An easy to use fast liquid chromatographic system using a ...s3-us-west-1.amazonaws.com/falconanalytical-net/... · October, 2014 ©2014 Waters Corporation 2 Process Monitoring: Liquid

©2014 Waters Corporation 1

An easy to use fast liquid chromatographic

system using a novel sample manager to

improve work flow for the analysis of

samples close to a manufacturing process.

Charles Phoebe

Principle Applications Chemist

Waters Corporation

Gulf Coast Conference

October, 2014


Process Monitoring:Liquid Chromatography Instrumentation

Primary location for instrumentation “off-line” analytical

laboratories

Viewed as being to “slow” and to “complicated” for real time

release testing

Samples require “preparation” as they are rarely compatible

with liquid chromatographic analysis

Liquid chromatographic systems are not robust enough

However, viewed in many instances as the “gold standard” for

release testing by providing reliable quantitative results based

upon required sensitivity and resolution.


How to Move Liquid Chromatography into Process Monitoring

http://www.analyticjournal.de/glossar_beitraege_einzeln/pa_monitoring.html


First Generation Hybrid Chemistry: 1999 XTerra®

Disruptive innovation allowing for selectivity changes by modifying pH

Si O

SiCH3O

O

O

O

O

SiO

O

O

Neue et. al. American Laboratory 1999 (22) 36-39.


Second Generation Hybrid Idea: Bridged Ethylene Hybrid (BEH)

Si O

SiCH3

CH3

O

O

O

O

O

SiO

O

O

SiO

Si

O

O

O

O

O

Si O

O

O

XTerra®

Si O

Si CH2

CH2

O

O

O

O

O

SiO

O

O

SiO

Si

O

O

O

O

O

Si O

O

O

Bridged Ethylene Hybrid


Waters Particle Technology

Images are on

same scale

(Bar = 10 μm)

5 μm

Analytical Particles

(can fit 12 across hair)

1.7 μm

BEH Particles

60 μm Human Hair

(very fine hair)


How to Realize the Benefit of Small Particles:

Use an LC system that allows us to…..

– Maximize overall system efficiency

o Minimal dispersion flow paths to preserve peak volumes

o Suitably scaled operating parameters (vs. HPLC)

Requires a new generation of LC Systems…..

– Engineered from the ground up in a true holistic approach

o Not enough to modify an HPLC system to handle smaller particles

o Cannot compromise on design to maintain efficiency gains


The Early Days: First System to Work with Small BEH Particles


Change in Technology Landscape:Introduction of ACQUITY Ultra Performance Liquid Chromatography 2004

70’s

1972

M6000

HPLC Pump

1973

µBondapakC18

1978

Sep-Paks

1979

WISP

80’s

1980-84

Major

advances from

integrators to

computers

- Turbochrom

- Maxima

1981

SFC introduced

1983-85API & MS/MS

1985-87

Advances in

laboratory

networking

90’s

1990-92

ESI & PB

1990-91

CE introduced

1993-95Introduction of first relational CDS Millennium

1995HP 1100

1996Alliance

1999XTerra

00’s

2000

ZQ Mass Detector

AutoPurification

2002

Atlantis

Empower

CapLC

2003

Quattro Premier

LCT Premier

Increasing Refinement & Consistency

ACQUITY

UPLC®

Technology

2004


Increased Throughput and Increased Sensitivity

AU

0.00

0.02

0.04

0.06

0.08

0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50

AU

0.000

0.010

0.020

0.00 5.00 10.00 15.00 20.00 25.00 30.0035.00

AU

0.00

0.02

0.04

0.06

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.009.50

AU

0.00

0.05

0.10

0.15

Minutes0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00 1.10

10 µm – 250 mmRs (2,3) = 1.54

5 µm – 150 mmRs (2,3) = 2.69

3.5 µm – 100 mmRs (2,3) = 2.29

1.7 µm – 50 mmRs (2,3) = 2.25

Independent Y-axis

AU

0.00

0.05

0.10

0.15

Minutes0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00

AU

0.00

0.05

0.10

0.15

Minutes0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00

AU

0.00

0.05

0.10

0.15

Minutes0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00

AU

0.00

0.05

0.10

0.15

Minutes0.00 5.00 10.00 15.00 20.00 25.00 30.00

1.10

4.50

9.50

35.00

Note increase in Sensitivity as particle size is decreased

Same Y-axis


Evolution of ACQUITY UPLC Technology

2004 2005 2006 2007 2008 2009 2010 2011 2012

Instrument

Chemistry

Mass Spec Technology

Application Kits

ACQUITY UPLC-Binary-Loop InjectorColumn Oven-Single 65CDetectors-TUV and PDA

ACQUITY UPLC-Sample OrganizerDetectors-Revised TUV and PDA-ELSD

Column Oven-4 column 90C-Heater/Cooler10 – 90C

ACQUITY UPLC-Active CV-Leak sensors-Advanced needle designnanoACQUITY-CapillaryColumn Oven-Single 90CDetectors-Fluorescence

UPLC PATROL-Process AnalyzerUPLC Open Access-open bed autosamplerDetectors-Extended λPDA

ACQUITY UPLC-Local Console Controller

UPLC H-Class-Quaternary -Direct Injector-Met DevH-Class BIO-InertnanoACQUITYHDX-Hydrogen-Deuterium ExchangeUPLC Online SPE

UPLC I-Class-Binary-Loop Injector-Direct InjectorACQUITY UPSFC-SFC / NPUPLC PATROL-Laboratory AnalyzerTRIZAIC-nanofluidicUPLC 2D

ACQUITY UPC2

UPLCAutomated SPE System

Hybrid 1.7 µmBEH C18

Hybrid 1.7 µmBEHShieldRP18BEH C8

BEH Phenyl

Hybrid 1.7 µmBEH HILICBEH 300Å C18

AccQ-Tag Ultra

Silica 1.8 µmHSS T3

Silica 1.8 µmHSS C18

HSS C18 SB

Guard ColumnVanGuard

Hybrid 1.7 µmBEH 300Å C4

OST C18

Hybrid 1.7 µmBEH AmideBEH Glycan

Hybrid 1.7 µmCSH C18

CSH Fluoro-PhenylCSH Phenyl-Hexyl

SFC 1.7 µm2-ethylpyridineFluoro-phenylhybrid

Silica 1.8 µm-HSS Cyano-HSS PFP2.5 µm XP-14 chemistriesSize Exclusion-SEC 200ÅIon Exchange-High Res Q-High Res CM-High Res SP

Size Exclusion-SEC 125Å

Analytical Standards & Reagents

ACQUITY SQDACQUITY TQD3rd party MS control

SYNAPT-Ion Mobility

XEVO TQ MS XEVO QTof

SYNAPT G2

XEVO G2 QTof

XEVO TQ-S

SYNAPT G2-S

XEVO TQD

SQD 2

XEVO G2-S QTof

XEVO G2-S Tof

AccQ-Tag Ultra Amino Acid

Peptide Separations

Oligonucleotide Aflatoxin

Intact Proteins

AccQ-Tag Ultra AAA for H-Class


PATROL: UPLC Technology with a Process Sample Manager

At-line sample

introduction from a vial

On-line sample

introduction from a

process

Sample dilution

Software ease of use

“Green Button”


Atline Analysis From a Vial

Insert a vial into the PSM and click on the Green Icon in Empower to start the Sample Set


Report: Ethanol in Mouthwash


Unique Attributes of the ACQUITY UPLC Process Sample Manager (PSM)

Atline sample introduction from a vial

Online sample introduction from a process



Online sample introduction

Flow Reactor Batch Reactor


Bioreactor and Flownamics FISP Cell-Free Sampling Probe


Automated Process AnalysisBiopharm Downstream Processing

DOWNSTREAMUPSTREAM


Report: Mouthwash Analysis



Atline sample introduction with vials


Sample dilution

– Calibration


Preparation of Calibration Standards


ACQUITY UPLC Process Sample Manager Instrument Method Editor Page: Dilution Factor

Dilution Factor Programmable for each Instrument Method

Available for use in both atline and online modes


Sample Set for Mixed Alcohol Standards: Dilution Factors


Overlaid Standards from PSM Dilution

µR

IU

0.00

3.50

7.00

10.50

14.00

Minutes

4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00

Acetone

Methanol

Ethanol

Isopropanol


Calibration Curves: PSM Dilutions

Are

a

0.0

11000.0

22000.0

33000.0

Amount

0.00 0.45 0.90 1.35 1.80 2.25 2.70 3.15 3.60 4.05

Are

a

0.0

38000.0

76000.0

114000.0

152000.0

Amount

0.00 0.45 0.90 1.35 1.80 2.25 2.70 3.15 3.60 4.05

Methanol: R2 0.99986 with intercept 0.014

Acetone: R2 0.99999 with intercept -0.006

Are

a

0.0

34000.0

68000.0

102000.0

136000.0

Amount

0.00 0.45 0.90 1.35 1.80 2.25 2.70 3.15 3.60 4.05

Ethanol: R2 0.99999 with intercept -0.009

Are

a

0.0

46000.0

92000.0

138000.0

184000.0

Amount

0.00 0.45 0.90 1.35 1.80 2.25 2.70 3.15 3.60 4.05

Isopropanol: R2 0.99999 with intercept -0.005


Report: Denatured Rubbing Ethanol



Atline sample introduction with vials


Sample dilution

– Calibration

– Sample Dilution


Mouthwash with Various PSM Dilution Factors: Sample Concentration Overload

µR

IU

0.00

250.00

500.00

750.00

1000.00

Minutes

2.00 3.00 4.00 5.00 6.00 7.00 8.00

Dilution Factor = 1


Mouthwash: 20 injections at Dilution Factor of 40

µR

IU

0.00

12.00

24.00

36.00

48.00

Minutes

2.00 3.00 4.00 5.00 6.00 7.00 8.00

10 injections from each of two different sample sets

Sorbitol

Ethanol


Results from the Total 20 injection with a Dilution Factor of 40


Ethanol: Gin at Dilution Factor 90


Ethanol: Vodka at Dilution Factor 80


Ethanol: Gasoline Dilution Factor 20



Atline sample introduction from a vial


Sample dilution

PATROL software ease of use “Green Button”

– Barcode directed Empower Sample Set operation


Atline Analysis From a Vial With a Barcode Directed Empower Sample Set


Atline Analysis From a Vial With a Barcode Directed Empower Sample Set

PATROL software allows for the assignment of a specific Sample Set to a specific two digit code on the barcode label


Go to the PATROL Software Page

Click on the “Load Sample” button


Insert the Sample Vial into the Process Sample Manager


Observe the Sample Running


Results


PATROL: UPLC Technology with a Process Sample Manager

At-line sample

introduction from a vial

On-line sample

introduction from a

process

Sample dilution

Software ease of use

“Green Button”


Successful Transfer of Liquid Chromatography into Process Monitoring using UPLC Technology

http://www.analyticjournal.de/glossar_beitraege_einzeln/pa_monitoring.html


Acknowledgments

Aaron Phoebe: Chemical Engineer

Ernie Hillier: Product Manager

Sylvain Cormier: Mechanical Engineer

Scott McLaren: Software Engineer


Documents

An easy to use fast liquid chromatographic system using a ...s3-us-west-1.amazonaws.com/falconanalytical-net/... · October, 2014 ©2014 Waters Corporation 2 Process Monitoring: Liquid