UDC 615 Coden: ACPHEE ISSN 1330-0075
PUAI1MAC[UTICA
3/98 PUBLISHED BY - IZDAVAL:
CROATIAN PHARMACEUTICAL SOCIETY
HRVATSKO FARl\IACEUTSKO DRUSTVO
Acta Pharnz. Vol. 48 No.3 pp. 145-219 Zagreb, September 1998
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Acla Pharnz. 48 (1998) 201-209 Original scientific t-UpCl"
The content of trace metals in some herbal teas and their aqueous extracts
RENf\TA SLAVESKA l, The contents of Fe, Mn, Zn, Cu, Ni, Co, Pb and Cd in ILiNKA SPIREVSKA", eleven herbal teas from Macedonia have been investiTRAj(:E STAFILOV', TOMISI_AV RISTOVl gated by atomic absorption spectrometry. The results ob
tained reveal that trace-metal content vary markedly in lFaclilty of Pharmacy different teil-samples and is likely to be higher in teil
University »St. Cyril and MetlIOdills« -leaves than in other tea-samples. It has been shown that
91000 Skopje, Macedonia the content of investigated elements, with some exceptions, followed the pattern: Fe > Mn > Zn ::> Cu > Ni >
2Institlile of Chelliistry Co. Faclilty of Science The determined levels of toxic elements Pb and Cd in all University ),51. Cyril alld MetllOdillS« herbil1-teil samples were regarded ilS basic lcve1s for Pb 91001 Skopje, Macedollia and Cd in plants.
The content of elements ilchieved in the aqueous extracts prepared from herbal teas W,lS also investigated and the results have shown that they vary considerably and do not correspond to that in herb,11 material. In most of the cases, the content of clements in the extracts was in following order Mn > Cu > Fe.
f\l'Cl,j\'L'd Dl'cl'mbef 16, 1997 Keywords: trace metals, determination, atomic absorption ACCl'pIL'd July 17, 1998 spectrometry, herbal teas, aqueous extracts
In plant nutrition studies it is well established that the continuing supply of a certain number of metallic elements is required by plants to maintain their proper growth. Plant species and the representatives within species differ in their use of metallic elements and they respond differently to enhanced or diminished metallic element supply (1,2). The control of health fitness of plants used in nutrition or in medicinal purposes, however, requires systematical monitoring and supervision of some pollutant levels, including toxic elements (3-5).
The information about the distribution of metallic elements in herbal materials is \t'ry useful, since the plants are used as a raw material for pharmaceutical preparation and can also act as a supplement for some elements in nutrition and medicinal treat:~It'nts (6-8).
The aim of this work was to investigate the levels of some essential (Fe, Cu, Mn, Zn, C, J.nd Ni) and toxic elements (Pb and Cd) in some herbal teas. In addition, we analysed :~-2 Fercentage of extraction of some elements in aqueous media as watered-down pre
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201
pJred extrJcts (beverage). The J nJI yses of metJll ic clelTlen ts have been performed by f!Jn1E:' aton,ic ilbsorption spectrometry (AAS) Jnd electrothermJl iltomic absorption spectrometry (ETAAS).
:vrATER1AL X\lD \lETJ-IODS
Herbal material
The sJmples (herbJ] teJs) under investigJtion, consisting of different pl"nt P<1l"ts eJch, <1I"e given in TJble J: three leaf sJmples (No.2, 5 Jnd 7), three flower silmples (No. 3,4, Jnd ]0), two ,1eriJl pJrt sJmples (No. 1, 9), one fruit sJmple (No.6), one mixture of fruits, leJ\'es ,1nd flowers (No.8) Clnd one f,'uit and leave sJmple (No. 11 ). /\11 herbal teCls were produced by Alk"loid, Skopje, MacedoniJ.
TaNI' 1. IIlZJl'stigatl'd IIlcdicilial plallts
S,lmpll' No. _._-"~ ~,_ ..._- ----~-
Phlllt spl'cil's imd plimt pMt --- .._._------- - - -
1 5idcrifis scnrdicn L., ,wriill pilrt
2 Mcntha pipl'rita L., kiln's
3 Matricaria challlol1lilJa L, iJo\\'L'r
4 Tilia sp., ilowcr
5 I\rctostaphylos lIulI-lirsi L., ](>,1\'('S
6 Rosa CI1l1illll L., fruits
7 IhticlI dioica L., k,l\TS
S :V1.1Cedlmi'lll tL'il (Rosa ((7Jlilla I. trui:- "le":::;,; piperita L., !lo'l\'l'S, R,,!'ir:i,; 1",':,,;,1,',1,':': l, . fll)\\l'r)
C) HYIJeri(lllli perl~l!',)tilill L ,ll'ri,ll p,lrt
I I) 5"1111':1(11' ,; L. th1\\'l'r
11 Vllc,'ili ill III 1l;~!rti!l115 L., iruits ilnd ]CilVCS
Sample preparation and {/1wlysis
The Jmount of 30 g of teJ sJmples \"'JS milled to tiny p<1l"ticles (0.5 mm) ilneJ aliquots of 3 g milled sJmple were tJken for eilch JnJlysis. ~~i\'e Jnalyses were pl'rfornwd for e(lch sJmp1e.
For sJmple prep"r"tion (9, 10) two procedures !1iwe been Jpplied.
Procedure 1. - Aliquots u[ dried milll'eJ herb,ll s,lm~lks \\'ere dry-llliner,llized ,1t J temperJture of 400-420 l)C in (I muffle furn(lce with In e\,letl:, controlbble temperilture regiment. The residues obt'lined, (lfter cooling Jt room temperJture, were dissolved in 100 mL of 4% HNO,
Procedure 2. - /-\liquots of dried milled herbJl samples were miner"lized by J mixture of HN03:H20 (I:l/VV) for one to three hours. Further, miner"liz"tion \VJS cJrried
202
out by adding concentrated H 2SO-+ and H20 2 (l:l/V:V). Thereafter, the silmplcs were minerillized at il temperiltUL"e of 400~420 °C in il muffle furnace. The residues obtained were dissolved in 100 mL of 4'>~J HNO~i'
Preparatioll of aqueous extracts. - The aqueous extrilcts of the samp!l' milteriill (1.5-3 g), were prepared according to the given preparation procedure of the manufacturer for each tea-beveri1ge. Namely, the extracts of silmples 1 to 4 ilnd 6 to 10 were obtilined when herbal materiills were soaked in 250 mL boiling deionized w,lter. After ten minutes they were filterL'd. Only, sample 1 "vas filtered after 30 minutes. On the other hilmi, the extraction procedure used for samples 5 ilnd 11 included boiling of the herbal material in deionized water (250 mL) for ten minutes. Afterwilrds, the extrilcts were filtered.
Aliquot of filtri1tes, hereafter designated i1S ilqueous extracts, were eVilpor,lted to dryness, miner,1lized ilt a temperi1ture of 400-420 °C in il muffle furnace. The rest W,lS dissoh'ed in 4'1" HNO:;.
Reagents
Stock solutions (I ~Lg mL-l) of ill! elements investigilted were prepilred using ,111aIytical grade metal chloride or nitrate salts and deionized water.
Instnmlentation
A Perkin-Elnler 703 iltomic absorption spectrophotometer equipped with a Perkin-Elmer HGA-400 gr'1phite furn,lce was used. A deuterium bilckground COHector was ilpplied during the analysis. As light sources hollow cathode lamps were used. PyroJitiCill!y coated graphite tubes were used. Tn order to achieve the optimal conditions, for metallic elements determination bv AAS and ETAAS instrumentill parameters were Vilried. lnstrumental parameters ilre given in TilbJe II.
RESULTS
It WilS shown that the precision of Procedure] (dry mineraliziltion), was more s,1tisfactory for a l! metallic elemen ts eXi1mined than Proced ure 2 (wet-miner,ll iZiltion). Actually, the RSD \'alues for Procedure 1 varied from 3.2 to 6.0% and for Procedure 2 from 1.2 to 7.5'/0. Consequently, in our further work Procedure 1 WilS used.
The iron, mangilnese, copper, zinc, cobalt, nickel, leild and cadmium contents found in eleven tea-samples are summarized in Tilble III, showing that the RSD values varied from 1.2 to 9.9 "1," which are acceptable for the ilmounts investigi1ted.
From the data presented in Table lIT, it is also obvious that the iron level in the Si1mpies vilries markedly (from 46.7 to 513.0 mg kg- l ). The highest amounts of iron ha\'e been found in leaf-silmples. However, the mangi1nese content in the samples investig,lted is lower and varies from 12.2 mg kg-1 to 195.2 mg kg-I. As it can be also seen from Table HI, the most of the samples contain Zn from 17.8 to 34.9 mg kg-I.
Concerning the copper levels in the tea-samples investigated, it has been found that there are no obvious differences between the Cu levels in leaf, flower and fruit samples. Copper content vMies from 4.6 to 16.3 mg kg-I. If we consider the Ni levels of the sam
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R. SLn'eska et nl.: The content of trace JTIet<1ls in some herbill tE'ilS ilnd their aqueous extracts, Acta Pllanll. 48 (l99,s) 201-209
Table II. Instrumental parameters for flame AAS and ETAAS measurements
Element
Parameter for AAS Fe Mn Zn Cu
Wavelength (nm) 248.3 275.4 327.4 213.9
Bilckground corrector Deuterium
Slit (nm) 0.7 0.2 0.7 0.7
Lamp current (mA) 30 20 15 15
Flame Acety lene / air
Element
Parilmeter for ETAAS Ni Co Pb Cd
Wavelength (nm) 323 :2.407 273.3 228.8
Slit (nm) 0.2 0.2 0.7 0.7
Lilmp current (mA) 25 30 10 4
Dry
Temperature (0C) 120
Time (s) 20
Char
Temperature (0C) 900 900 550 300
Time (s) 20
Atomize
Temperature (OC) 2400 2400 2100 2000
Time (s) 5
Cleaning
Temperature (0C) 2650
Time (s) 5
GilS Argon
pies examined they vary from 0.74 (sample 5) to 11.06 mg kg- l (sample 8). The contents of Co in samples range from 0.26 to 0.55 mg kg-1, except in sample 7, in which the value of 1.02 mg kg-1 has been found.
According to our results, the lead content in the samples ranges from 0.1 to 0.3 mg kg-1. Since herbal teas and spices with more than 5 mg kg-1 and 2 mg kg- J Pb, respectively, are considered contaminated with Pb [Official Gazette of SFRJ 59/1983], a remarkably high level of Pb has not been recorded in examined samples.
The content of Fe, Mn, Cu and Pb in aqueous extracts of the examined samples is shown in Table IV. It has been ascertained that Fe could not be extracted over 21.4 '10. Moreover, it has been noticed that lower percentage of Fe extraction (from 1.7 to 7.6%) has been obtained in extracts prepared from the samples with the almost highest content of Fe (such as samples L 3, 5, 6 and 7).
204
R. Sla\'l'ska ct aI.: The content of tf,lCl' mcl~1ls in ~ome herb,1] teas .1nd tJ1eir ,1QUPOlls e\trJcts, Acta Phar11l. 48 (lYY,s) 201-2U9
Table III. Trace metals ill pllllltS slimp/eo
Sam- Content of element (mg/kg-1)
pie Fc Mn Zn Cu Ni Co Pb Cd No. x RSD x RSD x RSD x RSD x RSD x RSD x RSD x RSD
(%) (%) (%) (0'<)) ('1<, ) ('Yo) (':;,) (%)
1 1767 2.0 14.8 4.2 10.8 4.1 10.5 2.1 4.6 4.9 0.3 3.1 0.3 9.9 0.06 9.5
2 397.3 1.8 64.0 3.8 22.1 1.4 13.0 4.4 3.8 2.3 0.5 2.7 0.2 9.8 0.13 5.9
3 345.7 1.6 513 5.2 33.9 3.1 16.3 6.3 3.2 5.7 0.4 6.6 0.2 8.6 0.46 8.3
4 467 66 1094 3.4 17.8 5.2 8.9 54 1.1 7.0 0.4 2.6 0.2 64 0.25 4.6
5 l58.3 2.7 14.8 3.2 33.9 1.8 3.2 4.2 0.7 3.6 0.4 3.5 0.1 9.0 0.39 3.7
6 131.7 2.0 12.2 3.4 20.4 5.0 9.4 2.1 9.3 6.5 0.5 2.2 0.1 3.9 0.39 9.6
7 307.7 3.5 56.8 6.2 20.9 3.2 9.4 2.8 5.2 3.7 1.0 6.0 0.2 2.8 1.78 7.7
8 513.0 1.8 40.0 5.5 9.2 6.5 3.2 12 11.1 1.8 0.6 8.2 0.2 7.9 0.39 6.7
9 91.7 3.4 115.0 6.1 34.9 1.2 8.1 48 1.5 5.2 0.4 4.5 0.2 5.4 0.36 3.0
10 96.7 3.7 35.5 3.8 123.0 3.7 14.6 66 1.9 4.9 0.3 1.9 0.2 2.1 0.13 6.9
11 172.3 2.3 195.2 4.7 28.0 1.4 4.6 3.5 3.6 4.4 0.5 7.4 0.1 8.0 0.51 3.3
x - mean value and RSD - relativc standard deviiltion of five independcnt analyses.
The amount of Mn in extracts has been found to be higher than that of Fe. Thus, Mn might be extracted in good yields (about 30'1'0 in many cases). It should be pointed out that the highest quantities of Mn (almost 84%) have been noticed in sample extract 6. Concerning the content of Cu in the extracts examined, it has been shown that it varies considerably (from 2.7% in the sample 6 to 47.2% in the sample 4). The determination of lei1d content in the sample extracts indici1ted that approximately more than half of the i1queous extracts contains about 50% of Pb present in the herbal m.aterials.
DISCUSSION
Many factors influence the trace metal content in plants, for example the availability and mobility of the trace elements in soil, the pH value of soil, humidity, environmental factors (2), genetic predisposition of a given species to accumulate or to exclude metals, period/season of sampling, plant part, influence of herbicide treatment (3) and others.
Iron and manganese are two well-known essential elements and their biological roles have been extensively studied. A continuing supply of iron is required by the plants to maintain proper growth. Plant species and varieties within species differ in their use of iron (11). Thus, when iron values are 50 mg kg-lor less in mature leaves, deticiency is likely to occur. The sufficiency range seems to be from 50-250 mg kg-I Fe (12). Since the Fe concentration in young plants can be extremely high, reaching levels higher than 300-400 mg kg-1 (12), our results suggest that the samples 2 and 7 containing 397.3 and 307.7 mg kg- 1 Fe, respectively consisted of young leaves. A health risk due to the high iron content is unlikely because Fe has only low toxicity. The manganese deficiency normally occurs when the plant tissue concentration is less than 20 mg kg-1 in the dry
205
-- - -------- -- --- --
--------
N o 0'
Tablc IV COlltClltS of Fc, Mil, ClI alld Pb ill aqllcolls extracts like IJerl'rll teas
---~ ._----~------ _._~._._- ------- -.-------- --_.-- ---_._.~._-------- - - -.-------
fe Mn ell Pb ------------- - --------_. - - ------- -"----- "_._---
Sample v y y No. (mi L-1) mg kg-1 Ext. ('X,) (mg L-1) mg kg-1 Ext. (%) (mg L1) mg kg I Lx!. no) (mg L-1) mg kg 1 Ext. r>,,) I .----_._- -.-------------- - ---_.__.--------- -- .------ - - ---------~--.-_._------ I
1 0.31 7.75 4.4 0.16 4.00 27.0 1.10 2.75 26.2 0.012 O.lS 5R.8
2 1.25 4167 10.5 0.61 20.33 31.7 142 4.73 ~(l.~ 0006 O.OOR 41.2
3 0.27 8.00 2.3 032 10.67 20.8 0.29 0'11) h.O 00U6 0006 29.6
4 0.19 6.33 136 0.61 2033 1R.6 1.26 42 rc; U.006 (ll 51.2
5 0.48 1200 7.6 020 5.00 33.8 0.12 0.3 '1.·l < 0001 11.001 < 1.0
6 0.54 464 35 OS] 10.20 83.6 0.13 O.2h .J\ < 0.001 0.001 < 1.0
7 0.27 532 17 U.2S 9.33 1b.4 02S Il.'il III() 0.1 0.13 55.6
.~() [18 0.81 90.1S 17.6 1.2U 24.00 60.0 059 liS 0008 0.08 46.3
9 0.43 14.30 156 0.94 31.33 27.3 0.83 2.77 ~·I .:) (l.00() 0.1 51.2
10 031 20.67 21.4 o15 1l1.00 28.2 0.27 l.SO I.' I, ootn 01 512
11 1.11 27.74 16.1 0.70 17.50 9.0 0.25 O.hl 1\(, o.()Ol 0.002 18.3
Ext ('r,,) - percentage "f extraction on dry mass b,1,is
?= '£: '" r;, r ~
c.
..., :0G
r.
:; G :;
~
'f
'f C :; G
! z ;;:7 ~
::,
.;' ~
:; 'f
;;
~
j.. c.
-0,
..... cP
f
S'J
'J,
:~ C
I{. SLl\'eskc1 ('f aI.: The content nf tf.lCl' mL't<1b in ~oml' ]1l'rb,11 tea ... Jnd thelf ~lqlll'ollS l'>..tr<tds, Acta fJ11l1rJl! . .I8 (1l)4H) 2Dl-2(ll)
matter (12). Mn values obtained are lower than the toxic level (more than 500 mg kg-I Mn) of many plant cells (12).
In general terms, zinc is essential to the multiple process of plant (13). Zinc deficiency occurs in many plants when leaf concentration is less then 20 mg kg-1. The normal concentration is in the range of 25 to 50 mg kg-1 Zn. Toxicity occurs 'when the leaf Zn level exceeds 400 mg kg-1 (12). The Zn levels found in samples plant leaf showed no value above the toxic level. Namely, the concentration varied from 9.3 to 123.0 mg kg-I.
Copper was demonstrl1ted to be l1n essential elernent for plant growth and it has been shown to act as an important filctor in severol biochemical processes (14). The normal range for Cu concentration of pll1nt tissue is l1bout 5 to 20 mg kg- 1. When the Cu concentrl1tion in pll1nts is lower than 4 mg kg-1, deficiency is likely to occur. However, when Cu levels are higher than 20 mg kg-1 in ml1ture leaves, toxicity may appear (11). It is obvious that levels of Cu in investigated teas are within the normal concentration levels except for 11 few samples.
The levels of the elements investigated (Fe, Mn, Cu l1nd Zn), recognized as essential elements for plant metabolism, arc in almost the same range of milgnitude as reported by other l1uthors (7, 15-17).
The nickel concentriltion in plonts growing in soils other than that with excessive nickel levels are usually lower thl1n 10 mg kg- 1 (18). The essentil1llity of nickel for the proper grO\\'th of plants hl1s not been yet proven. Among sl1mpJes investigl1ted only SillTlple 8 possesses excessive Ni level (11.06 rng kg-1).
TriKes of cobl1lt are rebtively non-toxic to animals and human beings. As l1cknowledged, cobl1lt is biologically active and directly associated with the hel1lth of some plants, animl1ls and hum.an beings (11). A deficiency of cobalt is of a greilter concern than potentially toxic concentration in plants. No positive correlation has been shovvn between total soil cobalt l1nd the l1mount of Co in pll1nts. The cobalt content of \'l1fious plant species, when grown under the Sl1me conditions varies markedly (11). However, our results show that there are no great differences between the content of Co in the samples (Table III).
It is \\'el1 known that lead l1nd cl1drnium have been estoblished as verv toxic elements, environmental pollutilnts v:ith no physiologiccll function. Le<1d is 'Iv<1ibblc to plants frorn soil l1nd l1erosol sources. The l1ctu111 uptl1ke ,md ilssoci'ltion of lead with pbnts l1re "ffected bv <1lmost <111 environmental factors. 1t lias been observed thl1t the lead content of pl<1nts generally reflects the extent of biologically aV<1ibble le<ld (19). Plants growing neilr highways ilre usuilily exposed to more le,ld than 1110St other 10cl1tions (13) and it is almost ccrt,lin that such pl'lnts possess elevated lead levels. The lead values obtained in the samples im·estigated Me regarded ,IS the basic levels for le'ld in medicinal plants (T,lble III).
Geochemically, cadmium is a rare element. Bec<1use of the low soil Cd leve], it is not considered lUI essential element for plant growth. The studies on the Cd content of plant segments, resulting from the data, indicate potenti111 health hazards of Cd (21). The absorption of cadmium by plants has not been extensively studied. Among all physiologiIral segments of the plant, seeds are usually the lowest in the content of Cd, and in the Ile'lf it is higher thelll in fruits and roots (22, 23). As it can be seen from Table HI, the Iplants ilwestigated are not contaminated \vith Cd.
R, Sl'l\'psh.a l'I £11,: The content of tr<lce I11L't,1b in some herb.1! tl'<l5 <1nd their ,1qUl'UlIS c;...tr<lcts, Acta [JJWI'IIl. 48 (1LJ(J:--l) 201-'2U9
It is apparent from the results obtained that the percentage of extraction of Fe, Cu, Mn and Zn vi1ries considerably in different samples (Table IV). Besides, the vzllues of Fe, Cu, Mn and Zn in crude plant material do not correspond to the obtained levels in plilnt extracts. Thus, the amount of Fe achieved in extracts ranged from 1.7'iS to 21.3'!i, of the content in crude herbal materiill. Most significant differences in the percentage of extraction have been noticed for t'vin and Cu ranging from 9.0')';, to 83.6% and from 2.8')!" to 47.2%" respectively. Almost :JO% of crude herbal content of lead is extracted. It is necessary to point out that different studies have shown that ascorbic or citric acid em improve the extraction of Fe, Mn, and Zn from leaves of Achillea 11lillefoliulIl (24) and Sal"uia officinalis (25).
CONCLUSION
Since the distribution and accumulation of metallic elements in plants is a very complex process, it is hardly possible to correlate the content of metallic elements with all the above mentioned fi1ctors that influence their concentration in plants (the availability and mobility of the trace clements in soil, pH villue of soil, humidity, genetic predisposition, environmental factors Llnd others) with no additional information about the samples investigated.
Our investigation, hO\-vever, shows that the content of elements in the herbal samples is likely to be higher in leJf sanlples than in other plant parts, in accordance with the intensive metabolic activity of leaves (chlorophyll metabolism cmd photosynthesis). Summarizing the contents of the elements investigated, vvith some exceptions, it follows the pattern: Fe > ;yin> Zn ~ CLl > Ni > Co. It has been also shown that tea mixtures exhibit almost the highest level of investigated elements.
The content of pollutants Pb Llnd Cd in the herbal samples investigated does not show values above the allowed limits (up to 2 mg kg-1).
Concerning the percentage of extrzlCtion of Fe, Cu, Mn ilnd Zn, it can be ilssumed that it varies markedly. The amounts obtained in the extracts do not correspond well "vith the levels determined in the crude herbill material. The amounts of Fe, Cu, Mn ,1l1d Zn achieved in most cases of aqueous extrilcts obeyed follovving order: Mn > Cu > Fe. Thus, although iron was the most abundant element in the crude herbal material investigated, when extracted it was in the lowest percentilge. The leild was extrilcted to illmost 50% of its content in herbill mLlterial. When trilce elements in herbal materiJl Jre concerned, additional experiments must be cJrried out to estilblish the optil11Lll extraction procedure for eJch element of interest.
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SAZETAK
Metali u tragovima u nekim cajevima i vodcnim ekstraktima biljaka
Metodom atonLske apsorpcijske spektromctrije odredivan je sadrzaj Fe, Mn, Zn, Cll, Ni, Co, Pb i Cd II jedanaest C<ljCV<l pripravljenih od biljakCl iz Makedonije. Rezultati ukazuju da kolicina n<lvedenih metZllZl znacZljno vMira zavisno 0 uzorku biljke, dZl je veea II
listovima nego II drugim dijelovima biJjaka te, llZ neke iznimke, prati redoslijed Fe > ]\1n > Zn ::> Cll > Ni > Co. KoJicina II vodcnim ekstraktima takoder varirZl i u veCini slllcajeva slijedi niz Mn > Cu > Fe. Kolii'ine toksicnih elemenata Pb i Cd ne prelZtze osnovnu razinu.
KIJucne rijei'i: tragovi metaL1, odredivilnje, atulllska apsorpcijska spektrometrij,l, caj, vodeni ekstrakt
Faculty of Plzarlllacy IIlld Faclilty uf Sciellce Ulliversity »St Cyril alld MetlzodiIIS<', Skopje, Macedollia
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