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

Increased Productivity in Liquid Chromatography Dr. Timothy Cross EMEA Regional Marketing Manager - HPLC The world leader in serving science

Agenda Outline Benefits of increased productivity Where can we find productivity gains? Hardware enhancements Column technology and chemistries CDS Software Summary 2

Benefits of Increased Productivity Faster from sample to result Higher sample throughput Efficient bench space utilization Improved return on investments Lower maintenance and training requirements Enhanced qualitative and quantitative sample knowledge 3

Where Can Productivity Be Enhanced? Hardware • System pump pressure • Thermo Scientific™ Vanquish™ Duo (Dual and Tandem LC / LC-MS) • Consistent performance Columns and Chemistries • Smaller column particles and diameters • Novel chemistries • Mass spectrometry friendly buffers Software • Thermo Scientific™ Chromeleon™ CDS Software • Thermo Scientific™ AppsLab Library of Analytical Applications 4

Increasing System Pump Pressure HPLC Systems UHPLC Systems Flexibility, More Performance and Method Routine Analysis Low-Flow Research & HT Development Vanquish Duo Workflows Vanquish Horizon •High pressure binary Thermo Scientific TM solvent mixing UltiMate TM 3000 •Industry-leading precision Vanquish Flex RSLCnano and accuracy •High pressure binary Thermo Scientific TM •Two thermostatting modes EASY-nLC 1200 and low pressure UltiMate TM 3000 •Unmatched detection quaternary solvent •UHPLC systems for RS/BioRS sensitivity mixing options Nano/Cap/Micro range Thermo Scientific TM •Specialty workflow •Biocompatible •Two thermostatting UltiMate TM 3000 SD •Ideal front-end for Thermo Scientific TM support modes •Integration of multiple proteomics applications UltiMate TM 3000 Basic •Workhorse for standard •Binary and Quaternary detection technologies •Biocompatible Automated HPLC applications UHPLC systems •Integration of multiple •Highly economic & detection technologies reliable 620 bar 800-1200 bar Up to 1000 bar 1500 bar 5

Vanquish Horizon Delivers – Boosted MS Performance • Vanquish used for high • Vanquish used for high Sequence Coverage Standard Method 14.64 throughput peptide mapping throughput peptide mapping 30 min gradient 20.23 0.4 mL min -1 10.59 2.73 100% 600 bar • Gradient time reduced from 30 • Gradient time reduced from 30 10.78 to 5 min with constant 100% to 5 min with constant 100% 14.52 20 min gradient sequence coverage sequence coverage 7.99 0.4 mL min -1 2.70 100% 600 bar 6.97 9.43 13 min gradient 5.20 • Thermo Scientific™ Q • Thermo Scientific™ Q 0.6 mL min -1 100% 1.86 850 bar Exactive™ HF hybrid Exactive™ HF hybrid quadrupole-Orbitrap mass quadrupole-Orbitrap mass 4.45 6.02 8 min gradient 3.35 spectrometer delivers enough spectrometer delivers enough 100% 1.13 1.0 mL min -1 data points per data points per 1150 bar chromatographic peak chromatographic peak 3.12 4.12 100% 5 min gradient 2.43 1.1 mL min -1 1.08 1300 bar • Vanquish is able to speed up • Vanquish is able to speed up 5 10 15 20 25 [min] peptide mapping significantly peptide mapping significantly Time (min) Acclaim 120 C18 column , 2.1 x 250 mm, 2 - 45% ACN + 0.1% FA, 60 � . 6

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 7

Unique Vanquish Modules Vanquish Flex Vanquish Dual Pump Dual Split Samplers   2 ternary pumps in one housing 2 injection units in one autosampler   1000 bar (15,000 psi) version 1000 and 1500 bar (15,000 or  Delivers 2 completely independent 22,000 psi) version  gradients Independent flow paths 8

Thermo Scientific Vanquish Duo for Dual LC 9

Analytical Challenge – Thermo Scientific Vanquish Duo for Dual LC Need for efficient characterization Accelerate time to results Increased number of Increased complexity of samples samples Process more samples Need for complementary with decreasing number characterization methods of resources Asked to do more with less Asked to know more with less 10

What Can Be Accomplished With The Thermo Scientific Vanquish Duo for Dual LC? “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 Finished in half the time Efficient characterization 11

Key Benefits – Thermo Scientific Vanquish Duo for Dual LC Easy-to-use as any other Vanquish system 1. 2. 3. Simple instrument selection Easy method setup with method wizard Automatic method translation 12

Key Benefits – Vanquish Duo for Dual LC No additional bench space needed Vanquish Duo for Dual LC system Vanquish system 55 cm 55 cm 13

Key Benefits – Vanquish Duo for Dual LC Lower costs per sample Cost of ownership calculator: Throughput increase Cost per sample Number of Samples Per Day 192 Throughput gain Doubled 96 throughput and lower costs per sample Single Dual Aggregate analysis experimental conditions 14

Thermo Scientific Vanquish Duo for Tandem LC/LC-MS 15

Analytical Challenge – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS High investments in modern instrumentation Not fully utilizing instrumentation Time not acquiring meaningful data Asked to maximze the instrument outcome 16

Key Benefits – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS Increased Throughput LC-MS Peptide Mapping Example 1.6e9 Right column (n=5) counts 0 counts Left column (n=5) “ Since installing the Tandem LC-MS system we have doubled our min 1.6e9 5.0 10.0 15.0 20.0 25.0 30.0 35.0 throughput and increased the precision of our data !” Throughput increase by Tandem LC/LC-MS THROUGHPUT INCREASE BY (%) 90 Vanquish Duo for Tandem 80 70 Abs. RT Shift Rel. RT Shift Richard Rogers, Just Biotherapeutics 60 Column left to right Column left to right RT RSD [%] Area RSD [%] 50 [min] [%] n=10 n=10 40 30 20 0.023 0.18 0.11 2.47 10 0 2.1 mm - 50 mm 2.1 mm - 150 mm 2.1 mm - 250 mm 5 min Gradient 15 min Gradient 30 min Gradient 17

Key Benefits – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS Advanced column wash without sacrificing throughput 100 Eluent B [%] Standard gradient profile 50 0 0 10 20 30 40 50 60 100 Run Time [min] Eluent B [%] 50 Tandem mode allows for time savings Time savings 0 0 10 20 30 40 50 60 Run Time [min] Additional column wash 100 Eluent B [%] 50 Extra column washing steps without Time savings sacrificing throughput 0 0 10 20 30 40 50 60 Run Time [min] 18

Key Benefits – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS Easy to implement 1. 2. 3. Automatic instrument recognition and fluidic knowledge Standard instrument method Automatic method setup for setup analytical and reconditioning pump 19

Key Benefits – Thermo Scientific Vanquish Duo for Tandem LC/LC-MS Lower costs per sample Cost of ownership calculator: Throughput increase Cost per sample Number of Samples Per Day 48 Throughput and 31 costs comparison 24 between standard single and Vanquish Duo workflows Single Dual Tandem Long Gradient experimental conditions 20

Thermo Scientific Vanquish Duo for Inverse Gradient 21

Analytical Challenge – Thermo Scientific Vanquish Duo for Inverse Gradient Need to detect any compound in a sample, even without chromophore Individual standards for each compound required for quantitative measurements Asked to improve detection and quantification capabilities 22

Unique Detectors: Charged Aerosol Detection • Detection of substances without a chromophore See what others are missing • Consistent analyte response independent of chemical structure • Dynamic range up to four orders of magnitude and highest sensitivity • Relative quantification using virtually any standard Comparison of Charged Aerosol Detection to UV and MS Saving costs for additional detectors or assays 23

Inverse Gradient Compensation for Uniform Response Conventional Gradient Elution Analytical Gradient 85% Eluent at detector Identical mass o.c. Eluent dependent analyte response 15% 15% Analytical Gradient 85% Inverse Gradient Compensation Eluent at detector 15% 15% 50% 85% 85% Inverse Gradient 15% 24

Key Benefits – Thermo Scientific Vanquish Duo for Inverse Gradient Standard free quantification With Inverse Gradient Compensation Without gradient Compensation 0,4 Adenine 0,4 Adenine Tenofovir Tenofovir Emtricitabin 0,3 Peak Area (pA*min) Peak Area (pA*min) 0,3 Tenofovir Disoproxil Emtricitabine 0,2 Tenofovir Disoproxil 0,2 0,1 0,1 0 0,0 0 10 20 30 40 50 0 10 20 30 40 50 Mass on Column (ng) Mass on Column (ng) Similar response curves for all analytes after gradient compensation 25

Key Benefits – Thermo Scientific Vanquish Duo for Inverse Gradient Easy to implement 1. 2. 3. Automatic instrument recognition and fluidic knowledge Standard instrument method setup Automatic method setup for compensation gradient 26