The world leader in serving science Alternate Channels Meeting 12 th June 2007 Tom Whitehouse European Commercial Marketing Manager Small Molecules

The world leader in serving science TurboFlow ® & Multiplexing Technology

3 History Founded as a Novel Chromatographic Media Company focused on protein purification Discovered High Turbulence Liquid Chromatography (HTLC™) 6 patents issued Launched development scale preparative instrumentation – Cohesive Introduced pilot scale preparative instrumentation – Cohesive Developed multi-dimensional chromatography (MDLC) – Cohesive Opened UK office Developed Aria OS multiplexing software and instrumentation Established a sales and support agreement with Applied Biosystems Won patent infringement lawsuit against Waters Corporation

4 History European patent reinstated - Established a sales and support agreement with Thermo Electron - Launched 0.5mm ID & ion–exchange HTLC columns First Clinical /Hospital system sold Release Aria OS Launch Cyclone MAX & MCX mixed-mode HTLC columns Launch Transcend™ (High Pressure LC) System - Launch Propel™ analytical columns - First two instruments installed in India - Acquired by Thermo Fisher Scientific Release Aria OS 1.6

5 What is TurboFlow?  Chromatographic technique On-line sample extraction plus chromatographic separation Exploits difference between large and small molecules Incorporates column chemistry  Large particle columns (30 µm or larger)  High flow rate, low back pressure  Efficient mass transfer created by turbulence  Separates low MW analytes from high MW matrix  Allows for the direct injection of biological fluids Eliminates sample prep (SPE, protein precipitation, liq/liq extraction etc) Cleaner extracts, less source maintenance Fast analysis

6 Turbulent Flow With Turbulence Without Turbulence High linear velocities with large particles create turbulence

7 Molecules Interact with the Stationary Phase  Column particle contains small pores  Fluid inside pore is nearly stagnant

8 Small molecules diffuse into pores faster than large molecules Small and Large Molecules Diffuse at Different Rates

9 Large Molecules Flow to Waste  Large molecules do not have time to diffuse into pores  They are flushed through the column by the high-velocity mobile phase  Sample residence = 300 milliseconds Particle Pore

10 Column Chemistry  Small molecules that have diffused into pores either bind to selected column chemistry or exit the column Particle Pore

11 Small Molecules are Eluted A change in mobile phase composition elutes compounds of interest to the detector or to an analytical column Particle Pore

12 TurboFlow ® Demonstration

13 SPE, PPT and TurboFlow ® method SPE TLX (A) Blanks Rat Plasma PPT 4:1 Intensity Time (min) 50:50 MeOH:H 2 0 Buffered H2050:50 MeOH:H20 TLX PPT SPE [ J. L. Herman, Cephalon, Inc., Brandywine Parkway, PA] Ion Suppression Effects

14 Direct Injection of Plasma with TurboFlow ® Method vs. Protein PPT. of Plasma with LC/MS/MS 250 pg/mL Cyproterone Acetate ACN crashed (1:1) plasma injection 250 pg/mL Cyproterone Acetate Direct plasma injection w/ TurboFlow® method Greatly Improved S/N Less Sample Prep

15 Cohesive Customer Applications  Drug Discovery  In Vitro screening  In Vivo screening  Combinatorial chemistry  Drug Development  Preclinical  Clinical  Diagnostics  Therapeutic drug monitoring  Esoteric assays  Biomarkers  Food & Environmental  Forensics & Toxicology

16 Why an Aria TLX System?  Optimized system to maximize the patented TurboFlow ® columns and methods  Unsurpassed expertise in developing methods and supporting systems  Aria ® OS software for system control and communication with MS software.  Comprehensive service and applications support.  Complete Solution!

17 Aria TLX1 HTLC System

18 Aria TLX1 MD HTLC System

19 Multiple Column Module

20 MS “IDLES” for 75% of the Time meaningful data Multiplexing Aria Systems MS Capacity of 15 samples/hour 0 4 min.

21 Parallel System, Serial Data Multiplexing with Aria LX Systems Quadruple the Throughput Pump Column Pump Detector Autosampler

22 Multiplexing your Method MS capacity of 60 samples/hour MS “IDLES” for <4% of the time Sample injections and acquisitions are staggered among four HPLC systems in an intelligent manner min. Acquisition window

23 Increasing sample throughput with Multiplexing The MS “IDLES” for <4% of the time Maximize the use of your MS 04 min. Aria OS controls Serial delivery of samples to MS Each sample is an individual data file 1 2 3

24 Testosterone and Androstenedione 3.4 minute method, data collection for 45 seconds. SYSTEM 1 Testosterone and Androstenedione 3.4 minute method, data collection for 45 seconds. SYSTEM 2 Double the samples analyzed 50uL Plasma injection Transcend TLX2 system

25 Multiplexing with TurboFlow ® L Pump Detector Autosampler Four parallel systems, synchronized by Aria software, all lead into the MS and work independently from each other HPLC TFLC E Pump TFLC E Pump TFLC E Pump TFLC E Pump

26 TLX-2 HTLC System

27 TurboFlow ® on-line extraction Eliminates SPE/LLE/PPT Ultra Fast, Rapid Resolution, UPLC chromatography High resolution, fast separations Multiplexed Ultimate throughput New! Cohesive Transcend ™ System

28 Peaks eluting in 18 minutes vs minutes with the Transcend TLX System. Testosterone, Androstenedione and metabolites. 32 minutes plus equilibration method without sample prep. HPLC Steroids and metabolites HPLC separation 32 minutes without sample prep

29 StartSecFlow%A%BAction Loading sample into TFC column Slow down flow Transfer sample to LC column Analytical Run Wash off column Re-equilibrate for TFC next injections TurboFlow ® and LC/MSMS analysis Transcend™ System 5.5 minutes: Testosterone, Androstenedione, and 8 metabolites.

σ-hydroxytestotsterone β-hydroxytestotsterone σ-hydroxytestotsterone 4. 6-β-hydroxytestotsterone σ-hydroxytestotsterone 6.16-β-hydroxytestotsterone 7. 2-σ-hydroxytestotsterone 8. 2-β-hydroxytestotsterone 9. Androstenedione 10.Testosterone 9 FAST! Steroids and 9 metabolites Cohesive Transcend System

31 10µL 6%BSA 10µL PBS 10µL Rat plasma 5.5 Same TurboFlow ® method from 3 different matrices Save time with method development. Use TurboFlow ® for multiple matrices.

32 Transcend™ TLX 2 System  Optimized system for reliable, reproducible high throughput  Flexible system solves many Bioanalytical challenges  Pressure up to 600BAR for use with <2µ particles  Advanced Aria software for the optimum control of autosampler, pumps and valve interface module  Transcends all other systems Transcend™ TLX-2

33 System Configurations  TurboFlow ® Systems Aria TLX1 Aria TLX1 MD Aria TLX2 Aria TLX4  Multiplex / Parallel LC Systems Aria LX2 Aria LX4  Transcend™ Systems Transcend™ TLX1 Transcend™ TLX2 Transcend™ LX2 Transcend™ LX4

34 Territory Installed Base  Sanofi-Aventis, Hungary - 2 x TLX1  Pharma Serv, Romania - 2 x Transcend LX2  RE & D VUFB s.r.o, Czech Republic – 1 x TLX1

35 Contacts  Tom Whitehouse  Dawn Briggs – Sales Specialist UK, Nordic & Netherlands  Karen King – product information, customer, sales, quotations etc  Padma Sekhar – Application Chemist  Sales Specialist based in Switzerland  Sales Specialist based in France  Factory – Franklin, MA, USA

36 Website

The world leader in serving science Alternate Channels Meeting 12 th June 2007 Tom Whitehouse European Commercial Marketing Manager Small Molecules