Instructionsforuse. Speedisk Column Instructions. Introduction. Application

Transcription

1 Instructionsforuse Speedisk Column Instructions Introduction Speedisk columns are disposable, ultra-clean, polypropylene columns which are pre-packed with high performance microparticles and uniquely configured, as shown on the right. The Speedisk configuration shortens run times, increases capacity, and may eliminate pre-filtration and evaporation steps. Columns are available in 1 ml, 3 ml or 6 ml sizes, and there are more than 25 BAKERBOND silica sorbents and Baker Polymer Resins to choose from. BAKERBOND chemistry brings the traditional advantages of bonded silica to SPE and, in addition, provides lot-to-lot reproducibil-ity, and highly stable particles for use over the ph range New Baker hydrophilic and hydrophobic polymer resins are highly rigid, have large surface areas, and are stable over ph range In addi-tion, they are ultra-clean for use in preparation of samples for advanced detection methods. Speedisk column s newly designed, luer tipped tube fits into both a traditional manual vacuum processor and the novel Speedisk 48 Posi-tive Pressure Processor. (The manual positive pressure processor enables easier, finer flow rate control, and a more uniform flow rate from column to column than traditional manual vacuum processors.) The tube inlet is round-rimmed with a small flange. A rimless 1 ml tube is also available for use in a 96-position tube rack that has a micro-plate footprint. In addition, tube inlet adapters are available for use with automated liquid delivery systems that use injectors and control flow rate with pressurized gas. Please note that Speedisk columns are operated with smaller solvent volumes and have higher capacity per milli-gram sorbent than the conventional SPE columns to which you may be accustomed. Please refer to Tables 3, 4 and 5 for operating guidelines. Application Speedisk columns are used for sample preparation before analysis by research, clinical, and environmental chemists preparing to perform HPLC, LC, GC or other high resolution analytical methods. Sample enrichment or clean up with Speedisk columns enables low detection limits. INSTRUCTION FOR USE

2 Speedisk Column Characteristics Tube Guard Filter BAKERBOND Silica Sorbent Baker Polymer Resin Packaging Typical Recovery 80-95% Extraction Run Time Two inlet types (flanged/round rimmed for 1, 3, 6 ml tube, and rimless for 1 ml tube) and 3 tube volumes Male luer Flat bottom to reduce holdup volume and increase final concentration Ultra-clean polypropylene composition Removes suspended solids and may eliminate pre-extraction step Promotes high flow rates Low resistance to flow for easy liquid displacement or drying Ultra-clean Ultra-clean Base particle 10 micron HPLC silica Ultra-Stable Silica Micro-particle Bonding Technology Capacity/mg sorbent Increased relative to conventional SPE sorbent due to high available surface area of micro-particle Operating ph range 2 10 Base particle 15 micron for both hydrophilic polymerand hydrophobic polymer Operating ph range 1 14 Using Ultra-clean Polymer Micro-particle Technology Capacity/mg polymer resin Increased relative to conventional columns because of high available surface area of micro-particle Air and moisture tight Laminar construction and use of micro-particles results in bed and solvent volume reduction and flow rate increase. Extraction run time is shortened to as little as 11% that of columns using 40 micron or larger particles. Reduced final volume results in higher sample concentration and may eliminate a post-extraction evaporation step. Product Range and Selection There are more than 25 BAKERBOND sorbents from which to choose, including silica-based sorbents with 60 Å or 275 Å pore size for normal phase, reversed phase, hydrophobic interaction and ion exchange separations, and hydrophobic and hydrophilic polymer resin particles with 200 Å pore size for adsorption, reversed phase and ion exchange chromatography. Column sorbent types and part numbers are found in Tables 1 and 2, and column tube volume and bed mass are selected from Table 3. Table 1 Speedisk Polymer Column Sorbent Selection Baker Resin Functional Mode Pore Size a General Applications Speedisk Column Part # H 2 O-Philic DVB Adsorption 200 Å Polar to non-polar analytes 8108 H 2 O-Philic SC-DVB Cation Exchange (strong) 200 Å Ionic, basic analytes 8111 H 2 O-Phobic DVB Adsorption 200 Å Slightly polar to non-polar analytes 8109 H 2 O-Phobic SC-DV Cation Exchange (Strong) 200 Å Ionic, basic analytes 8196 H 2 O-Phobic SA-DVB Anion Exchange (Strong) 200 Å Ionic, acidic analytes 8113 H 2 O-Phobic WA-DVB Anion Exchange (Weak) 200 Å Ionic, acidic analytes 8115 a. by Mercury Porosimetry INSTRUCTIONS FOR USE 2

3 Table 2 Speedisk Silica Column Sorbent Selection and Part Number BAKERBOND Silica Functional Mode Pore Size General Applications Speedisk Column Part # Octadecyl C 18 RP 60 Å Non-ionic, non-polar to moderately polar analytes 7606 Octadecyl LightLoad C 18 RP 60 Å Non-ionic, non-polar to polar analytes 8151 PolarPlus Octadecyl C 18 RP 60 Å Non-ionic, basic, non-polar to polar analytes 8153 Octyl C 8 RP 60 Å Non-ionic, non-polar to moderately polar analytes 8154 PolarPlus Octyl C 8 RP 60 Å Non-ionic, basic, non-polar to moderately polar analytes 8156 Ethyl C 2 RP 60 Å Non-ionic, highly hydrophobic analytes 8157 Cyano CN NP 60 Å Non-ionic, non-polar, aromatic analytes (Synthetic DNA) 8159 Phenyl C 6 H 5 RP 60 Å Polar from non-polar/polar solvents using hydrogen bonding like mechanisms Silica NP 60 Å Separate polar from non-polar solvents 8163 Amino NH 2 NP 60 Å Lipids (fatty acids, cholesterol) 8165 Diol COHCOH NP 60 Å Strongly polar analytes 8167 Quaternary Amine N + Anion Exchange (strong) 60 Å Ionic, acidic analytes 8168 Aromatic Sulfonic Acid Cation Exchange (strong) 60 Å Ionic, basic analytes 8170 Carboxyl Acid Cation Exchange (weak) 60 Å Ionic, basic analytes 8172 Narc -1 Mixed 60 Å Carboxy-tetrahydrocannabinol (THC) 8174 Narc-2 60 Å Hydrophobic/basic analytes (Cocaine, Benzoylecgonine) 8175 Butyl C 4 RP 60 Å Non-polar 8184 CBx WP IX 275 Å Proteins/antibodies/hemoglobin 8176 PEI WP IX (Weak) 275 Å Proteins/antibodies 8178 Butyl WP HIC 275 Å Proteins/antibodies, desalting 8180 HI-Propyl C 3 HIC 275 Å Proteins/antibodies, desalting 8182 Table 3 Speedisk Column Size and Part Number Suffix Selection Part Number Suffix Format Sorbent mg/column Columns/pack -01 a Round rimmed, 1 ml column Round rimmed, 1 ml column Round rimmed, 3 ml column Round rimmed, 3 ml column Round rimmed, 3 ml column Round rimmed, 6 ml column Round rimmed, 6 ml column Round rimmed, 6 ml column ml rimless column Speedisk 96 Column (microplate) a.use the part numbers in Table 2 in conjunction with suffix in this table. Example: Octadecyl C18 would be for a 1 ml, 20 mg column;or for a 6 ml, 50 mg column, etc INSTRUCTIONS FOR USE 3

4 Performing Solid Phase Extraction Solid phase extraction is typically performed as follows: 1. ConditioningColumn conditioning with an appropriate solvent to solvate the functional groups of the sorbent. 2. Sample ApplicationSample Applica-tion to promote interaction of analytes with the functional groups on the sorbent and retain the analytes on the column. 3. WashingColumn washing with an appropriate solvent to elute impurities while leaving the analytes on the column. 4. ElutionAnalyte elution with a strong enough solvent to displace the analyte from the sorbent. For additional information, please refer to Solid Phase Extraction: A technical guide to theory, method development and use, J.T. Baker literature #8008. Selection and Use of Speedisk Columns After you select the appropriate column sorbent (see Tables 1 and 2) and column size (see Table 3), consult Table 4 for ranges of column capacity and examples of operating conditions. Table 4 Example Speedisk Column Capacity, Flow Rate and Extraction Volume Speedisk Column Bupivacaine Capacity a,1 ml, 20 mg Column Bupivacaine Capacity a,3 ml, 60 mg Column Bupivacaine Capacity a,6 ml, 100 mg Column mg/mg Total mg mg/mg Total mg/mg Total C H 2 O-Phobic DVB H 2 O-Philic DVB SPE Steps Flow Rate ml/min Flow Rate ml/min Flow Rate ml/min Load Wash Elute Speedisk Column Final Extract Volume (µl),1 ml Column Final Extract Volume (µl),3 ml Column Final Extract Volume (µl),6 ml Column C mg bed mg bed mg bed mg bed mg bed mg bed H 2 O-Phobic DVB 20 mg bed mg bed mg bed H 2 O-Philic DVB 35 mg bed mg bed mg bed mg bed mg bed mg bed mg bed mg bed mg bed a.capacity (mg) at breakthrough of 1 mg sorbent for Bupivacaine in 0.2M phosphate buffer, ph 6.0. INSTRUCTIONS FOR USE 4

5 Speedisk Column and microparticle performance can be related to conventional SPE column and large particle performance as follows: The relative dynamic binding capacities of two sorbents of different particle diameter, identical pore structure and functionality, and similar surface coverage is directly related to the quantity of available binding sites. Typically, 10µ bonded silica particles exhibit two to three times the dynamic capacity (per mg sorbent) of 40µ particles. (See Table 4) A maximum effective operating flow rate for an SPE column is one which allows sufficient time for mass transfer and interaction between analyte and functional group. For two sorbents of different particle diameter, but identical pore structure and surface coverage, the flow rate maximum is inversely related to particle diameter. For example, a Speedisk column filled with 10µ particle has a maximum effective flow rate that is as much as 300% higher than an SPE column filled with 40µ particles. (See Table 4) By affecting sorbent capacity/mg and column bed volume, particle diameter also determines effective column operating volumes (conditioning, wash and elution volumes). These volumes will, in turn, affect run time and final concentration. For example, a Speedisk column filled with 10µ particles has minimum effective operating volumes that are 33% of that of an SPE column filled with 40µ particles. (See Table 4) In summary, higher flow rates (up to 300%) and reduced volumes (as little as 1/3) lead to dramatically faster run rates. For example, a Speedisk column filled with 10µ particles may be run 9 times faster than an SPE column of equivalent capacity filled with 40µ particles. Application Examples and Results Extraction of Caffeine from Porcine Serum by Speedisk Column Octacedyl C18 ( *, 10 mg) Procedure 1. Spike 1 ml of serum with 50 µg of caffeine. 2. Place column on processor and condition with 200 µl methanol followed by 100 µl of water. Keep column wet until next step. 3. Add sample to column at a rate of 1 2 ml/min. 4. Wash column with µl of water. This volume may be increased to µl, if there are excessive impurities. 5. Elute with µl methanol. Bring extract to 1 ml with HPLC mobile phase solvent. Analyze by HPLC. 6. Recovery = 95%. * µg and µg of caffeine can be extracted by and , respectively. Extraction of Clonazepam from Human Serum by Speedisk Polymer Column H 2 O-Phobic DVB ( , 20mg) Procedure 1. Spike 0.5 ml of human serum with 200 µg of Clonazepam and dilute to 1 ml with water. 2. Place column on processor and condition with 100 µl of methanol followed by 50 µl of water. 3. Add spiked serum sample and apply vacuum. 4. Wash column with µl water. (For undiluted serum, this volume should be increased to µl because of additional impurities.) 5. Elute with µl acetonitrile, and analyze by HPLC. 6. Recovery = 92 97%. INSTRUCTIONS FOR USE 5

6 Table 5 Related Products, Accessories and Literature Product Name Description/Purpose Product Number Speedisk 48 Positive Pressure Processor (1) 48-position, two-stage processor, (1) waste container, (1) 3 ml SPE column rack, (1) mm collection tube rack, waste bin, gas supply adapter and tubing; operated under 100 lbs N 2 pressure Brochure Number BAKER spe-12g Column Processor Manual, vacuum processing of BAKERBOND spe columns BAKER spe-24g Column Processor Manual, vacuum processing of BAKERBOND spe columns Solid Phase Extraction: a technical guide to theory, method development and use BAKERBOND spe application notes 55-page manual designed to illustrate the effective use of solid phase extraction 8008 SPE of biological, environmental, food, and pharmaceutical samples using BAKERBOND spe and Speedisk products Baker Analyzed HPLC Methanol Extraction/concentration solvent Baker Analyzed HPLC Water Extraction/concentration solvent Baker Analyzed HPLC Acetonitrile Extraction/concentration solvent Storage Conditions Store sealed at room temperature, away from moisture and light. Ordering Information To order Speedisk products, contact any authorized J.T.Baker distributor, or in North America call JTBAKER ( ). In Europe, call Information And Assistance Contact us at or in North America, please call JTBAKER ( ), or for Customer Service. In Europe, please call Central Service, Deventer, Holland at The information contained herein is to our best knowledge true and accurate. Recommendationsand suggestions are made without guarantee of favorable results because conditions of use arebeyond our control. This data shall not be construed as a recommendation to use any product inconflict with existing patents covering any material or use. Phillipsburg, NJ 9001: 2008 & 14001: 2004 Paris, KY 9001: 2008 Mexico City, Mexico 9001: 2008 Deventer, the Netherlands 9001: 2008 & 14001: 2004 & 13485: 2003 Selangor, Malaysia 9001: 2008 About Avantor Performance Materials Avantor Performance Materials manufactures and markets high-performance chemistries and materials around the world under several respected brand names, including the J.T.Baker, Macron Fine Chemicals, Rankem, Diagnova, BeneSphera, and POCH brands. Avantor products are used in a wide range of industries. Our biomedical and life science solutions are used in academic, industry and quality control laboratories for research, pharmaceutical production and medical lab testing, while our electronics solutions are used in the manufacturing of semiconductors and flat panel displays. Based in Center Valley, Pennsylvania (USA), Avantor is owned by an affiliate of New Mountain Capital, LLC. For additional information please visit or follow Ordering Information and Assistance Customer Service and Technical Service toll free: AVANTOR ( ) outside of u.s. tel: fax: info@avantormaterials.com AskAvantor Our Web site features ASK Avantor, which includes live chat capabilities with customer service representatives. Lit Number: Avantor Performance Materials, Inc. All rights reserved. Trademarks are owned by Avantor Performance Materials, Inc. or its affiliates unless otherwise noted. Corporate Headquarters Avantor Performance Materials, Inc Corporate Parkway Suite #200 Center Valley, PA USA Worldwide Locations China Malaysia North America India Mexico Taiwan Korea The Netherlands For contact information at these locations, visit Us/Worldwide-Directory.aspx