ACQUITY Systems with 2D Technology

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1 ACQUITY Systems with 2D Technology MS Technology Day Sesión Alimentación y Medio Ambiente Barcelona, 21 de marzo de Waters Corporation 1

2 Why Bother with 2D Chromatography? 2012 Waters Corporation 2

3 Two Dimensional Chromatography Principal Separation Method #2 Separation Method # Waters Corporation 3

4 ACQUITY UPLC Systems with 2D Technology Solves Four Key Customer Challenges Increase Selectivity & Sensitivity Eliminate Unwanted Interferences Characterize the Most Complex Samples Mobile Phase Flexibility 2012 Waters Corporation 4

5 Agenda Waters Two Dimensional UPLC Capability Two Dimensional Functions and Analytical Benefits Trapping Heart Cutting Parallel Column Regeneration At-Column Dilution 2012 Waters Corporation 5

6 ACQUITY Systems with 2D Technology Simple to Use Easy to Implement Proven UPLC benefits Resolution Sensitivity Productivity Compatible with a wide range of Chemistries 2012 Waters Corporation 6

7 ACQUITY UPLC Column Manager Designed for 2D Operation Two Easy-to-access, 6-port 2-position valves Two independent temperature controlled zones Low Volume Active Pre-Heaters Ensure best Chromatographic performance run to run and system to system 2012 Waters Corporation 7

8 ACQUITY Systems with 2D Technology Software Control through MassLynx Binary Solvent Manager Quaternary Solvent Manager 2012 Waters Corporation 8

9 ACQUITY Systems with 2D Technology Software Control through MassLynx Simple Control of Valves by Timed Events in Column Manager 2012 Waters Corporation 9

10 Trapping 2012 Waters Corporation 10

11 Increase Sample Loading and Sensitivity with Trap and Transfer Configuration ul Clozapine 1 ppb 2D 3 min gradient MRM of 6 Channels ES+ 327 > e6 % 250 ul Clozapine 1 ppb 1D 3 min gradient Peak distortion Volume Overload 20 ul Clozapine 1 ppb 1D 3 min gradient Waters Corporation 15 Time

12 Heart Cutting 2012 Waters Corporation 16

13 ACQUITY UPLC and Xevo TQ MS Bioanalysis 1D Separation 2012 Waters Corporation 17

14 Running 1D 1D column BSM 1 +SM BSM2 1 BSM 2 2D column Waste Trapping Column 2012 Waters Corporation 18

15 Heart Cutting ACQUITY UPLC with 2D Technology and Xevo TQ MS 1D Separation 2D Separation 2012 Waters Corporation 21

16 ACQUITY UPLC with 2D Technology Impurity Profiling Heart Cut Heart Cut 2012 Waters Corporation 22

17 2D Heart-cutting Benefits Orthogonality In mathematics, two vectors are orthogonal if they are perpendicular, i.e., they form a right angle. The word comes from the Greek ὀρθός (orthos), meaning "straight", and γωνία (gonia), meaning "angle". For example, a subway and the street above, although they do not physically intersect, are orthogonal if they cross at a right angle. Specificity of a method Enhanced sensitivity in MS by dramatic reduction of matrix effect. Possibility to load more and refocused the 1D into the 2D 2012 Waters Corporation 23

18 1D Chromatogram Connect Waste tubing to MS Leave the valve in position 1 to see the 1D peak eluting This is the 1D peak we will have to trap 2012 Waters Corporation 24

19 Separation is performed on the 2D column 2012 Waters Corporation 25

20 2D on the Xevo TQ-S 5pg/ml Plasma Estradiol 2012 Waters Corporation 26

21 1D vs 2D 1D 2D 2012 Waters Corporation 27

22 Parallel Column Regeneration 2012 Waters Corporation 28

23 Parallel Column Regeneration Increase Overall Speed of Analysis Analytical Separation Performed while 2 nd Column is Regenerated 2012 Waters Corporation 29

24 Parallel Column Regeneration Case Study Complex Samples Requiring Long Run times of 35 minutes Regeneration and Requilibration requiring 15 minutes 24 Hour Operation Samples Run 1 Dimensional ACQUITY UPLC 28 ACQUITY UPLC with 2D Technology 41 30% Increase in Throughput 2012 Waters Corporation 30

25 Summary ACQUITY Systems with 2D Technology Simple to Use Easy to Implement Straightforward to operate in various 2D functions Trapping Heart Cutting Parallel Column Regeneration At-Column Dilution 2012 Waters Corporation 37

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