Analytical Instrument Qualification According to …...Analytical Instrument Qualification According to USP : Reqqpuirements and Examples for the Agilent 1290 Infinity

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!


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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


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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


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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


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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


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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


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Thank you for your attention!Thank you for your attention!

D h Q ti ?Do you have Questions?


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Analytical Instrument Qualification According to …...Analytical Instrument Qualification According to USP : Reqqpuirements and Examples for the Agilent 1290 Infinity