For purification/separation of organic and aqueous phases from a two phase mixture within a 96 or 24 array tube system

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1 Lollipop Phase Separator For purification/separation of organic and aqueous phases from a two phase mixture within a 96 or 24 array tube system Lollipop Phase Separator GreenHouse Parallel Synthesiser Full Parallel System Integration Genevac Evaporator Stacker Parallel Purification

2 1 The Lollipop is designed for quick and easy purification post synthesis using phase separation The unique design of the Lollipop allows a single 96 pin support plate to be used for the separation of phases in both 96 and 24 array tube systems. The principle of the Lollipop Phase Separator is based on the rapid freezing of the aqueous phase in a two phase mixture, post synthesis, within either a 96 array of 1.2ml tubes or 24 array of 7ml pots. Using chemically resistant peek pins in a micro titer plate format, the aqueous phase freezes to the pins, forming frozen lollipops, allowing for their total separation from the organic liquid phase. Lollipop Summary Ideal for the post synthesis purification of mixtures in combinatorial chemistry and parallel synthesis processes Innovative and cost-effective alternative to robotic systems Compatible with 96 or 24 well micro titer plate format for subsequent storage or screening Can be used with both organic phases that are more dense (i.e chloroform) or less dense (i.e ethyl acetate) than aqueous Based on the rapid freezing of the aqueous phase in a two phase mixture, with the aqueous phase being removed as a frozen lollipop Takes less 15 minutes to process 96 samples Sample tubes are easily transferred from their racks to the appropriate Lollipop tube support frame Anodised extraction stand for chemical resistance 96 array of chemically resistant Peek pins for 24 or 96 well formats Support frame compatible with Genevac Vacuum Centrifuges An effective and economic alternative to expensive robotic sample handlers. Components present prior to synthesis Components present at the end of the reaction Add dilute aqueous acid to mixture.mix aqueous and organic phases and then allow to separate. Add dilute aqueous base to mixture. Mix aqueous and organic phase and then allow to separate. Purified target mixture after two Lollipop phase separations Carboxylic Acid Reagent 1 Amine Reagent 2 Catalyst Carbodimide Catalyst Dimethyl formamide Reaction Combinatorial Synthesis Impurity Soluble in Aqueous Base Urea Impurity Soluble in Aqueous Acid Dimethyl formamide This solvent is miscible with water and organic solvents and must therefore be removed by vacuum centrifugation or similar Dissolve mixture in non-miscible solvent Impurity Soluble in Aqueous Base Urea Impurity Soluble in Aqueous Acid Perform First Lollipop Phase Separation Impurity Soluble in Aqueous Base Remove frozen Aqueous impurity on PEEK pin Perform Second Lollipop Phase Separation Remove frozen Aqueous impurity on PEEK pin Note: Prior to Purification Removal of the reaction solvent by vacuum centrifugation will be required if a miscible organic solvent is used i.e. dimethyl formamide or acetonitrile. (The Lollipop tube support frames are designed to fit into a Genevac vacuum centrifuge.) The dried reaction mixture is then dissolved in a non-miscible organic solvent i.e. ethyl acetate or chloroform. Where a non-miscible organic solvent such as dichloromethane is used for the reaction; Lollipop purification can be done straight way.

3 2 The Lollipop Phase Separator provides an effective and economic alternative to the use of expensive robotic sample handlers for purification after synthesis, using phase separation. The Lollipop Phase Separator uses either 1.2ml PP tubes or 7ml HDPE pots with tapered sides for easy aqueous phase extraction If the target sample is in the organic phase, then the frozen lollipops can be washed away with warm running water If the target sample is in the aqueous phase, then the frozen lollipops are allowed to defrost into a 96 or 24 well micro titer plate min Sample tubes are easily transferred from their racks to the appropriate Lollipop tube support frame 2 9 The pin support plate with the frozen aqueous Lollipops is then removed. Dependent on the chemistry, the target sample may be in the frozen aqueous or organic liquid phase The organic and aqueous phases are dispensed into the individual tube wells using a standard multichannel pipettor. The phases can be pre-mixed before transfer or mixed in situ mins 8 >1 min The pin support plate is then separated from the tube frame by lowering the Lollipop stand jack. Thus extracting the frozen aqueous Lollipops from the organic liquid phase, which remains in the tubes >1min >1 min 2-3 mins The complete assembly is removed from the acetone bath and transferred to the lollipop extraction stand The 96-pin support plate is placed into the tube frame so that the pins are immersed in the two-phase solution. The 96 pins will also work with the 24 well format Dry ice (CO2) is added to a bath of acetone until the temperature is reduced to around -25 C to -30 C Complete tube frame and pin support assembly is placed in the acetone bath. Bath temperature should be maintained at around -25 C until the aqueous phase is frozen The aqueous phase freezes to form a lollipop on each pin, leaving the organic solvent as a liquid. 4-6 mins - for 24 well or 2-5 mins - for 96 well Important: Purification Notes The Lollipop Phase Separator can be used with both organic phases that are more dense (i.e. chloroform) or less dense (i.e. ethyl acetate) than aqueous. The Lollipop purification procedure may be repeated up to 3 times with an aqueous acid, aqueous base and a neutral aqueous phase if the chemistry requires. Ideal volumes for the 1.2ml x 96 array are 300µl organic phase and 300µl aqueous phase per tube. Ideal volumes for the 7ml x 24 array are 2ml organic phase and 2ml aqueous phase per pot.

4 3 Lollipop - Full integration with parallel synthesis, parallel evaporation and parallel purification systems, such as the GreenHouse, Genevac Vacuum Centrifuges and Stacker. It is essential to provide parallel solutions to post synthesis procedures to avoid downstream bottle-necks... The Lollipop is designed to optimise integration with 96 and 24 well MTP (micro titer plate) formats. This greatly speeds up the transfer of samples between synthesis, separation, purification and evaporation stages. MTP format allows easy transfer of samples using standard format multichannel pipettors and liquid handling systems (see overleaf). Reduced handling of individual tubes, avoiding the risk of confusion. Full integration with the RDT GreenHouse for the parallel purification of samples in a standard 24 position MTP footprint. Using a multi-channel pipettor you can quickly transfer samples from the GreenHouse to the Lollipop post synthesis, for parallel and sequential work-up of all 24 reactions via phase separation. Transfer samples from the Lollipop tubes to the RDT Stacker for further SPE and Filtration. The Lollipop support frames are also designed to fit directly into the complete range of Genevac vacuum centrifuges. Allowing the user the ability to transfer directly from reactor to the evaporator without the hassle of reloading reaction tubes or the risk of mixing up samples. No Are solids present? Is an aqueous workup necessary? Lollipop Phase Separator GreenHouse Parallel Synthesiser Is solvent miscible with water? No Is solvent more dense than water? Add non-water miscible solvent & water Genevac Evaporator Using a multi-channel pipettor you can quickly transfer samples from the GreenHouse to the Lollipop post synthesis, or to the Stacker for further SPE and/or filtration steps. Stacker Parallel Purification System No Phase Sep Stacker Filtration SPE Pure Product GreenHouse Reaction Block and Genevac Vacuum Centrifuge

5 4 Lollipop Phase Separator For the purification/separation of organic and aqueous phases from a two phase mixture International patents pending All aluminium construction with hardened blue anodised surface for improved chemical resistance 96 array of chemically resistant PEEK pins for 24 or 96 well formats Removable 96 and 24 tube support frames are compatible with Genevac Vacuum Centrifuges The aqueous phase is frozen to the peeks pins to form a lollipop, leaving the organic solvent as liquid Lollipop accommodates both 24 x 7ml pots and 96 x 1.2ml tubes Rapid - takes less than 15 minutes to process 96 samples The Lollipop can be used with both organic phases that are more dense (i.e. chloroform) or less dense (i.e. ethyl acetate) than aqueous. Lollipop stand has integral jack to separate pin support plate and frozen lollipops from the tube support frame after freezing. Developed by medicinal chemists at GlaxoSmithKline Medicines Research Centre and engineers from GlaxoSmithKline s UK Applied Technology department.

6 Impact 2 6 Channel Electronic Pipettor The cordless, rechargeable Impact 2 pipettor accepts six extra-long, narrow 1250µl pipette tips, allowing you to reach to the very bottom of longer tubes or deep wells (including GreenHouse Reaction Tubes). The Impact 2 allows programming and storage of five different protocols of up to 40 steps each, with one additional floating program. Plus 3 standard and 2 additional slow speed controls for working with viscous liquids. The Impact 2 s unique expandable tip spacing system allows transfer of samples from 96 to 24 well formats with ease (freely expandable from 9 to 19.81mm). The Impact 2 pipettor is available in other volumes and with 8 or 12 channels - details on request. Cat No. Description Qty Per Pack RR98001 Lollipop Phase Separator 1 (includes 96 and 24 Place Tube Support Frames) Lollipop Spare Parts RR Place Tube Support Frame 1 RR Place Tube Support Frame 1 RR98083 Clips for Tube Support Frame 2 RR Pin Support Plate (without Pins) 1 RR Pin Support Plate with 96 Peek Pins 1 RR98090 Replacement Peek Pins, Loose x 1.2ml Polypropylene Tube System RR x 1.2ml PP Racked Tube System (10 x 96) 960 RR98011 Loose 1.2ml PP Tubes 1000 RR98012 Cap Strips for 1.2ml Tubes, (120 strips of 8) x 7ml HDPE Pot System RR98020 Loose 7ml HDPE Pots 100 RR Place Polycarbonate Pot Holder with Cover 1 RR98022 Extendable Storage Holder for RR Other Accessories RR98023 Borosilicate Glass Freezing Tray 1 RR98024 Protective Cold Temperature Gloves Pair RR98025 Micro-Mix Programmable Micro Plate Shaker 1 RR98028 Digital Thermometer & Probe, -50 C to 150 C 1 24 Well Micro Titer Collection Plates T-9000U Titan 24 x 9ml Conical Bottomed PTFE Plate 1 RR ml x 24 Well Clear Polypropylene Micro Plate 25 Impact Multi-Channel Pipettors RR70048 Impact 2 6 x 1250ul Expandable Multi-Channel Pipettor 1 RR70050 Impact 2 8 x 1250ul Expandable Multi-Channel Pipettor 1 RR ul Extra-long Bulk Pipette Tips 500 RR ul Extra-long Racked Pipette Tips 720 RR70051 Impact 2 Pipettor Stand 1 Please specify preferred voltage when ordering. Other Impact Pipettor Styles also available on request Micro-Mix Micro Plate Shaker A fully programmable shaker with an extremely low profile. Ideal for the mixing of aqueous and organic phases within the Lollipop racks prior to phase separation. Holds up to four standard micro titer plates or similar. GreenHouse Parallel Synthesiser Stacker Parallel Purification System 24 position personal synthesiser for parallel solution phase and solid supported reagent based chemistry A 24 position personal parallel synthesiser designed by medicinal chemists at GlaxoSmithKline Rapid sequential and parallel purification in a 24 well micro titer plate footprint using standard filtration, phase separation and SPE columns... A Parallel Purification system for the purification/separation of organic and aqueous phases from a two phase mixture Radleys Discovery Technologies are specialists in combi-chem and parallel chemistry... As a dynamic organisation Radleys Discovery Technologies are best able to react to the needs of this constantly developing market sector. Radleys Discovery Technologies specific areas of expertise is focused on parallel synthesis and parallel purification consumables and apparatus. The essential products that are the cornerstone of your parallel chemistry program. Radleys Discovery Technologies 2001 Radleys Discovery Technologies, Shire Hill, Saffron Walden, Essex, CB11 3AZ. United Kingdom. Tel: 44-(0) Fax: 44-(0) sales@radleys.co.uk Web:

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