Automating SPE Extractions with the HT400E System by William Amoyal Disruptive Technologies

Disruptive Technologies history Distribution company founded in 2005 Based in Villecresnes, France Cover France, Belgium, Switzerland and other countries depending on product range Agilent Service Partner for UV/Vis spectrophotometry, HPLC and CE 2 people William Amoyal, Sales Manager Franck Do Vale, Service Manager

Disruptive Technologies product range Sample Preparation Nucleic acides, protein and small molecules extraction from hard-to-lyse tissue samples Online SPE system (couples with LCMS) Dissolution/Formulation Dissolution baths, friability and disintegration instruments Physico Chemistry HT Log P and pKa analyzer HT oligonucleotides purity analyzer HT proteins analyzer HT DNA analyzer HT Chiral analyzer Kinetic solubility instruments Service Agilent Channel Partner to service their range of HPLC, UV Spectroscopy and CE Rapid microbiology with Micro PRO Analytical/preparative OPLC chromatography solutions for semi preparative applications (OPLC systems, pumps, sample applicator, video imaging and densitometry instruments, reagent sprayer. Spotter for MALDI and tissue MALDI imaging ELSD detectors Type-C silica hydride HPLC columns Flat sorbent beds for OPLC Accessories and consumables for CE and HPLC Validation kits for HPLC systems

Benefits of HT400E system Reliable Reproducible Flexible User friendly Based on reliable technology of HTA autosampler already used by Agilent, Shimadzu, Perkin etc ... In their GC systems Reproducible Very low CV% inter-assay <2,5% Flexible Accepts many types of SPE cartridge, sample and target vials User friendly Intuitive “drag-and-drop” software Competitive Price Highest cost-to-performance SPE system available on the market

HT400E system with cooling unit

Main features of HT400E system Can be used as an SPE system or HPLC autosampler 20/29 samples max per tray 4 solvents available (more on request – max 11) Syringe and needle washing Software manager QUICK-PREP Sample tray Peltier cooling unit (optional)

Sample tray configurations 20/29 samples per TRAY Sample volume: ∞ ml SPE 1ml , 3ml standard 6 ml optional Target Volume: 2 ml 4 ml Syringe: 1 ml 2.5 ml Other on request

Ideal workflow Cartridge activation Cartridge drying Sample loading Volume, n. of solvent, n. of cycles Cartridge drying # of cycles Sample loading Volume, # of cycles Sample washing Volume, # of solvent, # of cycles Elution Volume, # of solvent, # of cycles, different target vials Reagent addition Volume, # of reagent, different target vials Washing syringe and needle Post-washing cartridge to re-use it

Easy-to-use Quick-Prep software Drag-and-drop software Tools such as Draw, Dispense, Mixing, Elution, Drying, Cartridge Picking, Cartridge Carry, Injection, Valve Washing, Cartridge Conditioning etc ... can be dragged and dropped in method making it possible to write methods in minutes Easy and intuitive software Easy set-up of the workstation (WorkTable) Easy-write work-up process (WorkFlow) Possibility to link methods

Application examples and results The following evaluation has been made using the VMA kit of Chromsystems The procedure has been as requested by Chromsystems protocol

Chromatogramme of VMA separation

Reproducibility of VMA separation Retention Time Area Name ISTD concentration Units 2,685 2177549 VMA 4,670 mg/L 3,680 3811619 iso-VMA 0,000 10,478 4300813 5-HIAA 5,066 13,837 3194021 HVA 4,826   1 2 3 4 5 6 7 8 AVR. S.D. R.S.D VMA 4,67 4,87 4,64 4,58 4,92 4,78 4,73 0,117 2,47% 5-HIAA 5,05 4,98 4,72 4,91 4,82 4,81 5,06 0,123 2,50% HVA 4,83 4,66 4,74 4,75 0,083 1,75%

Carry-over on pathological control Retention Time Area Name ISTD concentration Units 2,585 6645979 VMA 14,150 mg/L 3,500 3968477 iso_VMA 0,000 9,050 9851053 5-HIAA 15,390 12,172 7550284 HVA 12,711 Pathological Control

Carry-over on water WATER Retention Time Area Name ISTD concentration Units 2,585 6645979 VMA 14,150 mg/L 3,500 3968477 iso_VMA 0,000 9,050 9851053 5-HIAA 15,390 12,172 7550284 HVA 12,711 WATER

Specifications HPLC Sample volume: Fixed loop Syringe sizes: 0.5ml, 1ml, 2.5ml, 10ml (2.5ml as standard) Derivatization time: User defined Derivatization function: Choice of reagents Needle wash: With solvent/sample Loop wash: HPLC tray option: 110 sample positions SPE SPE cartridge volumes available: 1ml, 3ml Tray capacity: 29 tubes (20 as standard) Tray temperature control option: Peltier system. 10°C below ambient to +40°C Solvent lines: Up to 10 (4 as standard), plus one wash solvent Physical Characteristics Dimensions (W x H x D): 510 x 735 x 400mm Mass: 15.0kg Power: 110/220V AC, 50/60Hz

For more information on HT400E system To see a video, click on picture besides in Diaporama mode or copy following URL in your browser http://www.youtube.com/watch?v=XApeb60kGUs To download a brochure click on or copy following URL HT400E SPE system brochure http://wamoyal.free.fr/LabHut/SPE/HT400E%20SPE%20system.pdf

Contact details at Disruptive Technologies William Amoyal, Sales Manager Disruptive Technologies 3 Allée des Camélias 94440 Villecresnes Mobile: +33 (0)6 98 64 98 81 Email: wamoyal@disruptechno.com Web: www.disruptechno.com