SPE. Simply Efficient! Watchers bond Solid Phase Extraction by ISU. Ver
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1 SPE Simply Efficient! Watchers bond Solid Phase Extraction by ISU Ver
2 Solid Phase Extraction Basic principles of Solid Phase Extraction Solid phase extraction (SPE) is a powerful method for sample preparation and is used by most chromatographers today. It has capabilities in a broad range of applications such as environmental analyses, pharmaceutical and bio-chemical analyses, organic chemistry and food analyses. SPE has capabilities in a broad range of applications: Environmental analyses Pharmaceutical and biochemical analyses Organic chemistry Food analysis
3 Watchers bond Advantages of SPE compared to classical liquid-liquid extraction Lower consumption of solvents Faster - enormous time savings Lower costs per sample Potential for automation High consistency in individual sample handling More specific selectivity because of the broad range of adsorbents and different retention mechanisms Optimisation of extraction by variation or adjusting of the solid phase and chromatographic conditions How does SPE work? Retention of the analyte - analyte molecules are enriched on the adsorbent - interfering components and solvent molecules(matrix) are not retained - remaining interfering components are washed from the adsorbent Washing Elution - the analyte is removed from the adsorbent by elution with a suitable solvent Conditioning Moisturising of the solid phase with the matrix Sample application Sample is pressed or drawn through the solid phase Elution Retention of interfering components - analyte molecules show no interaction with the adsorbent - interfering components and solvent molecules (matrix) are retained - analyte molecules are washed from the adsorbent - the solid phase is simply used to filter the sample Since analytes can be either adsorbed on the SPE packing material or directly flow through while the interfering substances are retained, two general separation procedures are possible both cases are shown in the figure above. Comparison of recovery rates for Watchers bond Easy and other SPE phases Compounds investigated: ciprofloxacin (1), doxepin (), cinoxacin (3) Column types: Watchers bond Easy, 500mg, 3ml Oasis, Watchers bond HR-P, Watchers bond C18 ec Column conditioning: a) ml methanol, than ml dist. Water, b)no conditioning Sample application: slowly force or aspirate the sample (100~00μm/compound in 00ml water) through the column Column washing: 10ml water; Elution: slowly aspirate 10ml methanol - THF(1:1, v/v)through the column a) Procedure with conditioning and equilibration b) Procedure without conditioning and equilibration conditioning with methanol conditioning with methanol equilibration with water equilibration with water sample application sample application let the cartridge run dry washing / elution washing / elution Easy Oasis HR-P C18 ec Easy Oasis HR-P C18 ec 3
4 Solid Phase Extraction Cross Reference Reversed Phase Easy HLB Strata -X C18ec C18 C18 C18-E C18 C18-U C18 C8 C8 C8 C8 C8 C 6 ph ph Phenyl Normal Phase SiOH Silica Si-1 Silica Florisil Florisil Florisil FL-PR Florisil NH NH NH NH NH CN CN CN-U CN CN Alox A Alox N Alox B Ion Exchangers SB SAX SAX SAX SAX SA SCX SCX SCX HR-XC MCX X-C HR-XA MAX Aluminium oxid A Aluminium oxid N Aluminium oxid B Selection guide for SPE phases and solvents Sample solubility soluble in water soluble in organic solvents not ionic Solvent aqueous aqueous aqueous organic organic ionic Sample polarity nonpolar moderately polar cationic anionic nonpolar moderately polar polar polar Phases recommended Easy SiOH CN PCA SB Easy SiOH CN for adsorption HR-P NH OH SA NH HR-P NH OH C18 ec PA PSA DMA C18 ec PA C18 DMA PS-H+ PS-OH- C18 DMA C18 Hydra NH C18 Hydra NH C8 C8 C4 C4 C C C 6 C 6 CN CN PS-RP Solvents recommended hexane CHCl 3 CHCl 3 acids bases hexane CHCl 3 CHCl 3 4 for elution(selection) CH Cl CH Cl CH Cl salt salt CH Cl CH Cl CH Cl acetonitrile ethyl ethyl solutions solutions acetonitrile ethyl ethyl alcohols acetate acetate buffers buffers alcohols acetate acetate alcohols alcohols alcohols alcohols water water
5 Watchers bond Molecular interactions in SPE SPE adsorbents are most commonly categorised by the nature of their primary interaction mechanism with the analyte of interest. The three most common extraction mechanisms used in SPE are reversed phase (RP), normal phase (NP) and ion exchanger. Reversed Phase Normal Phase Ion Exchanger Typical extraction mechanisms Extraction of hydrophobic or polar organic analytes from aqueous matrix Extraction of polar analytes from non-polar organic solvents Extraction of charged analytes from aqueous or non-polar organic samples Types of retention mechanisms: Nonpolar interactions silica-based: C18 ec, C18, C18 Hydra, C8, polymer-based: HR-X, HR-P, Easy, PS-RP interactions: hydrophobic sample: mostly aqueous elution: solvents with lower polarity (compared to water) MeOH, CH Cl, CHCl 3, hexane Polar interactions silica-based: SiOH, CN, NH, OH (diol), C 6, other: Alox, Florisil, interactions: hydrogen bonds, dipole-dipole and π-π interactions sample: mostly organic elution: polar solvents (compared to sample solvent) (nonprotic) ethers, ketones (MTBE, THF, acetone, ) CH Cl, CHCl 3, Cation exchangers silica-based: SA (SCX), PCA (WCX), PSA, polymer-based: HR-XC, HR-XCW, PS-H +, interaction: between charged analytes and functional group of cation exchanger sample: aqueous (ph3-5) elution: acidic: ph (e. g. HCl, or 0% AcOH in MeOH/acetonitrile) basic: ph8-9 (e. g. 5% NH 3 in MeOH/acetonitrile) solvents or buffers with higher ionic strength and counter ions with high selectivity Anion exchangers silica-based: SB (SAX), NH, DMA, polymer-based: HR-XA, HR-XAW, PS-OH -, interaction: between charged analytes and functional group of anion exchanger sample: aqueous (ph 8-9) elution: basic: ph10 (e. g. 0% NH 3 in MeOH/acetonitrile) acidic: ph4-5 (e. g. HCl, or 5% AcOH in MeOH/acetonitrile) solvents or buffers with higher ionic strength and counter ions with high selectivity Suggested loading capacity Cartridge size Condition Equilibrate Load Wash Elute 1ml 1 ml 1 ml 1 ml 1 ml 1 ml 3ml ml ml ml ml ml 6 ml 3 ml 3 ml 5 ml 4 ml 4 ml Who can use SPE? - Any analytical laboratory can employ SPE and its benefits - Environmental agencies - Clinical laboratories - Food control laboratories and food producers Watchers bond volume capacity 1ml 3ml 6ml 15ml Watcher bond Polypropylene Traditional SPE Provides a variety of adsorbents Volume : 1ml, 3ml, 6ml, 15ml Ads Weight : 100mg, 00mg, 500mg, 1g, g 5
6 Solid Phase Extraction SPE method development kits If you do not know the behaviour of your samples or analyte in solid phase extraction, or if you want to optimise an existing extraction procedure, we recommend Watchers method development kits. Consisting of a variety of different SPE phases and column formats they are the ideal tool for approaching any analytical task systematically. Ordering information Designation Contents of the kit Investigating the best separation mechanism for a clean-up procedure Watchers bond HR-Xpert columns with 3ml / 60mg each: 10columns with HR-X, development kit I 5columns each with HR-XC, HR-XA, HR-XCW, HR-XAW Watchers bond HR-Xpert columns with 3ml / 00mg each: 10columns with HR-X, development kit II 5columns each with HR-XC, HR-XA, HR-XCW, HR-XAW Watchers bond columns with 3ml / 00mg each: 10columns with HR-X, standard development kit 5columns each with 3ml / 500mg: C18, C18 ec, C8, C 6, NH, DMA, OH, CN, SiOH, SA (SCX), SB (SAX) Selecting the optimum RP phase for a clean-up procedure Watchers bond RP development kit I 10columns each with 6ml / 1000mg: C18, C18 ec, C8, C4 Watchers bond RP development kit II 10columns each with 6ml / 1000mg: C18, C18 ec, C8, C4, HR-P, HR-X Watchers bond RP development kit III 10columns each with 6ml / 1000mg: C18, C18 ec, C18 Hydra, C8 Watchers bond RP development kit IV 10columns each with 6ml / 1000mg: C18, C18 ec, C18 Hydra, C8, HR-P, HR-X Selecting the optimum polar phase for a clean-up procedure Watchers bond polar development kit I 10columns each with 6ml / 1000mg: SiOH, Florisil, NH, CN, OH Watchers bond polar development kit II 10columns each with 6ml / 1000mg: SiOH, Florisil, NH, CN, OH Customized Selection phase for clean-uo procedure Watchers bond polar development kit I 10columns each with 6ml / 1000mg: C18, C8, C4, SiOH, Florisil, NH Costomised SPE Column In order handle the increasing requests for custom-made Watchers bond column Column bed: adsorbent:. Weight:.mg Column Volumn: Volumn:. (polyethlene) Required Quantity of column: Costomised SPE Column Comparison of recovery phases for phenol analysis Comparison of different phases for phenol analysis Compounds investigated: phenol,, 4-dinitrophenol, pentachlorophenol Column types: Watcher bond C18 / 6ml / 000mg Watcher bond C 6 H 11 ec / 6ml / 000mg Column conditioning: 10ml acetone, 10ml methanol, and 10ml dist. Water (ph) Sample application: Aspirate the sample through the column Elution: 10ml methanol 6
7 Watchers bond Ordering information Volume 1ml 3ml 6ml 15ml Sorbent Mass 100mg 00mg 500mg 500mg 1g g g Q ty per Box Reversed Phase Watchers bond Easy SR SR30-50 SR SR SR SR60-30 SR150-0 Watchers bond C18ec SC SC SC SC SC SC SC Watchers bond C8 SC SC SC SC SC SC SC Watchers bond C 6 SR SR SR SR SR SR SR Normal phase Watchers bond SiOH SS SS SS SS SS SS SS Watchers bond Florisil SF SF SF SF SF SF SF Watchers bond NH SN SN SN SN SN SN SN Watchers bond CN SN SN SN SN SN SN SN Watchers bond Alox A SN SN SN SN SN SN SN Watchers bond Alox N SN SN SN SN SN SN SN Watchers bond Alox B SN SN SN SN SN SN SN Ion Exchangers Watchers bond SB SI SI SI SI SI SI SI Watchers bond SA SI SI30-50 SI SI SI SI60-30 SI150-0 Watchers bond HR-XC SI SI SI SI SI SI SI Watchers bond HR-XA SI SI SI SI SI SI SI Watchers bond & CHROMABOND QuEChERS Base material silica, pore size 60Å, particle size 45μm, specific surface 500m /g ph stability -8 Primary and Secondary Amine functions (PSA), 5%C removes polar compounds (e.g., organic acids, pigments, sugars) from matrices like fruit or vegetables Similar phases: Supelclean PSA, Bond Elut PSA Recommended application: special SPE phase for quick and cheap determination of pesticides in strongly matrix-contaminated samples by GC-MS and LC-MS QuEChERS method = Quick Easy Cheap Effective Rugged Safe Sample property Low fat content Moderate content of Higher content of chlorophyll Higher fat content chlorophyll and carotinoids and carotinoids (e.g., apples, strawberries) (e.g., carrots, lettuce) (e.g., bell peppers, spinach) (e.g., avocado) CHROMABOND QuEChERS extraction mixes Citrate or acetate extraction Citrate or acetate extraction Citrate extraction Citrate extraction Mix I or Mix II Mix I or Mix II Mix II Mix II CHROMABOND QuEChERS clean-up mixes Diamino clean-up Diamino/Carbon clean-up Diamino/Carbon clean-up Diamino/C18 ec clean-up (higher Carbon content) Mix III Mix IV Mix V Mix VI 7
8 Solid Phase Extraction Ordering information Volume Description Composition Composition REF Pack CHROMABOND QuEChERS extraction buffer mixes 15ml Mix I citrate extraction mix 4g MgSO 4, 1g NaCl, 0.5g Na H citrate H O, 1g Na 4 citrate H O 15ml Mix II acetate extraction mix 6g MgSO 4, 1.5g Na acetate Easy Oasis 50 HR-P CHROMABOND QuEChERS clean-up mixes 15ml Mix III Diamino clean-up mix 0.15g CHROMABOND Diamino with 0.9g MgSO 4 15ml Mix IV Diamino/Carbon 0.15 g CHROMABOND Diamino with 0.9g clean-up mix MgSO 4 and 15mg Carbon 15ml Mix V Diamino/Carbon 0.15g CHROMABOND Diamino with 0.9g clean-up mix MgSO 4 and 45mg Carbon 15ml Mix VI Diamino/C18 ec 0.15g CHROMABOND Diamino with 0.9g clean-up mix MgSO 4 and 150mg C18 ec CHROMABOND Diamino polypropylene columns 3ml adsorbent weight 00mg ml adsorbent weight 500mg CHROMABOND Diamino adsorbent 15ml centrifuge tubes with screw cap (ml or 50mL centrifuge tubes on request) g g 주소서울특별시서초구언남1길 16 삼익빌딩 4층 Tel Fax Add 4F Samik B/D, Eonnam 1gil16, Seocho-gu, Seoul, Korea Tel Fax
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