Analytical Instrument Qualification According to USP <1058>:
Requirements and Examples for the q pAgilent 1290 Infinity LC System
1) USP <1058>: Scope, approach, requirements2) Qualification Examples for LC 12903) Selected performance tests3) Selected performance tests
Dr. Edgar NaegeleApplication ScientistAgilent Technologies, Germany
February 22, 2010Application title
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Design Qualification S l t d HPLC ifi ti f d i lifi tiSelected HPLC specifications for design qualification.
Selected specifications of theA il t 1290 i fi it t lAgilent 1290 infinity autosampler
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Design Qualification S l t d it i f d tSelected criteria for vendor assessment.
XX
XX
XXXXX
All requirements of this custom vendor assessment template
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are fulfilled by Agilent Technologies!
Installation Qualification1) Template and examples to document1) Template and examples to documentcompleteness of shipment for IQ. 3) Equipment documentation for IQ.
yesyesyes
2) Verification of construction requirements for IQ.
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Installation QualificationTesting for Installation Qualification
Verification of correct system installation for IQ.
For this purpose Agilent p p guses a certified standard!
February 22, 2010Application title
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Operation qualification - an example procedure
Pump Injector Oven DetectorComputerControl&
HPLC - System
Signal
Pump Injector Oven Detector Control&Evaluation
Flow rate Inj Vol Temperature Signaland/or noise
Flow rate precision
Inj. Vol.accuracy
Temperature accuracy
Specifications of the Agilent 1290 infinity LC systemp g y y
Flow rate precision
Inj. Vol.accuracy
Temperature accuracy
Signaland/or noise
0 07% RSD or < 0 25 RSD +/ 0 8 °C < +/ 3 x 10-6 AU0.07% RSD or 0.005 SD at 0.2 – 5.0 mL/min
< 0.25 RSD from 5 – 20 µL< 0.50 RSD from 2 – 5 µL < 0.70 RSD
+/- 0.8 CPrecision+/- 0.05°C
< +/- 3 x 10 6 AU at 230/4 nm, slit width 4nm, TC 2 s ASTM
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from 1 – 2 µL
Operation qualification - an example procedureComputer Control & Evaluation
Generated Automatically
Header
Signed and datedSigned and dated
Summary
Test/Limit/Result/Status
C fi tiConfiguration
Module/FW/Serial No. & Software
Verification Log Book
Test Reports
Sequence/Log
Method
Chromatograms/Spectra/Reports
Results/Sample Summary
February 22, 2010Application title
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Gradient test depending on the volume of the mixers (blue-35µl, red-135µl, green-300µl)( µ , µ , g µ )
mAU
600
Time % A % B0 00 100 00 0 00
Gradient (stepwise):
400
500
600 0.00 100.00 0.003.00 100.00 0.003.01 90.00 10.006.00 90.00 10.006.01 52.00 48.009.00 52.00 48.009.01 48.00 52.00
200
300
9.01 48.00 52.0012.00 48.00 52.0012.01 10.00 90.0015.00 10.00 90.0015.01 0.00 100.0018.00 0.00 100.0018.01 48.00 52.00
min0 5 10 15 20 25
0
100
8 0 8 00 5 0021.00 48.00 52.0021.01 52.00 48.0024.00 52.00 48.0024.01 100.00 0.0027.00 100.00 0.00
Influence of the mixer
Gradient Testing:Mobile Phase A: Water Mobile Phase B: Water + 0,5% Aceton (v/v)
1290 Infinity Binary Pump
„Next Generation“ Technology
1200 bar @ 2 0 mL/min
Jet WeaverMixer
Solvent selection valve
Degasser
High resolutiond i 1200 bar @ 2.0 mL/min
800 bar @ 5.0 mL/min
Minimum dwell volume: 10 µl / 45 µl
Minimum „Mixing Noise“
Multi layerheat exchanger
pump drives
Minimum „Mixing Noise
Maximum RT repeatability
Automatic „Purge“
Automatic correction of von micro leaksSilicon Carbide
Channel A
Channel B
Purge valvepiston
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1290 Infinity binary PumpMinimum dead volume and mixing noiseg
1200 SeriesSolvent A
Passive Damper0 - 300 µl
Passive Mixer400 µl
1200 SeriesSolvent B Low Noise
~ 800 µl dead volume
1290 InifinitySolvent A
1290 InifinitySolvent B
10 µl / 45 µl DV:80x / 20x kleiner
Niedriges „Noise“< 50%
=10 µl / 45 µl DV:
20x smallerLower „Noise“
< 50%
Active Damping: 10 µl
FirmwarePump +
HardwarePump
Jet Weaver Mixer: 35 µl
+ Agilent microfluidic, Multi-layer Technologie
< 50% < 50% u p p y g
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Example: Performance QualificationD t ti f PQ t tDocumentation of PQ tests.
ChemStation Report with detailed statistical information
AmountS.D. : 0.035RSD: 0.13
February 22, 2010Application title
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Performance qualification – an exampleAUmAU
500
600
6850.
451 0.55
1 Peak RSD % RT (n=10)
RSD % Area (n=10)
1 0.01600 0.4012 0 00769 0 393
400
500
80.
7610.
617
0.71
30. 2 0.00769 0.393
3 0.00517 0.4294 0.00575 0.4705 0 00577 0 398
200
300
0.88
90.81 5 0.00577 0.398
6 0.00538 0.4637 0.00585 0.437
0
1008 0.00571 0.5189 0.00797 0.555
Mean 0.00725 0.450
min0 0.2 0.4 0.6 0.8 1
Sample: Check-out sample with 9 phenones
Column: 2.1 x 50 mm, 1.8 µmGradient: 5 to 95 % B in 0.5 minFlow rate: 1.2 ml/min
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Sample: Check out sample with 9 phenonesInjection volume: 1µL
Analytical Instrument Qualification According to USP <1058>:
Requirements and Examples for the q pAgilent 1290 Infinity LC System
1) USP <1058>: Scope approach requirements1) USP <1058>: Scope, approach, requirements2) Qualification Examples for LC 12903) Selected performance tests and performance data
February 22, 2010Application title
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Application depended Performance Data and System Suitability - Experimental conditions for the Agilent 1290 infinity LC system
Configuration of the 1290-SystemG4220A 1290 Infinity Binary pump with
integrated vacuum degasser and different solvent mixers
Reference samples A liquid formulation of metoclopramide hydrochloride (1 mg/mL) comprisingdifferent solvent mixers
G4226A 1290 Infinity AutosamplerG1316C 1290 Infinity Column Compartment
G4212A 1290 Infinity Diode Array Detector
Software: Chemstation B.04.02
hydrochloride (1 mg/mL) comprising 0.01% concentration of impurities was used for all tests.
Setup for testing1. Determination of pump performance depending on dwell volumes by gradient tests2. Establishment of a chromatographic separation to achieve data for long time evaluations3. Similar peak pattern and resolution according to selected column with respect to particle size and column dimension and adapted to gradient shape and flowcolumn dimension and adapted to gradient shape and flow4. Precision of areas must be < 2 % RSD. 5. Precision of retention times must be < 0.5 % RSD.6. Linearity should be given at least with R2>0.999
Sample setup for testingSample Purpose InjectionsBlank solution Verify baseline stability and identify
artifacts3
Suitability sample Verify precision of areas andretention times for reference solution
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Calibration Verify linearity 3 for each levelHi h t t ti V if C 3 f h l
g
February 22, 2010Confidentiality Label
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Highest concentrationand Blank solution
Verify Carry over 3 of each sample
Separation of Metoclopramide and its impurities on different columns.
A B
C D
A) Agilent TC C18, B) Eclipse Plus C18, C) Eclipse Plus C18, D) Waters BEH C18,A) Agilent TC C18,250*4.6mm, 5µm
B) Eclipse Plus C18,150*2.1mm, 3.5µm
C) Eclipse Plus C18,150*2,1mm, 1.8µm
D) Waters BEH C18,150*2.1mm, 1.7µm
Flow rate 1.058ml/min 1.058ml/min 0.221ml/min 0.221ml/minGradient 0-25min 5-57% B 0-15min 5-57% B 0-15min 5-57% B 0-15min 5-57% BTemperature 37°C 37°C 37°C 37°CInjectionvolume
8µl 2µl 1µl 1µl
Detection DAD, Signal 275/4, Reference 400/60, standard cellData rate 2 Hz 2 Hz 40 Hz 40 Hz
Mobile phase A : 0.25% w/w Ammonium acetate in water Mobile phase B : Acetonitrile
Data rate 2 Hz 2 Hz 40 Hz 40 HzMaximumpressure
138 bar 145 bar 310 bar 435 bar
Determination of the precision of areas and retention times – Impurities in Metoclopramidep
Retention times Areas LinearityMean RSD Mean RSD R2
Impurity 1 6.927 0.030 3.765 1.36 0.9994Impurity 2 7.386 0.025 2.170 1.49 0.9997
Determination of the precision of areas and retention times
p yMetoclopramid 8.170 0.039 10190.182 0.58 0.9998Impurity 3 8.714 0.035 1.707 1.48 0.9993Impurity 4 9.395 0.013 1.496 1.58 0.9993Impurity 5 10.018 0.039 1.023 1.55 0.9990Impurity 6 10.551 0.039 1.971 1.46 0.9993
February 22, 2010Application title
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Agilent 1290 Infinity AutosamplerFlow-through designg g
Advantage of the flow-through designN l l•No sample loss
•Flexible injection volume with high precision from 0.1 to 20 µL•No need to exchange the sample loopCompromise of the flow-through design•„higher“ dead volume of 85 µL•For the 1290 Agilent infinity LCFor the 1290 Agilent infinity LC system 125 µL dead volume if loop is in main path
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Detection of carry over. A) Injection of sample with highest concentration of metoclopramide. B) Injection of g p ) ja blank solvent sample. C) Carry-over specification.
A Metoclopramide mAU
DAD1 A, Sig=257,4 Ref=360,100 (F:\CHLORHEX100\WATER1200000014.D) DAD1 A, Sig=257,4 Ref=360,100 (F:\CHLORHEX100\CHLOREX30020013.D)A Metoclopramide mAU
1
1.25
1.5
Chlorhexidine
B0
0.25
0.5
0.75
min0.25 0.5 0.75 1 1.25 1.5 1.75 2 2.25-0.5
-0.25 0.0028% carry over
Column: 2.1x50mm Eclipse Plus C18, 1.8µmSample: Chlorhexidine (1200ng/µl) dissolved in Water+0.1%TFASample: Chlorhexidine (1200ng/µl) dissolved in Water 0.1%TFAMobile Phases: water+0.05%TFA , Acetonitrile+0.045%TFAIsocratic: 64 % A + 33 % B. Flow rate: 0,5ml/min, Stop time: 2.5minInjection volume: 1µl, external needle wash, water + 0.1%TFA, 10secColumn temperature: 50°CDAD: 257/4nm, Ref 360/100nm, 20Hz
Speed optimized separation of Metoclopramide and its impurities p
Zorbax Eclipse Plus C18 150*2 1mm 1 8µmZorbax Eclipse Plus C18, 150 2.1mm, 1.8µmFlow rate : 0.44ml/minGradient : 10 minutes
1290 Infinity Diode Array Detector Optofluidic Waveguides
Non-coated fiber (fused silica)
Advantage
High light transmission by „Total-Internal Reflection“ (TIR) Principle
AdvantageHigh Sensitivity at Lowest Cell VolumeNo RI (Refractive Index) and Thermal Effects (Solvent Temperature)Data Rate up to160 Hz (Multi Wave Length + Spectra)One Flow Cell for all Applications
Cartridge Design for easy Handling
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g g y g
ConclusionThe new Agilent Infinity LC is designed to provide highest speed, resolution and sensitivity.
A new power range allows you to operate with any particle type, any column dimensions, or any mobile and stationary phase.
The Agilent 1290 Infinity HPLC system is can be qualified according to the USP <1058> chapter and will meet highest requirements for every LC application.
A selection of tests that can be established to evaluate a LC system were shown: For instance gradient testing, the determination of precision of areas and retention and the determination f li itof linearity.
All results show explicitly the applicability of the 1290 Infinity LC system for quality control testing as well as in the FDA regulated environment.
Agilent Services qualifies your system according to USP1058!
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References/Further reading• United States Pharmacopiea Chapter <1058>• United States Pharmacopiea, Chapter <1058>,
Analytical Instrument Qualification, USA, 2008
• Analytical Instrument Qualification and System Validation January 1 2009 Agilent PublicationValidation, January 1, 2009, Agilent Publication Number 5990-3288EN
• Increasing productivity in the analysis of i iti i t l id h d hl idimpurities in metoclopramide hydrochloride formulations using the Agilent 1290 Infinity LC system, Agilent Technologies Pub.-Number: 5990-3981EN
• Analytical instrument qualification and system validation according to USP Chapter <1058> for the Agilent 1290 Infinity LC system, Agilentfor the Agilent 1290 Infinity LC system, Agilent Technologies Pub.-Number: 5990-4781EN
February 22, 2010Application title
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