Utilizing UPLC for Atline & Online Process Development and

©2011 Waters Corporation | COMPANY CONFIDENTIAL 1

Utilizing UPLC for Atline &

Online Process Development

and Analysis of IPM

Tony Wiklund

Nordic Application Chemist

Craig Dobbs

+1.508.482.2578

[email protected]


Process Analytical Technology

Process Analytical Technology (PAT) is a system for designing, analyzing, and controlling manufacturing processes through timely measurements (during processing) of critical quality and performance attributes of raw and inprocess materials, to ensure final product quality.

— The term analytical in PAT is viewed broadly to include chemical, physical, microbiological, mathematical, and risk analysis conducted in an integrated manner.

— The emphasis in PAT is on the manufacturing process. Quality cannot be tested into products; it should be built-in by design.

— PAT is nothing new, the Chemical Industry has been using it since the 1940‘s.


Inprocess Sample Analysis from Manufacturing


QC Testing Observation in Pharmaceutical Manufacturing

G.K. Raju, MIT PHARMI, Pharmaceutical Manufacturing: New Technology Opportunities, presentation at USFDA 16NOV01


PAT Recommendation for Pharmaceutical Manufacturing

G.K. Raju, MIT PHARMI, Pharmaceutical Manufacturing: New Technology Opportunities, presentation at USFDA 16NOV01


Quality is Paramount, but PAT must also Drive Profitability

Positive impact on business drivers

— Is the Project a ―science experiment‖ or is it financially driven?

o Consistent quality output is an expected baseline

— Well defined ROI (return on investment)

Ma

rgin

/Revenue

Ris

k &

Costs

Asset U

tilizatio

n

Increasing Productivity and Efficiency while Decreasing Costs drive

Profitability


Current PAT implementation

Unit Operation PAT Technology Applied

Benefits in Product Manufacture

Particle size reduction and milling

NIR Spectroscopy Monitor and control API milling process and inprocess agglomerates

Blending and mixing NIR SpectroscopyRaman Spectroscopy

Identify safe-zone and most appropriate mixingtime

Granulation NIR, Raman, XPRD SpectroscopyAcoustic EmissionThermal Effusivity

Justify granulation process parameter ranges with controlled granulation quality

Drying NIR Spectroscopy Identify drying curve and endpoint

Coating NIR SpectroscopyRaman Spectroscopy

Determine coating rate and endpoint

Unit dosing NIR SpectroscopyRaman Spectroscopy

Detect drug distribution uniformity and identify process deviations

Ahmed et al, American Pharmaceutical Review - PAT Initiative in Generic Product Development


Limitations of Current PAT Technologies

Poor Sensitivity

—Dependent on strong compound signal

—Limits of Detection (LOD)

Limited Quantitation capability

—Dependent on dissimilar compound spectra

—Limits of Quantitation (LOQ)

Minimal Dynamic Range

Minimal Resolution

Still requires a ―Reference Standard‖ or ―Gold Standard‖

Technique for comparison and validation

Vibrational spectroscopic data not easily compared/correlated

to data from Discovery, Research, Development, and QC


Product Life CycleAnalytical Support

VibrationalSpectroscopy

DATA and INFORMATICSSpecific to Manufacturing


Product Life CycleAnalytical Support

DATA and INFORMATICS


PATROL UPLC Systems

Embed PATROL Laboratory System Video

Waters Patrol Laboratory System.mov


HPLC for Inprocess AnalysisReference Standard

LC is a major analytical technique in every QC lab

— But it is not a major tool for Online/Atline analysis

What perceptions are impacting the lack of LC adoption on

the production floor?

— Too slow, too slow too slow; not ―real-time‖, it just takes too

long to get an answer

— Long run times delay processes

— Not rugged enough for continuous operation

— Requires considerable maintenance

— Excessive down-time

— Too expensive to operate

— Requires consumables and highly skilled technicians

— Not automated

— Real-time Online/Atline LC Systems don‘t exist


PATROL UPLC Process Analyzer

Key Requirements for an

inprocess analyzer

Real-time results

Ease of Use

Flexible

Rugged & Robust

Serviceability


UPLC SeparationsA New Category of Liquid Chromatography


Increased Speed without Sacrificing Resolution or Selectivity

2.5 µm – 75 mm

F = 500 µL/min

Injection = 2.5 µL

Rs (2,3) = 2.34

5 µm – 150 mm

F = 200 µL/min

Injection = 5.0 µL

Rs (2,3) = 2.28

3.5 µm – 100 mm

F = 300 µL/min

Injection = 3.3 µL

Rs (2,3) = 2.32

1.7 µm – 50 mm

F = 600 µL/min

Injection = 1.7 µL

Rs (2,3) = 2.29

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Original HPLC MethodTOO SLOW for Online Analysis

Transfer to UPLC and optimize

30 minute cycle time


New UPLC Methodreal-TIME LC™ Analysis

Overlay of 10 Replicate Injections

Real-time results

Ease of Use

Flexible

Rugged & Robust

Serviceability


Key Requirements for a UPLC based Process Monitor

Ease of Use

Trade You Seats?


―Typical‖ Analytical LC Workflow

Sample comesinto Lab

Log sampleinto System

4 mins

LabelLC vial

Draw Sample into syringe and aliquot/filter to sample vials

5 mins

Dilute sample andaliquot to LC vial

10 mins

Log sampleinto LC System

& assign method

5 mins

ResultsPlace LC vial in designated

SM position

1 min2 mins

Label sample vials

5 mins

32 minutes 6 potential mistakes


Scan sampleinto System

1 min

―Possible‖ Analytical LC Workflow

Sample comesinto Lab

Draw into syringe and aliquot/filter LC vial

3 mins

Insert LC vial in PSM

2 mins

Results

6 minutes 0 potential mistakes


Design Requirement:Ease of Use – Eli Lilly Implementation Video

Embed Lilly Atline Video


Software – Primary GUI



Real-time results

Ease of Use

Flexible

Rugged & Robust

Serviceability


PATROL UPLC Laboratory AnalyzerPATROL UPLC Process Analyzer

Configured DCS I/O (OPC, 4-20mA)

Thermal management

E-Stop

Buffer, solvent, & wash reservoirs

Waste containers

Detector

Column H/C

Process Sample Manager

Binary Solvent Manager


Online Process Sample ManagerFluidic Components

Process Pump

Process Valve

Sample Pump Inject

Valve

DiluentPump

Priming Valve

Sampling Valve

Courtesy of Bob Tacconi


Process Sample Manager Specifications

Parameter Specification

Sample Identification Online Automatic, Barcode, or Manual

Sample Temperature range 40C - 300C

Minimum Sample Volume 0.5mL (vial residual)

Sample Volume consumed/Cycle <100uL

Injection Volume Fixed loop; 1uL, 2uL, 5uL

Injection Linearity >0.999

Injection Cycle Time <90 seconds, load ahead capability

Sample Carryover <0.01%

Standard Identification Barcode or Manual Entry

Standard Temperature range 40C – 300C


Process Sample Manager Specifications

Parameter Specification

Minimum Standard Volume 0.5mL (vial residual)

Std Volume consumed/Injection <100uL

Dilution Range No dilution to 1:50 programmable

Dilution Precision <1%RSD (caffeine sample)

Dilution Linearity >0.9999 R2 (caffeine sample)

Dilution Time <90secs load ahead capability

Dilution Volume <500uL

Injection wash volume <1.0mL

Injection wash time <0.5mins

Injection, Dilution, Wash Cycle <3.0 minutes

PSM vial positions 32 positions, samples and standards


Atline Dilution CapabilitiesLinearity (n = 6)

R2 = 0.999941/x WeightingExcellent linearity across the entire

dilution range

Aqueous rProtein sample diluted with 6M urea

Undiluted2x Dilution4x Dilution10x Dilution20x Dilution100x Dilution

Dilution %RSD

Undiluted 0.11

2x 0.48

4x 0.70

10x 0.49

20x 0.59

100x 0.66


Online Injections

R2 = 0.99977



Real-time results

Ease of Use

Flexible (H/W & S/W)

Rugged & Robust

Serviceability


Batch Reaction Monitoring by Online UPLC

Map the formation of the

API and the consumption of

the starting material as the

reaction progresses.

The large, linear dynamic

range of the detector allows

for the simultaneous

monitoring of any low level

impurities.

Injection #1

Injection #14

Injection #37

Injection #60


Making a Decision

Monitor API formation to determine endpoint

Process Impurity 1

Process Impurity 2

Process Impurity 3

Process Impurity 4

Monitor appearance of impurities to determine endpoint


Batch Reaction Monitoring by Online UPLC

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Pe

ak A

rea

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Injection #

API Starting Material Process Impurity 1 Process Impurity 2 Process Impurity 3 Process Impurity 4

Map the reaction progress for process understanding and optimization

Quench reaction prior to process impurity 4(CQA) exceeding 0.1%, eliminating additional processing while maximizing yield


Benefits of Batch Reaction Monitoring by Online UPLC

Pro

cess I

mpuri

ty 1

Pro

cess I

mpuri

ty 2

Pro

cess I

mpuri

ty 3

Sta

rtin

g M

ate

rial

API

Pro

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mpuri

ty 4

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0.030

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Injection #25

Quantify low level components in the presence of a high concentration API

Reaction Aliquot

Starting Material

API ProcessImp#1

Process Imp#2

Process Imp#3

ProcessImp#4

1 98.56 1.38

5 89.54 10.43

13 51.03 48.84 0.04 0.06

17 38.49 61.25 0.01 0.10 0.12

25 23.09 75.83 0.05 0.43 0.55 0.04

29 18.27 79.74 0.09 0.81 1.02 0.07

33 14.58 81.96 0.13 1.42 1.77 0.13


Interfacing the PATROL UPLC Laboratory Analyzer to the UNIQSIS FloSyn™ System

A continuous flow reactor can be

directly connected to the PATROL

UPLC Analyzer for online analysis


TODAY

— Collect samples and send to QC lab THEN wait for

analysis results (hours to days)

o Wash solvent analysis

o Swab analysis

— NIR hot-spot analysis

o Technician goes in with probe and measures in the

reactor

o Much faster than laboratory results but still requires

equipment down time

PATROL UPLC

— Wash solvents can be analyzed on-line as they are

removed from the reactor

— Benefits

o ‗Clean until Clean‘ not ‗Clean to Worst Case‘

• Reduced Solvent and Down Time

o Time for Analysis

Online Wash Solvent Analysis to Support Cleaning Validation


Power of UPLC™/UV:One Cleaning Validation Analytical Method

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23

4

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6/7*

8

A pharmaceutical company had 8 individual cleaning

validation methods for 8 different API‘s.

The goal is to develop one analytical method for all 8 API‘s.

With UPLC™, one UV method with LOD‘s from 0.1 to 2 ppm

was successfully developed.

*APIs 6 and 7 are never

manufactured sequentially


Online Wash Solvent Analysis to Support Cleaning Validation

Trial Sample Wash A1

Wash A2

Wash A3

Wash B1

Wash B2

Trial #1 Online 1580 27 — 60 —

Trial #1 Off-Line 1632 31 — 56 —

Trial #2 Online 1647 19 — 40 —

Trial #2 Off-Line 1647 21 — 40 —

Trial #3 Online 1658 29 — 50 —

Trial #3 Off-Line 1678 32 — 51 —

Trial #4 Online 1619 15 — 127 —

Trial #4 Off-Line 1587 17 — 131 —

Swab results were consistent with wash solvents – if swabs were positive for residue, final product was present in the wash solvents

Achieve equivalent results to off-line analysis of wash solvents


Online Wash Solvent Analysis by UPLC Meets the Required Sensitivity Levels

Starting Material R2 = 0.9990Final Product R2 = 0.9996

100ng/mLCompound LOD

(s/n = 3)LOQ

(s/n = 10)

Starting Material 31 ng/mL 102 ng/mL

Final Product 24 ng/mL 80 ng/mL

Results were consistent whether monitored online or off-line for multiple batches

Positive results by swab testing also resulted in positive results for online analysis


Automated Process AnalysisBiopharm Downstream Processing

DOWNSTREAMUPSTREAM


Downstream ProcessingBiopharmaceutical Purity Analysis

COLLECT WASTE

0 12 hours

Main Peak widths are measured in Hours


Original InProcess HPLC Separation

Biopharmaceutical Downstream Analysis

40 minute Cycle Time



0 12 hours


40 minute HPLC analysis

A minimum of 160L

requires reprocessing

Initial process recovery

yield 40%

Final recovery yield after

reprocessing

58%

COLLECT WASTEREPROCESS


PATROL UPLC InProcess SeparationBiopharmaceutical Downstream Analysis

3 minute Cycle Time (+90% Reduction)- No loss of information



0 12 hours


3.5 minute UPLC analysis

No fractions need to be

reworked

Total initial process

recovery yield

of 87% (40%/58%)

6 month ROI

COLLECT WASTE


Process Stream Sampling

For samples requiring physical filtration and/or aseptic

sampling, PATROL can be integrated with multiple

technologies

— Flownamics

— Bayer Technologies

— Sentry IsoLOK

— Nova Biomedical


PATROL System Application Flexibility and Robustness

Real-time results

Ease of Use

Flexible (Applications)

Rugged & Robust

Serviceability


―Hot Swap‖

Prequalified Modules

Metrology Service

Qualification Services

Uptime and Serviceability

Real-time results

Ease of Use

Flexible (Applications)

Rugged & Robust

Serviceability


UPLC for Inprocess Analysis

UPLC enables LC to have “real time” speed

— Routine analysis in <4 minutes

User Friendly

— Doesn‘t require extensive sample handling or prep

— Full automation from sample to report

Large linear dynamic range with excellent selectivity and

sensitivity

— Ability to quantitate major components and low-level impurities

Single technology from Discovery through Manufacturing

— Archived data for Quality by Design efficiency

UPLC performance for use on the Manufacturing Floor


www.waters.com/patrol


Thanks to:

Tanya Sylvain Bob Rob JennJenkins Cormier Tacconi Herbert Burzynski

Documents

Utilizing UPLC for Atline & Online Process Development and