Increased Productivity in Liquid Chromatography Dr. Timothy Cross - - PowerPoint PPT Presentation

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• Dr. Timothy Cross

EMEA Regional Marketing Manager - HPLC

Increased Productivity in Liquid Chromatography

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Benefits of increased productivity Where can we find productivity gains? Hardware enhancements Column technology and chemistries CDS Software Summary

Agenda Outline

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Benefits of Increased Productivity

Higher sample throughput Faster from sample to result Lower maintenance and training requirements Enhanced qualitative and quantitative sample knowledge Improved return on investments Efficient bench space utilization

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Where Can Productivity Be Enhanced?

• System pump pressure
• Thermo Scientific™ Vanquish™ Duo (Dual and Tandem

LC / LC-MS)

• Consistent performance

Hardware

• Smaller column particles and diameters
• Novel chemistries
• Mass spectrometry friendly buffers

Columns and Chemistries

• Thermo Scientific™ Chromeleon™ CDS Software
• Thermo Scientific™ AppsLab Library of Analytical

Applications Software

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620 bar

Thermo ScientificTM UltiMateTM 3000 Basic Automated

• Highly economic &

reliable

Up to 1000 bar 1500 bar

Vanquish Horizon

• High pressure binary

solvent mixing

• Industry-leading precision

and accuracy

• Two thermostatting modes
• Unmatched detection

sensitivity

• Biocompatible
• Integration of multiple

detection technologies Vanquish Flex

• High pressure binary

and low pressure quaternary solvent mixing options

• Two thermostatting

modes

• Biocompatible
• Integration of multiple

detection technologies Thermo ScientificTM UltiMateTM 3000 SD

• Workhorse for standard

HPLC applications Thermo ScientificTM UltiMateTM 3000 RS/BioRS

• Specialty workflow

support

• Binary and Quaternary

UHPLC systems

800-1200 bar

Thermo ScientificTM UltiMateTM 3000 RSLCnano EASY-nLC 1200

• UHPLC systems for

Nano/Cap/Micro range

• Ideal front-end for

proteomics applications

HPLC Systems UHPLC Systems

Routine Analysis Low-Flow Flexibility, More Performance and Method Development Research & HT

Vanquish Duo Workflows

Increasing System Pump Pressure

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Vanquish Horizon Delivers – Boosted MS Performance

Standard Method

Sequence Coverage

100% 100% 100% 100% 100%

Acclaim 120 C18 column, 2.1 x 250 mm, 2 - 45% ACN + 0.1% FA, 60.

5 10 15 20 25 [min] Time (min)

14.64 20.23 10.59 2.73 10.78 14.52 7.99 2.70 6.97 9.43 5.20 1.86 4.45 6.02 3.35 1.13 3.12 4.12 2.43 1.08

30 min gradient 0.4 mL min-1 600 bar 20 min gradient 0.4 mL min-1 600 bar 13 min gradient 0.6 mL min-1 850 bar 8 min gradient 1.0 mL min-1 1150 bar 5 min gradient 1.1 mL min-1 1300 bar

• Vanquish used for high

throughput peptide mapping

• Gradient time reduced from 30

to 5 min with constant 100% sequence coverage

• Thermo Scientific™ Q

Exactive™ HF hybrid quadrupole-Orbitrap mass spectrometer delivers enough data points per chromatographic peak

• Vanquish is able to speed up

peptide mapping significantly

• Vanquish used for high

throughput peptide mapping

• Gradient time reduced from 30

to 5 min with constant 100% sequence coverage

• Thermo Scientific™ Q

Exactive™ HF hybrid quadrupole-Orbitrap mass spectrometer delivers enough data points per chromatographic peak

• Vanquish is able to speed up

peptide mapping significantly

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Three Workflows. Two Flow Paths. One Integrated UHPLC Solution.

Dual LC

• A two channel LC system

with standard footprint

Tandem LC/LC-MS

• System for efficient

detector utilization

Inverse Gradient

• System for advanced

quantification capabilities

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Unique Vanquish Modules

• 2 ternary pumps in one housing
• 1000 bar (15,000 psi) version
• Delivers 2 completely independent

gradients

• 2 injection units in one autosampler
• 1000 and 1500 bar (15,000 or

22,000 psi) version

• Independent flow paths

Vanquish Flex Dual Pump Vanquish Dual Split Samplers

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Thermo Scientific Vanquish Duo for Dual LC

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Analytical Challenge – Thermo Scientific Vanquish Duo for Dual LC

Accelerate time to results Need for efficient characterization

Increased number of samples Process more samples with decreasing number

• f resources

Asked to do more with less

Increased complexity of samples Need for complementary characterization methods

Asked to know more with less

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What Can Be Accomplished With The Thermo Scientific Vanquish Duo for Dual LC? Finished in half the time Efficient characterization

“The instrument offers a fantastic increase in throughput when running two different separation chemistries, enabling us to generate large amounts of characterization data”

• Dr. Jonathan Bones, NIBRT

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Key Benefits – Thermo Scientific Vanquish Duo for Dual LC Easy-to-use as any other Vanquish system

Simple instrument selection Easy method setup with method wizard Automatic method translation

1. 2. 3.

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Key Benefits – Vanquish Duo for Dual LC No additional bench space needed Vanquish Duo for Dual LC system Vanquish system

55 cm 55 cm

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Key Benefits – Vanquish Duo for Dual LC Lower costs per sample Cost of ownership calculator: Throughput increase Cost per sample

Aggregate analysis experimental conditions

Throughput gain Doubled throughput and lower costs per sample

Number of Samples Per Day

96 192

Single Dual

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Thermo Scientific Vanquish Duo for Tandem LC/LC-MS

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Analytical Challenge – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS

Time not acquiring meaningful data

High investments in modern instrumentation Not fully utilizing instrumentation

Asked to maximze the instrument outcome

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Key Benefits – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS Increased Throughput

• Abs. RT Shift

Column left to right [min]

• Rel. RT Shift

Column left to right [%] RT RSD [%]

n=10

Area RSD [%]

n=10

0.023 0.18 0.11 2.47

Vanquish Duo for Tandem

5.0 10.0 15.0 20.0 25.0 30.0 35.0 min 1.6e9 counts 1.6e9

Left column (n=5) Right column (n=5)

counts

LC-MS Peptide Mapping Example

10 20 30 40 50 60 70 80 90 2.1 mm - 50 mm 2.1 mm - 150 mm 2.1 mm - 250 mm THROUGHPUT INCREASE BY (%)

Throughput increase by Tandem LC/LC-MS

5 min Gradient 15 min Gradient 30 min Gradient

“Since installing the Tandem LC-MS system we have doubled our

throughput and increased the precision of our data!”

Richard Rogers, Just Biotherapeutics

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Key Benefits – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS Advanced column wash without sacrificing throughput

50 100 10 20 30 40 50 60 Eluent B [%] Run Time [min] 50 100 10 20 30 40 50 60 Eluent B [%] Run Time [min] 50 100 10 20 30 40 50 60 Eluent B [%] Run Time [min]

Time savings Time savings

Standard gradient profile Tandem mode allows for time savings Extra column washing steps without sacrificing throughput

Additional column wash

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Key Benefits – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS Easy to implement

Automatic instrument recognition and fluidic knowledge Standard instrument method setup Automatic method setup for analytical and reconditioning pump

1. 2. 3.

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Key Benefits – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS Lower costs per sample Cost of ownership calculator: Throughput increase Cost per sample

Long Gradient experimental conditions

Throughput and costs comparison between standard single and Vanquish Duo workflows

Number of Samples Per Day

24 48

Single Dual Tandem

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Thermo Scientific Vanquish Duo for Inverse Gradient

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Analytical Challenge – Thermo Scientific Vanquish Duo for Inverse Gradient Asked to improve detection and quantification capabilities

Need to detect any compound in a sample, even without chromophore Individual standards for each compound required for quantitative measurements

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• Detection of substances

without a chromophore

• Consistent analyte response

independent of chemical structure

• Dynamic range up to four
• rders of magnitude and

highest sensitivity

• Relative quantification using

virtually any standard

Unique Detectors: Charged Aerosol Detection

Comparison of Charged Aerosol Detection to UV and MS

Saving costs for additional detectors or assays

See what others are missing

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Conventional Gradient Elution

Inverse Gradient Compensation for Uniform Response

15% 85% 15%

Eluent dependent analyte response Identical mass o.c.

Analytical Gradient

Inverse Gradient Compensation

15% 85% 15%

Eluent at detector

Analytical Gradient 50% 85% 15% 85% Inverse Gradient

Eluent at detector

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Key Benefits – Thermo Scientific Vanquish Duo for Inverse Gradient

0,0 0,1 0,2 0,3 0,4 10 20 30 40 50 Peak Area (pA*min) Mass on Column (ng) Adenine Tenofovir Emtricitabin Tenofovir Disoproxil 0,1 0,2 0,3 0,4 10 20 30 40 50 Peak Area (pA*min) Mass on Column (ng) Adenine Tenofovir Emtricitabine Tenofovir Disoproxil

Standard free quantification

Without gradient Compensation With Inverse Gradient Compensation

Similar response curves for all analytes after gradient compensation

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Key Benefits – Thermo Scientific Vanquish Duo for Inverse Gradient Easy to implement

Automatic instrument recognition and fluidic knowledge Standard instrument method setup Automatic method setup for compensation gradient

1. 2. 3.

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What is Thermo Scientific™ Accucore™ Vanquish?

• Next generation UHPLC column
• Founded on state-of-the-art Core Enhanced Technology
• 1.5 µm solid core particles
• Designed in synergy with the Vanquish UHPLC to deliver:
• Better Separations: high efficiency enables the separation of very complex mixtures
• More Results: High efficiency at high flow rates
• Easier Interactions: Seamless workflow solution, for simple and easy separations

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• Efficiency = Better Separations

Why 1.5um Provides: Better Separations

0.00 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75 3.00 3.25 3.50 3.75 4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00 min 3 2 1

Accucore Vanquish 1.5 m, 295k N/m Accucore 2.6 m, 196k N/m Fully porous 1.7 m, 176k N/m

40% efficiency gain

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Why Core Enhanced technology Delivers: More Results

40% time saving and Better Separations

Accucore Vanquish C18 1.5 µm 100 x 2.1mm Solid Core C18 2.6 µm 100 x 2.1mm

Average increase in peak capacity of 220% Average increase in resolution of 24%

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Why Core Enhanced technology Delivers: More Results

• 2.0

• 2.0

Accucore Vanquish C18 1.5 µm 100 x 2.1mm C18 1.7 µm 100 x 2.1mm

• Accucore Vanquish v’s 1.7µm competitor solid core column

Average increase in peak capacity

• f 22%

Average increase in resolution of 17%

Why Core Enhanced technology Delivers: More Results

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• Problem: Many areas where manual sequence creation causes errors
• Templates have limited flexibility
• Solution: eWorkflows make it simple for anyone to run powerful analyses

Application Automation – Thermo Scientific™ Chromeleon™ eWorkflows

Select eWorkflow, instrument and launch Set no. of samples and start position Click ‘Finish’

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Proven Efficiency Gains for Lab Productivity

• Significant productivity gains reported by range of customers from

different industries

• Productivity gains have been verified by direct competitor comparison
• Head to head comparison of same tasks (prescribed by customer) in each CDS

Chromeleon CDS – Proven Efficiency Gains

<50%

Windows

<50%

Mouse Clicks

>50%

Faster

Brian Alliston, Sterling Pharma

“Having previously used several other CDS systems, Chromeleon proved to be a very intuitive

• system. It has a ‘familiar feel’ which means it’s easy to learn. The user interface is simple and

clear and everything is where you would expect it to be. I find the system easy to use and training new staff is easy.”

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Thermo Scientific AppsLab Library

Search applications

• Filter based on

instrument, analyte or technique Download eWorkflow

• Hundreds of

eWorkflows Run the eWorkflow in Chromeleon

thermofisher.com/appslab

Find, download, and run new methods in minutes

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Ultra High

Productivity

Liquid Chromatography

Summary

Productivity

• Increased throughput and uptime. Lower costs.

Improved return-on-investment (ROI)

Increased Liquid Chromatography Productivity

• Combination of hardware, chemistry and

software

Next Steps to Increased Productivity

• Discuss productivity solutions at booth 700/704
• See the new Vanquish Duo at booth XXXX

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