ~ - -- --- -- -- ------- - -- --- ----------------

83

-

-·

. -.

Solvent Front, lst Dimension

I

s:l oo 0 ·ri til 0 ·(\n s:l 0 Q)

s vv-•rl A 0 rd . s:l 00 (\J 0 .. 0 ~ 0 0 ~ D f.XI

~ ·-(!)

I>

000 1 r-1 0

(.!).

o~igin .J ~

Fig. 23. Standard Chromatogram of 896 Amino Acids Solution and Dextrose in \'later 505'6 (1:1) - '

-

.. I

I. I

I

Solvent Front, 1st Dimension

. I .. Solvent ~.,ront 2 1st Dimension

.s:: 0

·rll 0{9) w s:: (l)

8 ©1 @0" OJ ·r-l ~ ( )i ) A

rd (} s::

C\.1

00° ({)) r-

rf ~ 0 t H

h lil

4-' s:: ro (1) Q)t_l. :> ,...; .o U) origin

s:: 0

•r-l ro

~r s:: ( J(\ . (])

~ r5Q c~) s ·rl . 'l

R 1_,

rd 0 ' . s:: C\.1

~, 'b ~ ... oOb ' 4-' s::

rt/ 0 .

H I lil

~ (J)

"~m I> r-l . ~) 0 m origin

I ---------- I -- -- ---~-------------~

1 hour . · I 12 hours Fig., 24. Representative C:b..romatogram o:f 8% Amino Acids. Solution and Dextrose in. Water 50% (1: 1) Combined "~:lith Potassium 'Phosphate 20 mEq/L, C!3.lcium Gluconate 10 mEq/L, Solu B Forte .10 ml/L, Folvite 5 mg/L, Rubramin PC 1 mgJIL, and Aquamephyton 10 mg/L. ·

--- Standard Solution --- Sample Solution

I I i

I

!

:

I

I I

,. . .. . ': ~ ..

~. f ·~~

~

;

:

Solvent Front, 1st Dimension Sqlvent Front, 1st Dimension

!=1 s:: 0 .. 0

•r-!

OQt' ·n

U2 U2

~O,"Ii s:: s:: (J) ~,.) . Q) 'u -s OG ttJY 0

s o o/J-, -~ ·rl •r-1 A 8 0 \_)'-" \ ...

rd OJ 11 s:: ' (\J

OJ (\J 0 0 ")

• ((]; r:l ()!{) .. ,; ~0 0 n\ ('). -!..::> v t:l s::

0 0

& H A

-!..::> r&Q -g s:: .. Q) (J)

:> (Y\J~ :> ,..., ,..., 0- 0

Cf.l origin UJ

1 hour Fig. 25. Representative Chromatogram of 8% Amino \'later 5096 (1 :1) Combined with Potassium Phosphate mEq/L, NVI 10 ml/L,_Folvite 5 mg/L, Rubramin PC 1 --- Standard Solution --- Sample Solution

\) ,J ~tJ

\./

RfB ',./ .

origin I

·I 12 hours Acids Solut:ion and Dextrose in. 20 mEq/L, d(alcium Gluconate 10 mg/L, and l,lquamephyton 10 mg/L.

..

(X) \J1

I

I

I

I . I

86

Results of Mi.crobiological Assa;y

An evaluation was undertaken to ascertain the degree of

degradation or interaction, if present, of selected antibiotics

admixed separately v'lith: a) the· amino acids solution and £.) the

amino acids solution in combination 'u'Ji th therapeutic concentra

tions of electrolytes. Potassium phosphate, 20 mEq/L, and

calcium gluconate, 10 mEq/L, constituted the additives in

section (b). Any significant decrease in potency 'I!Jould mani-

fest itself as a reduction in size of the zone of inhibition as

compared to the control.

Polycillin N displayed a dimunition in its zones of

inhibition relative to its aqueous control in both section (a)

ru1d (b). Refer to Tables 19 and 20. The activity of Keflin

appeared to be en_~anced in the presence of either solution

(Tables 21 and 22)c The efficacy of Kantrex seemed unaltered,

if not enhanced, in each set of sample solutions (Tables 23 and

24). Garamycin maintained its activity in combination ~:lith the

876" amino acids/dextrose solution alone and inconbination \·lith

electrol~tes (Tables 25 and 26).

§tatisj:;_ical Evaluation.. The variance of the observed

data from the microbiological assay indicated a statistical

analysis ·was requi:eed. to evaluate the significance of the

altered zones of inhibition. Consequently, a "Student t" test

was perfol ... med. Tabulation of these calculations are found on

~['able 2? ~

The efficacy of Polycillin N, Keflin, and Kantrex in

87

the amino acids/dextrose' solution, -with or without electrolytes,

did not appear to be significantly different from control

samples prepared in sterile water -(o.2;,P<O.l, O.Ol>P<O.OOl;

0.2>P<O.l, O.l)P<0.05; 0.5~P<0.3, 0.5>P<0.3). It appeared

that Garamycin might be significantly more effective in both

hyperalimcntation mixtures than in .sterile water (P<O.OOl,

P<O.OOl). Hov1ever, further investigation should be completed

to evaluate this statistical significance.

Table 18 ·

The Inhibition of Gro'."rGh of Stanhlococcus aure~ts by Polycillin N at a Concentro..tion of 20 }{g/ml in.. a Ni:xture of

Equal Parts 8% iunino Acids Solution and. Dextrose in lvlater 50%

-----------------------------------"\:------Zones of Inhibition (mm)

1 hour I 12 hours

~~tibiotic in Ster. Hater for Injection

Antibiotic in P~ino Acids Solution

Antibiotic ·in Ste:r. Antibiotic in Amino \rJater for Inject~1 on Acids Solution

27.3 27.0 27.2 27.9 27.9 26 .. 7 27.3 28 .. 1

27.7 26.2

28 .. 3 25 .. 9 26.3

27.7 27.5 27.2 26 .. 2 27.3 26.8 27.5 27 .. 9 27 .. 6 28.1 ---- 25.6

g6.0

He an 27.53 +0.355 26 .. 93 +0.877

. ,-

26.8 27.4 27.2 27.6 27.1 28.0 28.0 25.6 26.1 27.2 26.8 27.2 27.0 27.2 22 .. 2

27. 09 +0. :624·

25.7 25.4 26.1 27.0 25.4 25.3 2 r- 1J.. :::>. . 25.8 24.6 26.0 26.3 26.3 26.0 25.5 26.4

25.81 +0.582

co co

Table 19 ..

The Inhibition of Gro"t"rth of Staphlococcus aure~ls by Pol:yvcillin N at a Concentration of 20 ~g. ml in

Therapeutic Concentrations of :raectrolytes Admixed to Equal Parts 856 .Amino Acids Solut;ion· and Dextrose in vla er 50%

Zones of Inhibition (mm) -112 1 hour hours

Antibiotic in Ster. Water for Injection

Antibiotic in Amino Acids Solution ·v.rith

Electrolytes

Antibiotic in stJr. \'later for Injectilon

Antibiotic in Amino Acids Solution 1:;i th

Electrolytes

He an

27.4 28.2 28.2 27 .. 7 27.4-28.5 28.5 27.9 27~1 27-5 27 .. 0 28 .. 1 27 .. 4 27.1 gz.4

27.69 +0.506

28.2 27.0 26 .. 8 27.2 27.9 27 .. 8· ')n /'" c...().O

26 .. 9 27.6 27 .. 2 27.4 29.4 27. Lf. 26.8 28 .. 3

27.63 +0.743

25.5 26.6 26.5 26.9 26.5 26.6 26.0 26.4 25.5 25.7 25.8 25 .. 2 26.4 25.9 26.2

- I 26.11 +0.·14-98

25 .• 6 25.7 24.2 25.1 25.5 25.9 25.4 25.9 2L~.3 25.0 24.3 24.5 24.5 24.6 24.8

25.00 +0.610 co \..0

Table 20

The Inhibition of Growth of Staphlococcus aurev~s by Keflin at a Concentration of 50 Ag/ml in a Mixture of

Equal Parts 896 A.rnino Acids Solution and Dextrose in lvJater 50%

Zones of Inhibition (mm) 1 hou~ 12 hours

Antibiotic in stJir. Antibiotic in Amino Water for Injection Acids Solution

Antibiotic in SterQ Antibiotic in Amino Water for Injection Acids Solution

33.2 35.0 35.4 35.8 34.8 33.9 34.2 34.7 3'' D ...... 35.6 34.2 35.8 33.6 35.6 34-.1 35.8 35 .. 2 33.4 34.6 35.0 32.6 36.0 33.0 33.8 3L~.2 31.2 33.9 34.2 33 .. 4 35.8 35.0 35.2

35.2 35.0 ·35.8 31 .. 6 34.4 36.0 34.5 33.6 35.0 35.4 35.0 32.8 32.6 3L~.o 35.8 32 .. 4 33.0 3L~.o 36.0 3LI-.A 34.1 35.0 32.7 ,36.0 ~ .2.2&

IvJ:ean 33.60 +1.204 34.47 +1.4-12 34.58 +0.i827 35.15 +0.6?8

'.0 0

Table.21

The Inhibition of Gro,·rth of Staphlococcus aureti.s by Keflin at a Concentration of 50 ,u.g/ml in Thera~~eutic

Concentrations of Electrolytes ACL.'Ilixed to Equ.al Parts 8% Amino Acids Solution and Dextrose in lVa~er 50%

i

Zones of Inhibition (mm)

1 hour 12 hours

JL~tibiotic in Ster. Water for Injection

34 .. 1 34 .. 0 32 .. 7 33.9 32.8 35.2 3L~.2 35.8 32 .. 3 34"0 35.2 26.7 32.8 34.2 32-2

He an 33-37 +2.122

.A..~tibiotic in Amino Acids Solution \·ri th

Electrolytes

33.9 33.2 35.0 33.7' 7,7, 7 ----· 32.8 35.0 33 .. 2 32 .. 8 35 .. 8 3L~.o

32.0 35.0 3L! .• 7 36.0

34.05 +1.156

Antibiotic in Ste\r. i:Jater for Injection ·

34.2 35.2 35.8 3L~.l 34.2 35.2 )4.L~ 34.,4 33.4 35.6 35.4 35.8 35.4 35.6 34.9

34.90 +0. 117391 . - '

Antibiotic in k'Ilino Acids Solution -v;ith

Electrolytes

34.9 35.8 35.9 35.4 35.2 36.0 35.8 35.8 35.8 35.4 35.4 34.4 33.8 35.3 2h0 35.26 +0.696 '.{)

l-'

Table 22 ,.

The Inhibition of Grm~th of Staphlococcus aureu!s by Kantre:x at a Concentration of l.J-00 }tg/m1 in a· Mixtju.re of

Equal Parts 8% .ibnino Acids Solution and Dextrose· in il:later 5076

Zones of IrJiibition (~~)

1 hour 12 hours

Antibiotic in Ster. Water for Injection

Antibiotic in Amino Acids Solution

Antibiotic in Stelr. Antibiotic in Amino \!later for Injecti:on Acids Solution

He an

15.6 ]_6 .. 4 19.5 18.Lf-16.4 15.4 19o0 15.8 18.8 1? .. 2 20 .. 0 17 .. 0 15.8 19 .. 4-. 16.3

17.40 +1.52

18.7 17.2 19.7 18.0 16.7 17.6 20 .. 5 19.2 16.L~

18.8 18.6 16 .. 6 19.8 19 .. 5 18.6

18.39 +1.27

16.6 17.4 16.8 15.6 17.6 17 .. 4 17.9 16.8 16.8 17.1 16.4 17.2 16.5 17 .. 2 15.9

I 16.88 +0.$25

17.6 17.6 17.6 17.7 17.7 18.0 17.5 17.1 17.1 17.8 15.6 16.8 16.4 17.2 17.0

17.24 ..:!;.0.624

'iB

Table 23

The Inhibition of Gro'l:rth of Staphlococcus aurev.s by Kantrex at a Concentration of 400 Mg/ml in. Therqpeutic

Concentrations of Electrolytes Admixed to Equal Parts 896 Amino Acids Solution and Dextrose in vldter 5096

Zones of Inhibition (mm) 1 hour 12 hours

Antibiotic in Ster. \vater for Injection

Antibiotic in Amino Acids Solution "~:Jith

Electrolytes

Antibiotic in Ste~1r. \'later for Injection Antibiotic in Amino Acids Solution \vith

Electrolytes

f1ean

17.0 18.6 16.4 19 .. 8 16.5. 16.3 18.6 17.2 18.9 18.6 18.4

·17.7 18 .. 2' 1?.0 15.8

17.66 +1.163

17.0 18.3 19.1 18.7 17.6 17.8 17.2 16 .. 6 19.2 17 .L~ 17.5 17 .. 3 15*9 17 .. 2 16.4

17 .. 54 +0.95

18.4 17.2 17.7 17.1 17.0 17.2

15.8 17.2 17.1 17.2 17.4 16.7 16.9 l7.L~

17.16 +0.1362

17.1 17.2 17.2 17.4 17.0 15.9 17.0 17.6 17.6 17.3 17.9 16.9 17.6 16.7 1h.2. 17.20 +0.492 \..0

\)J

Table} 24

The Inhibition of Gro~:Jth of' Stanh1ococcus aure~1s by Garamycin at a Concentration of80 ~glm1 in a Hlliure of

Equal Parts 896 Jlu11ino Acio.s Solution and Dextrose in I \later 50%


1 hour 12 hours

Antibiotic in Ster. ·water for Injection

Antibiotic in Amino Acids Solution

Antibiotic in stJ;r. Antibiotic in Amino I.!Jater for Inject~Lon Acids Solution

17.2 16.6 16.0 16.4 15.0 17.0 16.2 16.0 17.6 17.2 15.4 17.0 17.7 16.3 15.0 ,16.8 16.7 17 .. 0 17.3 16.2 17 .. 4- 16.4· 15.5 17.1 16.6 16.6 15.3 17.3 15.3 19 .. 2 14 .. 8 15.8 15 .. 0 17 .. 1 14.6 16.5 19 .. 2 16.7 14-.5 16.0 16.2 17.0 17.4 16.4 14-.9 16.5 15.9 16.4-15.7 16 .. 3 17.4 16.0 19.6 "20 .. 1 15.9 16.2 14 .. 4- 18 • .2 1;2.2 16 .. 2

Mean 16.57 +1.569 17.25 +1. J.ll-8 15.76 +0~970 16.42 +0.446

'-!:> ~

Table 25:

The Inhibition of Grovvth of Stanhlococcus au.ret;ts by Garamycin at a Concentration of 80 Bg/ml in Ther.SLpeutic

Concentrations of Electrolytes Admixed to· Ea~tal Parts 896 Amino Aeids Solution. and Dextrose in \•la~:er 5096


1 hour 12 hours

PIGibiotic in Ster. Water for Injection

14.8 15 .. 7 15.0 14.9 lL~ .. l 14 .. 7 18.2 1 'A 0 ·'-./ .. 7

lLJ-.L~

15.0 15.5 15.L~ 11-l-.3 16.4 1L~.6

Iviean 15.13 +1 .. 07

Antibiotic in Amino Acids Solution vJith

Electrolytes

17.2 16 .. 6 16.4 17u5 16.6 16.6 16.,6 16.4 16.7 16 .. 8 16.7 17.4-17.1 16.7 1'7 .. 1 .;:;;;.L.._

16.83 +0.347 .-

Antibiotic in Ste~r. \rJater for Inject~on

I

15.7 14.9 17.1 15.0 15.0 14-.7 15e5 16.2 15.0 15.5 15.5 16.0 15.5 15.2 1S.4 .............._

15.4-8 +0.1608

Antibiotic in Amino Acids Solution vdth

Electrolytes

16.5 17.3 16.6 16.4 17.0 16.8 16.9 17.1 16 .. 7 15.8 16.5 16.6 16.4 ' 17.2

'16.2

16~70 +0 .. 652 '.0 \Jl

96

Table 26

Calculated Results of "Student t" Evaluation

Polycillin N in Equal Parts 876 Amino Acids Solution and Dextrose in Water 50%

1 hour 12 hours

t = 0.198 t = 1.490

( 0. 9 > P<O. 7) (0.2->P<O.l)

df - 38 df = 43

PolycilJ.in N in Equal Parts 8% Amino Acids Solution and Dextrose in Water 5096 and :Slectrolytes

Kef1in in Equal Parts 896. Amiri.o Acids Solution and Dextrose in \'later 5096

1 hour 12 hours

t = 1.767 t = 1.332

(O.l~P<0 .. 05) (0.2>P<O.l)

df = l~2 df = 42

Keflin in Equal Parts 896 1\.mino Acids Solution and Dextrose in Water 50?6 and Electrol:'"··+;-es

1 hour 12 hours

t = 0.015 t = 1.932

(0 r· p· 0 ·z' "/1>. < -· ":J .J (O.l>P<-0 .. 05)

df = L~2 df = 1+3

Kantrex in Equal Parts 896 Amino Acid:;:: Solution and Dextrose in \'later 5096

1 hour 12 hours

t = 2.716 t = 0.995

(O.Ol>P<O .. OOl) (0.5>P<0 .. 3)

df = Lij elf = 42

Kantrex in Equal :Parts 8]6 Amino Acids Solution and Dextrose in \Vater 5696 and Electrol::-'~-~s

l hour 12 hour·s

t = 0.035 t = 0 .. 860

(P<:0.9) (0.,5>P<0 .. 3)

df = L~3 df = '+2

Garamycin in Equal Parts 896 _1\..rnino Acids Solution and Dextrose in Water 50)';.;·

1 hour 12 hou.rs

t = 0.325 t ""' 3.556

(Oo9>P<0 .. 7) (P<O.OOl)

a.r == 43 elf = L!-3

Garamycin :Ln I·~qual Parts B96 Amino Acids Solution and Dextrose i:n \'later 50Jb and EJ.ectroly";es

1 hour 12 hours

t ::: 2 .. 562 t -· 4 .. B90

( 0 p O~~:>P<O., 01) (~<OoOOl)

d.f _., 43 df ·- Lfj

~------·~---------~~~----··---------

Chapter 4

DISCUSSION

f-1ost often, the pH of parenterals ivill determine

admixture compatibility. Parker (90) has implied that pH,

gas content, solvent type, and moisture is carefully con-

trolled in many parenterals. Consequently, when " ••• such

sensitive drugs are diluted to larger volumes by an aqueous

solution of a different pH containing atmospheric oxygen

-\"rhich might catalyze or initiate a reaction, decomposition

is possible 11 (90). The combination of preparations ivith

~brerse hycLr.ogen ion content predisposes the miA.'ture to incom

patibility, \vhether visual or unseen. The apparent physical

compatibility in these instances should not be construed as

an indication of chemical compatibility. There exists the

possibility of degradation products possessing a certain

degree of solubility before the point of solution saturation

is surpassed, which \·Jill result in a visual manifestation of

incompatibility (90) ..

The amino acids/dextrose solution v1as found to have

a pH of 6.15. The addition of various levels of electrolytes

produced slight alteration in this value (Tables 5 to 7). The

direct admixture of potassium phosphate and calcium gluconate

in an aqueous medium resulted in immediate precipitation.

~'his finding coincides vJ'ith a previous study (66). Yet, the

97

98

hyperalimentation solution accommodated elevated levels of

these electrolytes before particulate matter appeared. Quite

interestingly, magnesium sulfate seemed unreactive in the

presence of either electrolyte, contrary to earlier reports

(64,66;68,91). The reason for this remains unclear. Perhaps

'the complex nature of the amino acids/dextrose solution pro

vides physical barriers to possible interaction between the

electrolyte additives.

The order of electrolyte admixture VJas determined

pertinent to the compatibility of elevated concentrations ..

Potassium phosphate should precede the addition of calcium

gluconate. This would be in agreement with the findings of

q?llin, et al. (86). The amino a~ids/dextrose solution vias

also found to accommodate 100 mEq/L of either electrolyte as

a single additive without demonstrating physical incompati

bility.

The addition of Solu B Forte to the basic stock solu

tion \vith electrolytes appeared to overtax the buffering

capacity of the nutrient solution (Table 8). Solu B Forte

measured a pH of 3.0 prior to admixture. The alteration of

pH b~y f1VI addition v1as ascertained insignificant (Table 8).,

Similarly, single admixtures of Folvite, Rubramin PC, and

Aquamephyton to the hyperalimentation infusate ·with electro

lytes failed to demonstrate any interaction (Tables 9 and. 10) ..

Folic acid_, with a pH of 8.0 to 11.0 (92), failed to precipi

tate in the presence of calcium salts as is reported (93).,

Rubramin PC did not exhibit visual signs of decomposition

'

which is reported to occur in the presence of elevated

dextrose levels (64,79,93).

99

The examination of the hyperalimentation solution

containing electrolytes and vitamin combinations also failed

to manifest evidence of physical incompatibility (Table 11).

·The literature has reported instances .of decomposition with

many such mixtures presently formulated for total parenteral

nutrition~ Ascorbic acid has been implicated in the degrada-

tion of cyanocobalamin (64,79,82,91,94) and phytonadione (64,

91). The decomposition of cyanocobalamin and phytonadione in

the presence of each other is also noted (64,79,91,94). Lack

of physical demonstration of this occurrence enforces the con

cept that physical compatibility is not necessarily indicative

of chemical compatibility.

The addition of Iletin U-40 (Table 12) did not appear

to alter the pH or compatibility characteristics of the amino

acids/dextrose solution significantly. T1rJO instances of pre

cipitation \':Tere noted at the end of the 2L~ hour period of

observation; ho·,·Jever, the reason for this remains unclear ..

The addition of various concentrations of potassium

phosphate and calcium gluconate to the basic stock solution

prior to antibiotic admixture demonstrated a detrimental effect

on the compatibility and stability of these agents (Table 15) ..

Elevated levels of electrolytes caused precipitation in most

mixtures by the end of 12 hours after admiA~ure. Polycillin N

appeared to be; the least stable.. Keflin \·Jas observed to be

the most stable. These data vJould suggest that Polycillin N

100

should not be admixed with other medicaments as previously.

advised. Keflin has been cited to be incompatible \•Tith .

calcium gluconate (64,65,94). This 1'1ould appear substantiated

in the presence of elevated levels of the electrolyte admixed

to the hyperalimentation solution. Reported interaction of·

· Kantrex '''ith dextrose (64,91), accompanied with elevated

concentrations of electrolytes could be implicated ·t,~Tith its

precipitation in approximately eight hours (Table 15).

To further challenge the amino acids/dextrose/

electrolyte/ru."ltibiotic mixture, supplemental vitamins were

admixed for observation of possible physical incompatibility

(Tables 16 and 17). The acidic nature of Solu B Forte appeared

~~ reduce instances of precipitat~on at elevated electrolyte

concentrations. Again, Polycillin H appeared the least stable ..

The utilization of ~NI did not suppress the final pH of the

mixture as did the Solu B Forte. Consequently, Polycillin N

stability increased, 'li'lhereas, the stability. of Kant rex; Keflin,

and Garamycin \'Jas noticeably decreased.. The reported possi

bility of decreased antibiotic efficacy in the presence of

riboflavin (65), nor the inactivation of Keflin in the presence

of Vitamin B-complex and C (64) should not be overlooked.

As this study vras undertaken to establish the compati

bility and stability of the solution of amino acids and dextrose

when mixed I.·Jith commonly employed supplements, compatibility

bet\•leen these supplemental agents themselves v1as not investi

gated. As a result, the amino acids/dextrose solution under

Hent admixtures of each adjunct and was examined by means of

101

u.,v. spectroscopy. Where possible, both components \llere

scanned in the mixture. Signi~icant alterations of the Amax'

wavelength of absorbance, and/or appearance of new peaks was

considered suggestive of chemical interaction.

The u.v. spectrograms obtained for the aliquot dilu

·tions of the amino acids/dextrose solution reflect the

phyenlalanine and tryptophan (95,96). ·The most notev10rthy

alterations of U. V. spectra occurred 't•Thile scanning combina-

tions o~ the hyperal.imentation solution and each of the

multiple vitamin preparations (Figs. 7 to 13). These spectro

grams were suggestive of interaction. Parker reports that

thiamine is readily susceptable to cleavage by sodium bisulfite

(90). The percentage of decomposition of B1 has been estimated

to be 10% in 2 hours, and 2576 in 6 hours. The 89~ .Amino Acids

Solution contains potassium metabisulfite, 100 mg/L, and may

interact in similar manner v.Ji th the thiamine.

Unfortunately, the therapeutic levels of Folvite,

Rubramin PC, and Aquameph;yton 1:Tere too dilute to substantiate

the various claims regarding their inactivation in the presence

of each other. This study vms beyond the scope o~ this investi

gation.

U.V. spectroscopic examination of antibiotic/amino

acids/dextrose mixtures proved inconclusive (Figs .. 18 to 22).

The only altered spectrogram \·ms exhibited by the Polycillin N/

amino aci.<1s solution mixture (Fig. 18).. A secondary peak at

221.1. nm appeared \·Then the solution was scanned one hour after

admixture. This peak remained unchanged throughout the 12

102

hour period of examination. The addition of Keflin to the

amino acids/dextrose solution appeared to reflect a ~ypo

chromic shift in absorbance of the amino acids solution spectra.

However, detrimental interaction betv1een Keflin and the solu

tion of amino acids and dextrose vms not suggested by the

·microbiological assay.

It should be emphasized that throughout the u.v. spectro

scopic study, aqueous dilutions were required to provide optimum

concentrations for scanning. There exists the possibility this

procedure could have initiated or catalyzed the reactions

revealed through the altered U.V. spectrograms.

Thin layer chromatography suggested the possibility of

chemical reaction upon the addition of electrolytes and vita

mins to the amino acids solution (Fig. 24). The chromatogram

produced from a sample 12 hours after its mixture presents

striking inconsistencies in the amino acids migration pattern~

Further study should be undertaken before Solu B Forte is

rated completely compatible in total parenteral nutrition. The

pictorial results in Figure 25 lack conclusive evidence sug

gesting chemical interaction5 Even though some amino acid

components of the chromatogram are not congruous,- the spatial

arrangement remains proportional. Due to the complexity of

the solvent systems, the investigator elected. to retain the

solvents utilized in the development of the one hour sample

for the sample withdrm·m 12 hours after admixture. Prepara

tion of nevi solvents could unnecessarily give rise to new

variables.

103

Initial examination of the antibiotic/amino. acicls

solution combinations using U. V. spectrosc6py \•las unremark

able (Figs. 18 to 22). Subsequent study of these solutions

by microbiological assay suggested no significant detrimental

chemical interaction. Btatistical analysis of these results

(Tables 19 to 26! indicated no significant decrease in anti

microbial effect between the aqueous controls ana. the test

sample solutions. The presence of the electrolytes appeared

to enhance the potency of Garamycin significantly. This

observation ".-Jill require further investigation. Elevated

levels of dextrose and electrolytes did not appear to impede

the activity of the antibiotics significantly as has been

reported (61+,65,91, 94). Vitamins were not included in the

mixtures,·as riboflavin is reported to cause rapid degradation

of these agents (65). It vms also noted that undiluted amino

acids/dextrose solution, used as a control, exhibited an

inhibitory effect upon the grm:Jth of the organism utilized.

'l'hese data \vould suggest that potassium phosphate and

calcium gluconate do present compatibility problems in levels

exceed.ing certain therapeutic concentrations.. Their order of

admixture also determined. the maximum compatible levels. The

important rolE-) of pH in parenteral admixtures \oJas best exempli

fied with the addition of multiple vitamin preparations to the

hyperalimentation solution. Liquid mix:tures which are observed

as physically compatible may in fact be undergoing chemical

interaction or degradation. It is recommended that employment

of Solu B Forte, ar; a routine vitamin ·supplement in total

104

parenteral nutrition, be approached .cautiously until further

conclusive studies are completed. A study of whether the

nature of degradation products are possibly detrimental to

the patient should be paramount. On the other hand, results

of TLC analysis would suggest that rwr may be better suited·

'for vitamin supplementation in this hyperalimentation infusate •

. The remaining vitamins , i.e~ , Fol vi t e, Rubramin PC, and

Aquamephyton failed to demonstrate physical incompatibility

upon ad.rnixture, as well as manifest signs of chemical inter

action \·rith the amino acids solution through U. V. spectroscopic

analysis.

Therapeutic levels of antibiotics appear to remain

stable and efficacious for 12 hours after admixture to the

hyperalimentation infusate, except in the presence of elevated

concentrations o.f electrolytes and HVI.

Chapter 5

CONCLUSION

The patient criteria for initiation of total parenteral

nutrition have been discussed. The nutritional requirements

for protein administered in combination "~.'lith various supple-

mental agents has been vJell documented. The advantages of

utilizing a pure protein source, i.e., crystalline or synthetic

amino acids., \vere described. In order to establish compati

bility and stability characteristics of a mixture of equal

P?-rts 896 Amino Acids Solution and .Dextrose in \vater 507'6, the

solution uncler\'Jent various adrni::-rtures of commonly employed

electrol;ytes, vitamins, insulin, and selected antibiotics ..

The nutrient solution appeared to accommodate levels

of potassium phosphate, calcium gluconate, and mag11esium sul

fate i·..rell in excess of "usual" therapeutic concentrations.

Similarly, commonly employed vitamin supplements and insulin

were examined in combination i:Jith the hyperalimentation solu-.

tion and failed to demonstrate signs of physical ·incompatibility.

Howcnrer, instrumental analysis, i.e., U.V .. spectroscopy and

thin-J.e.yer chromatography, suggested, in fact, there may have

been chemical interaction of Solu B Forte and NVI once admixed

vJith the amino acids/dextrose soluti9n. Yet, results of thin

layer chromatography did not appear to substantiate chemical

interaction bet"tveen MVI and the amino acids solution.. The

105

106

investigator contends further study ·Of multiple vitamin

infusion stability be completed before definite conclusions

are formulated regarding their compatibility in this nutri

tional infusate.

Therapeutic levels of selected antibiotics, i.e.,

Polycillin N, Kantrex, Keflin, and Garamycin, were examined

separately in combination with the amino acids/dextrose solu

tion alone, i.·Jith electrolytes, and with electrolytes and vita-

mins. Elevated concentrations of potassium phosphate and

calcium· gluconate in combination vlith MVI appeared to accelerate

the signs of physical incompatibility. Subsequently, these

anti biotic/amino acids solution mb,.rtures were examined uti liz-·

~~g U.V. spectroscopy. These datq suggest an absence of chemical

intera.ction. Further information from· a microbiological assay

-indicated that admixture of antibiotics in these hyperalimenta

tion solution mixtures did not significantly reduce the anti

microbial activity of these agents.

--~~---~~---

1.

2.

BIBLIOGRAPHY

Heird, \v. c., Driscoll, J. N., Schullinger, J. N. et al.: Intravenous Alimentation in Pediatric Patients, J: Fed. 80(3):351-72 (Mar.) 1972.

11eng, H •. c.: Use of Fat Emulsions in Parenteral Nutrition, Drug Intell. Clin. Pharm. 6:321~330 (Sept.) 1972. ,

3 .. Geyer, R.: Parenteral Nutrition, Phys:i.ol. Rev. 40:150-86 {j--~~~~~~~~(.Ma~.--)~l-960-. ·

4.

5 ..

6.

7.

8.

9.

10.

11..

12.

Dudrick, s. J. and Wilmore, D. H.: Long-Term Parenteral Feeding, Hosp. Prac. 2(10):65-78 (Oct.) 1968.

Klotz, R., Sherman, J. o., and Egan T.: Preparation of Hyperalimentation Solutions for Seven Pediatric Patients, Amer .. J. Hosp. Ph§.&.. 28:102 (Feb.) 1971.

Elman, R.: Iv1aintenance of Nitrogen Balance by_ the Intravenous Administration of Plasma Proteins and Protein Hydrolysates, Ph.,Y.siol .. Rez.!.. 24:372 19Li.l+~

Dudrick, S. J.: Total Intravenous l~eed:i.ng and Grm·rth in ~ppies, Fed .. Proce. Q_:l~81 1966.

Dudrick, s. J., 1-vilmore, D. i:l., Vars, H .. Me and Rhoao.s, J·. E.: Long Term Total Parenteral Nutrition· v1ith GrovJth, Development and. Positive Nitrogen Balance, Surg,. 64:13L~. (Jul.) 1968. . - -

Wilmore:, D .. vJ. and Dudrick, s .. J .. : Grm·Jth and Development o.f an Infant I?.eceiving all. Nutrients Exclusively by Vein, J .. A.!M.J;. .. 202_:860 (Nar.) 1968.

Dudrick, S. F., l:Jilmore? D .. W., Vars, H. f1. ·and Rhoads, J. E. : Can Intravenous Feeding as a Sole l··Ieans of · Nutrition Support Grm·r:-;h in the Child and Restore Weight IJOSS in an Adult? A...'1. Affirmative Answer, Anno BuJ~o 169: 9?4-8L~ (Jun.) 1969. ---·-- -· ---·

Flack, H. Lo, Gans, J. A., Serlick, s. E. and Dudrick, s. J .. : The Current Status of Parenteral Hyperalimentc-l't;ion~ Amer. ~jf.OSTJ._Pharm. 28:326-35 (I"Iay) 1971.

Du.d.rick, S. J. and Rhoads, J .. E.: New Horizons for Intravenous Feedings, J·.A.Jb_/1. .. 2.12_:939 (Feb.) 1971 ..

107

13.

14.

15.

16.

17.

18.

19.

20.

21.

22.

23.

24.

25 ..

108

Preston, M. J.: Corilparisons of the Various Amino Acid Preparations, C1Jn. Pharmaco1. Ther. 9:61 (Jan.--Feb.) 1968.

Rea, W. J., w. J., andMcC1elland, R. r1.; et al.: Intravenous Hyperosmolar Alimentation, Arch-.-Surg. 100(4):393 (Apr.) 1970.

Vlyrich, W. J., Rea, T.tl. J. and McClelland, R. M.: Rare Complications with Intravenous Hyperosmotic Alimentation, J.A.M.A. 211:1697 (Mar.) 197,0.

\vi1more, D. Vl. and Dudrick, s. J.: Treatment of Acute Renal Failure with Intravenous Essential L-Arnino Acids, Arch. Surg. 99(5):669 (Nov.) 1969.

Elwyn, D. H. and Greenstein, A. J.: A Critique of Parenteral Alimentation itJith Respect to Amino Acid Metabolism, Gastroenterology ~(3):242-57 (Sept.) 1972.

Shils, M. E.: Guidelines for Total Parenteral Nutrition, J.A.M.A. 220(13):1721 (Jun.) 1972.

Stegink, L. D. and B.St.ker, G. L •. : Infusion of Protein Hydrolysates in the .Nevvborn Infant; Plasma amino acid concentrations, J .. Ped. 2.§_(L~):595 (Apr&) 1971..

Dryps, J'ohn S. : Hyperalimen.tation Revie'.1, Drug ];;nformat~_gn Bull.etin, The Grace Hospital, Detroit, Nichigan, 2(1):1-13 (:Bleb.) 1972.

RaviJ.?-, R .. L.: Parenteral _Hyperalimentation, Drug Intell .. ClJ.n. Pharm .. 6:186-89 (Nay) 1972.

Filler, L. J., Jr. , Burness, L. A., Goldbloom, R. B .. , ~t .§.l· : Parenteral :F!'eeding-A Note of Caution, Ped. Lf-9:776-9 (T:iay) 1972.

Klotz, R.: Reducing Bacterial and Fungal Infection Associated with Parenteral Alimentation, Hosn. Phar .. 2=93-4 (Mar.) 1972. ·

Liebert, P. S .. : Central Venous Catheters in ChildrenTheir Placement and Care, Cline Ped~. 10(4) :218 (Apr.) 19?1.

Shadomy, Smith and Shadomy, H .. ~Jean: in Casein Hydrolysate Solution, N. (May) 1972.

Grot·Jths of Candida · Eng. J. IvJed. 286:613

26. \\linters, Ro 1:1~, Santulli, R. V., Heird, H. Ce, et al.: Hype::.·alimentation i'Jithout Sepsis, ~· Eng .. J. 'f,I~cl:--286: 321-22 (Feb.) 1972.

27.

28.

29.

30.

-u--~~~~31.

32.

33.

34.

35.

37.

39.

109

Tachen, M. L. ancl Walther, L. J.: Peripheral Alimentation of the. Small Premature Infant·, J. Ped. 80:895 (1'1ay) 19'?2.

Silvis, s. E. and Paragas, P. D.: Paresthesias, Weakness, Seizures and Hypophosphatemia in Patients Receiving Hyperalimentation, Gastroenterolog;y 62 ( 4): 513 (Apr.) 1972.

Anonymous: Fatal Hyperalimentation Syndrome, Nutr. Rev. ~0~5):121 (May) 1972.

Ghadimi, H., Abaci, F., Kumar, s. and Rathi, M.: Biochemical Aspects of Intravenous Alimentation, Ped. 48(6):955 (Dec~) 1971.

Chan, James c. M., r-1alekzadeh, M .. and Hurley, J.: pH and Titratable Acidity of Amino· Acid f·1ixtures-~used in -~--===-:c_~~~ Hyperalimentation, J.A.l'-1.A. 220(8).:1119-20 (r1ay) 1972. ·

Chan, James C. M., Asch, M,. J., Lin, S., and Hays, D. H.: Hyperalimentation "~dith Amino Acid and Casein Hydrolysate Solutions, J.A.M.A. 220(13):1700-5 (Jun.) 1972.

Karpel, J. T. and Peden, V. H.: Copper Deficiency in Long-Term Parenteral Nutrition, ~. Ped. 80_: 32 (Jan.) 1972.

Danks; M. D .. , Campbell, P. E., Stevens'· B. J., §t aJ.:..: Henke's Kinky Hair Syndrome, Ped. 50(2):188-201 Thug .. ) 1972 ..

Ashcraft, K. \'1. and Leape, L. L .. : Candida Sepsis Complicating Parenteral Feeding, J.A.fvi.-A. 212:454-6 (Apre) 1970.

Travis, S. F., Sugerman, H. J. and Ruberg, R. L.: Red Cell fvJetabolic Alterations Induced by Intravenous Hyperalimentation, N. Eng. J., iVIed., 282(14):763 (Sept .. ) 1971.

Abel, R. l'-1., Abbott, VJ. H. and. J?ischer, J. Eo : Acute Renal ]'ailure :'l'reatment without Dialysis b;:,r Total

_Parenteral Nutrition, Arc~ Sur:& .. 1Q2:513-14· (Oct.) 1971.

Peden, V .. H .. , Hitzeben, C .. J~ .. and Skelton, M. A.: Total Parenteral Nutrition, J .. Fed._ 2.§(1 )_: 180 (Jun.) 1971.

He:i.rd, \'1. c., Nicholson, J. Fq Driscoll, J. M., et al.: Hyperammonemia H.et>ulting from Alimentation Using a Hixture of Synthetic IJ·-Amino Acids: A Preliminary Report, J·- :Ped...!. 81(1) :162-65 (,Julo) 1972.

40.

41.

:42.

43.

44.

45.

1~6.

4'7.

48.

49.

50 ..

51~

52.

110

Caldwell, M.D., Halder, T. J. and othersen, H. B.: Essential Fatty Acid Deficiency in an Infant Receiving Prolonged Parenteral Alimentation, J. Ped .. 81(5):894-98 (Nov.) 1972. -

Ulstrom, R. A. and Bro\vn, D. M.: Hypercalcemia as a Complication of Parenteral Alimentation, J. Fed. 81: 419-20 (Aug.) 1972. · ·--

Allison, J. B.: Nitrogen Balance and the Nutrition Value of Protein, J.A.M.A. 164:283-89 (May) 1957.

Kinney, John M.: Calories-Nitrogen-Disease and Injury Relationships, Drug Intell. Olin. Pharm. 6:261-65 (Jul.) 1972. - -

Rice, C. O., Orr, B., Treloar, A. E., et. al_. : Parenteral Nutrition in Surgery, Arch. ·Sur~. 61:977-91 (Dec.) 1950.

Hyman, c. J., Reiter, J., Rodnan, J. and Drash, A. L.: Parenteral and Oral Alimentation in the Treatment of the Nonspecific Protracted Diarrhea Syndrome of Infancy, Js Fed. 1§112:17-29 (Jan.) 1971.

Mtmro, H,. N.: Basic Conceptf? in the Use of Amino Acids and Protein Hydrolysates for Far-enteral Nutritibn, Drug Intell. Clin. Pl?...~.!.. 6:216-25 (J-un.) 1972. ·

Dudric:h, S., J.: Spontaneous Closure of Traumatic Pancreatoduodenal Fistulas 1:dth Total Intravenous Nutrition, J. Trauma 10:542 (Jul.) 1970.

Hichinar. \'l .. M .. and Lov:, D.: Pa.renteral Nutrition. The Age of.the Catheter, Fed. Olin. N .. Amer .. 12.=373 (I"lay) 1970.

Sinelair, J .. c., Driscoll, J. M., Heird, \'/.c. and Winters, R. i;J. : Supportive Nanagement of the Sick N eo nate: 11arenteral f'Iaintenance, Ped. Cl:i.n .. N. Amer. 17:878-90 (Nov.) 1970. - --

}'iller, R.. M .. , Burness, IJ. A., Go1dbloom, H .. B., &i al. : Total Intravenous Nutrition, .l\ll1§f:.!_ J. S:u_!'g .. 121 :L!·5L4- (Apr.) 1971.

Freeman, J. B. and NacLean, L6 D.: Intravenous Hypera1imentation:A Reviev1, Can. J. Burg. J.L~:180-9L4-0·1ay) 1971.

Benda, G. and Babson, s .. G.: P·eriphera1 Intravenous Alimentation of the Small Premature Infant, J. Ped. 22i22:494 (Sept.) 1971.

53.

111

Dudric~k, s. J. and Ruberg, R. L.: Principles and Practice of Parenteral Nutrition, Gastroenterology §.:J-.(6): 901 (Dec.) 1971.

54. Peden, v. II. and Karpel, J. T.: Total Parenteral Nutrition in Premature Infants, J. P_9d.!-. §.1 (1): 13'7-l.J.l~ (Jul.) 1972.

55.

56.

57.

58.

59 ..

61.

62.

64.

66.

Driscoll, J. !'1., Heird, \1. C., Schullinger, J. N.,, et §.1. : · Total Intravenous Alimentation, J. Ped. 81(12.:145-53

(Jul.) 1972. - -

Helmuth, W. V., Adam, P. A. J. and Sweet, A. Y.: The Effects of Protein Hydrolysate-~1onosaccharide Infusion on IJOv.J-birth-weight Infants, J. Fed. 81(1):129-36 (Jul.) 1972.

Sauve, Sister Frances: The Pharmacist and a Nutritional Intravenous Therapy Program, Amer. J. HosJ2. Phar. 28: 106 (Feb.) 1971. · ·

Kaminski, M. V.: Total Parenteral Nutrition (Hype:calimentation). Prevention and Treatment of Complications, Hyperalimentation Registry, \·.Jalter Reed General Hospital, Washington, D. c., (Nov.) 1972.

Guyton, Art1mr c.: Textbook q:f._M?.gicalJ?]:l;y_siolog;y, 3rd ed.,, \'1., B. Saunders Co .. , Pililadelphia, Pa., 1966, pp. 59-62 ..

Heinemann, H .. 0.: Acid-Base Balance and Intravenous Fluid Therapy' in D-ril;L Is pha£_~~colqgy in, r'1~§-icine., (DiPalma, Joseph R. Jtn.), 3rd Bd., HcGraw-Hill Book Co., New York, 1965' p .. 651+.

Krupp, r'Iarcus A.: Fluid and Electrolyte Disorders, in CJ~~:;s:qE_"t_Di§.e;,gosi£L.£:.:~9: .. J~reatment, Lange fvJ:edical 'Pllblications, IJo~:; Altos, California, 1972, p. 21.

Dudr:i.ck, S. J .. : Rational Intravenoue. Therapy, An:f1r. J·. 1-IOQ.ih_P'Df'J.' .. 28:90 (Feb .. ) 1971.

Welt, I1. G .. and Blythe, H. B.: Cations: Calcium, Magnesium, . Barium, Lithium and. Ammonia, :i.n The __ Pharmacological Be.s~is. £U.h..E?~.SJ?eutics, (Goodman, JJ. s. and Gilman, A. Eds:)i.l-th Ed., The Nacmillan Co., NevJ York, 1970, PPo 805-lL~.

Peleisie~e, B.: Guide to Incompatibilities, ~losJ2. Fhar. 2.: 15-~22 1968 ..

Fo',,Jler, ~l'. tT .. : Irl.Compatibiliti.es of IV Admixtures, Amer!... ;~:!' fl.2.§.I?.!_l.1la.!.!_ _?LJ.:LJ-50 (Aug.) .1967. .

6?.

68.

. 69.

70.

71.

72.

73.

74.

?5.

76.

??.

78.

79.

80 ..

81 ..

112

Welt, , L. , G. and Blythe, W. B.: Anions: Phosphate, Iodide, ]'ly.oride, and other anions, in The Pharmacological Basis. ~rfpeuti£§., (Goodman, L. s. and Gilman, A. eds.) 4th Ed., The 'lacmillan Co., New York, 1970, pp. 819-21.

Swineyard, E. A. and Harvey, s. C.: Gastrointestional Drugs, in Rem~ngtQD's Pharm~ceutical Sciences, (Martin, E. \·!. ed.) 13th Ed.-;l1ack Publishing Co., Easton, Pennsylvania, 1965, p. 881 •

Al-Rashid, R. A. and Spangler, J.: Neonatal Co:pper Deficiency, N. Eng .• J. Hed. 285(15):8LJ·l (Oct.) 1971.

Graham, George G.: Human Copper Deficiency, N. Eng.~ ~ ~85(15):857 (Oct.) 1971. , ,

Hospital Formulary, Letterman General Hospital, The Presidio, San Francisco, California.

Fomon, s. J.: Infant Nutrition, W. B. Saunders Co., Philadelphia, Pennsylvania, 1967, p. 113.

Patai, Saul: The Chemi str;y of the A!l}.ino Group, Interscience Publishers, London, 1968, Po 561.

Guyton, op.· cit., p. 1015.

Jo:O.nson, J .. D$, Albritten, VI. L. and Sunshine, P.: Hyperammonemia Accompanying ParEmteral Nutrition in Nev1born Infants, i[.!-...P~d!. 81~~).:15'+-61 (J\tl.) 19?2.

Guyton, op. cit., p. 115 ..

Edward, Sister ~1ary: pH-An Important Factor in the Com:r;a·ljibility of Additives in IV Therapy, Amer.~ HO.E.J2.!~):~1.ar.. 24:440 (Aug.) 196?.

Greene, Ha:cry r~ .. : Vitamins in Parenteral Nutrition, D:rgg_ Ir-:.:ttll!_ . .Q1.in:_! ___ fl1a~~ 6:355-60 (Oct.) 1972.

Burke, \v. A.: Symposium on Total Parenteral Nutrition, Food Science Committee, Council on Foods and Nutrition . of tho American Nedical Association, Nashville, Tennessee, (Jan.) 1972, p. 181.

Kline~ O. IJ,. and Boehne, J .. vJ .. : Vitamins and Other Nub::ients, in :g_~]i:gg:tQg2 P:g_.:~Efll~eey.t:L~:Q...L Practice..§_ (Osol, Arthur, Chairman Ed .. Board) 14th Ed., Nack Publishing Co., Easton, Pennsylvania, 1970, p. 102L~.

Ame.r-:L.cap. HQ§J?.:it. al_ Fosm:g.lad'.Y_§ervi~ pub. by Amer. Boc. 1iosp. Pharm .. , VJashington, D. c., sections 88:08-24 ..

113

82. T-!J..e_,r:!grck Inde~, (Stecker, Paul G., Ed.) 8th Ed., Merck and Co. Inc., H.ahway, Ne\11. Jersey, 1968, pp. 111.2-5.

83. Abel, R. M., Abbott, vi. M. and Fischer, J. E.: Intravenous Essential L-Amino Acids and Hypertonic Destrose in Patients 1/lith Acute Renal :Failure, ~er. J. Surg. 123: 632-38 (Jun.) 1972.

84. vleisenfeld, S., Podolsky, S., Goldsmith, L., et al.: Adsorption of Insulin to Infusion Bottles and Tubing, piab~tes 17:766-71 (Dec.) 1968 •

. 85. Raf'fanti, E. li'., "Spectrophotomet:r·ic and Microbiological Dete:rmination of the Stability of Sodium A.'Tlpicillin as affected by pH 11 (unpublished r--Iasters 'rhesis, . University

-t------------'·~t'~t-11-@-EMi-f-i-~,~19'/-2 )-;-PP--~-J.G-.~---------------

86. Collin, c. F., et. §.}-.• : Precipitation in the Preparation of Hyperalimentation Solutions Containing Fre1linine - An Electrolyte Study, vlalter Reed Army Institute of Research, \valter Reed Army Medical Center, Washington, D. C.

87. Frye, H. G.: Unpublished Study, through Spencer Laboratory Stockton, California.

88. "The United States Pharmacopeia", 17th rev., Hack Publishing Co., Easton, Po.., 1965, p$ 832.

89. Grove, D.. C. and Randall, VJ. A. : AE?_say ~~th?.£.s _s>:f lmtibj:.2:tJ.c.e., Med.ical Encyclopedia, Inc., NevJ York, N:~, 1955, PP• 192-94o

90. Parker.·, Eugene: Solution Additive Chemica]. Incompatibility· Study, !~r:1er:_. J. Hosp. PheJ& 2L~:L~3L!--39 (Aug.) 1967.

91~ Patel, J. and Phillips, G.: A Guide to Physical Compatibility of Intravenous Drug Aclmb::tures, Arner. J· .. £!.9§.2.!.-~~ g,3..:L!-09-·ll (Aug.) 1966.. -

92 .. Kramer, W .. R.q Inglott, A. s. and Cluxton, R. J .. : Ineompatibilities of Drugs for Intravenous Administration, in l?.~~§~pe,q~ti_y§s in_QJ1--Ei.9B:l- P}]/:l.J:'I]:laq;z:, (Francke, D. E., and· Whitney, J·re, H. A. K. eds.Tlst ed., Drug Intelligenee Publications, Hamilton, Ill., 1972, p:p .. 438-55.

93., Koc:hel, F .. : Incorn.patibilitaten Bei lllischune?;en Int:eavenos(~r Losungen, .Bection Des Pharmaciens Des Hospitaux, JPecle:ca:tion Interna.tionale Pharmaceutique, Sept. 3-lf.~ 1968, pp. 57--70, throttgh Persnec~t.:1·v.0.§._ip.__91.:JJd::..9i~l-!lg\I!!~0.SY, (Francke, D .. E .. and \'/hitney, J·ro, H. Ae K., eds .. ) 1st Ed,., Drug Intelligence Publications, Hamilton, Ill., 19?2, pp. '-~3C3-55 ..

94-.

95.

96.

lllJ.

I1emburg, H.: Compilation of Pharmaceutical Incompatibilities, Hosp. Phar •. 2:19-25 (Aug.) 196? ..

Rao, G. N. R.: Ultraviolet and Visible Spectroscop~, Buttervmrth and Co., Ltd., London, 196?, pp. 190- 2.

Blackburn, Stanley: Amino Acid Determination, Marcel Dekker, Inc., New Y'ork, 1968, p. 182. -

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