BETASIL Columns TG Å high purity silica

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1 BETASIL Columns TG0-0 00Å high purity silica

2 Technical Guide BETASIL Columns Introduction 00Å high purity silica High surface area and phase loading Designed for small molecules Type B silica gives good results with acids or bases High phase loading means mobile phases require more organic solvent (popular for mass spec) All reversed phases are end-capped Dense bonding results in excellent reproducibility and stability Excellent performance with acids, bases and neutral compounds BETASIL Columns Specifications Phase Particle size Carbon Load Pore Size Endcapping Silica type BETASIL C8, and 0µm 0% 00Å Yes High purity, BETASIL C8, and 0µm % 00Å Yes High purity, BETASIL C and µm % 00Å Yes High purity, BETASIL C and µm % 00Å Yes High purity, BETASIL Phenyl µm % 00Å Yes High purity, BETASIL Phenyl/Hexyl and µm % 00Å Yes High purity, BETASIL Cyano µm % 00Å Yes High purity, BETASIL Diol 00 µm % 00Å Yes High purity, BETASIL Silica 00 µm N/A 00Å N/A High purity, BETASIL Chromatographic Characterization The BETASIL C8 phase is a high carbon load packing that offers up to twice the retention of other C8 packings with lower percent carbon loading. Figure shows the increased retention of the BETASIL C8 column when compared to other Thermo Hypersil-Keystone columns. The increased retention associated with the BETASIL column is directly attributed to the increased surface area of theparticles, and higher carbon loading than both Hypersil BDS C8 and BetaBasic 8 columns. Where longer retention is required or there is a requirement to use a higher percentage of organic solvent in themobile phase, e.g. to increase LC-MS sensitivity, the BETASIL C8 packing offers an excellent choice. Figure. Increased Hydrophobic Retention of Neutral Molecules BETASIL C8 0%C MIN Columns: µm, 0x.mm Eluent: % ACN / % H Flow:. ml/min Detector: Uracil. Butyl Benzene. Benzene. Pentyl Benzene. Ethyl Benzene. Hexyl Benzene. Propyl Benzene 8. Heptyl Benzene BetaBasic 8 %C 8 Hypersil BDS C8 %C MIN 0 0 MIN 8

3 Stability at Low ph All surface bonded chemistries belonging to the BETASIL family are highly stable under chemical attack. Hydrolysis of the silane ligand can occur under acidic conditions, resulting in a loss of retention and/or selectivity. The effect becomes more severe as ligand chain length is reduced, as is observed for the BETASIL C phase. Wettable stationary phases such as the BETASIL Cyano packing can also show slightly less resistance to chemical attack, and consequently are recommended only for use between ph and 8. In general, BETASIL packings show superior stability to many other packings. Figure shows BETASIL phases subjected to,000 column volumes of aggressive mobile phase at ph.8 and 0 C. The BETASIL C8 phase exhibits virtually no change in retention for the analysis of a sensitive drug mixture. Base Deactivated for Better Peak Shape with Basic Compounds All BETASIL packings are base deactivated and show excellent peak shape for basic compounds. Figure shows the analysis of a procainamide test mixture, a mixture that would typically have given rise to broad tailing peaks for the more basic compounds on traditional C8 silicas prepared on a Type A silica. In terms of silanol activity, the BETASIL packing is similar to other Type B silica-based phases such as BetaBasic and BetaMax Neutral phases. Phase Collapse Phenomenon Mobile phases that contain a high proportion of water often are employed to retain highly polar compounds when using RP-HPLC. Many C8 columns will show a Figure. BETASIL Phase Stability At Low ph reversible loss of retention when exposed to highly aqueous mobile phase. The rate and degree of retention loss can vary greatly among different columns. A common explanation for this Figure. Base Deactivation of BETASIL C. Uracil. Procainamide. N-Acetyl Procainamide. Caffeine. N-Propionyl Procainamide MIN BETASIL C, µm, 0x.mm Eluent: 0% ACN / 90% 0.0M Phosphate buffer, ph. with H P Flow:. ml/min Detector: 0.% TFA gradients at 0 C, ph.8 retention loss is that the hydrophobic alkyl chains of the stationary phase are not wettable and appear to fold down on the silica surface to avoid a highly aqueous, hydrophilic mobile phase (Figure ). Figure. Phase Collapse

4 Technical Guide BETASIL Columns Regeneration After Phase Collapse In this folded or collapsed state, the alkyl chains are much less able to interact with solutes, resulting in a loss of retention. We use the term chain folding to refer to this reversible loss of retention induced by highly aqueous mobile phases, although the exact mechanism of reversible retention loss is not proven. The BETASIL phase is a high carbon load, high density C8 and consequently shows the characteristics associated with chain folding, i.e. loss in retention. However, the situation is completely reversible and the column can be regenerated simply by flushing with 0% ACN / 0% H for 0 minutes (Figure ). The phenomena of chain folding can be almost completely overcome by including trace amounts of organic solvent in the mobile phase. Figure shows how just 0.% n-propanol has been used to retard the loss in retention associated with chain collapse. Further information on chain collapse can be found in Technical Bulletin TB99-0. Choices in Stationary Phase Chemistries BETASIL columns are available in a wide range of reversed phase packings, including C8, C8, C, Phenyl, Cyano, and Phenyl/Hexyl. The BETASIL Phenyl/Hexyl phase provides a combination of straightchain C groups and phenyl groups, resulting in a mixed-mode separation. The C chain exhibits classical reversed phase retention, while the phenyl ring provides special selectivity for polar groups (Fig. ). BETASIL columns also offer choices for normal phase chromatography, including silica and diol phases in different pore sizes from 0Å to 00Å, and cyano. Diol phases provide higher polar selectivity than cyano with less water sensitivity than bare silica. Figure. High Carbon Load C8 in 00% Aqueous Mobile Phase,,,. Tartaric acid. Malonic acid. Fumaric acid. Succinic acid bet800 bet800 bet MIN 0 8 MIN 0 8 MIN Partially Folded Completely Folded after > hours use Regenerated (0% ACN / 0% H for 0 minutes) BETASIL C8, 0x.mm Eluent: 0.0M KH P + 0.0M H P Flow:.0 ml/min Detector: 0

5 Figure. Trace rganics in Mobile Phase Retard Chain Folding Figure. Polyphenols. Tartaric acid. Malonic acid. Fumaric acid. Succinic acid. Pyrogallol. Hydroquinone. Resorcinol. Phenol Initial bet80 After 8 hours H0-00 bet800 0 MIN BETASIL C8, 0x.mm Eluent: 0.0M KH P + 0.0M H P + 0.% n-propanol Flow:.0 ml/min Detector: MIN BETASIL Phenyl/Hexyl, µm, 0x.mm Eluent: mm Acetic Acid Flow:.0 ml/min Detector: 80 Temp.: C rganic Acids Benzodiazepines. Ascorbic Acid. Acetaminophen. Benzoic Acid. Salicylic Acid. Nitrazepam. xazepam. Temazepam. Nordiazepam. Diazepam. Medazepam H MIN BETASIL Phenyl/Hexyl, µm, 0x.mm Eluent: 80% 0mM KH P, ph / 0% MeH Flow:.0 ml/min Detector: Temp.: C BETASIL Phenyl/Hexyl, µm, 0x.mm Gradient: A: H B: MeH Time %B 0 8 Flow:.0 ml/min Detector: Temp.: C H MIN

6 Technical Guide BETASIL Columns Carbamate/Urea Pesticides & Herbicides. Tebuthiuron. Carbofuran. Carbaryl. Diuron. Barban Procainamides. Uracil. Procainamide. N-Acetyl Procainamide. Caffeine. Propionyl Procainamide H MIN BETASIL Phenyl/Hexyl, µm, 0x.mm Gradient: A: H B: ACN Time %B 0 8 Flow:. ml/min Detector: Temp.: C 0 8 MIN BETASIL C, µm, 0x.mm Eluent: 0% ACN / 90% 0.0M KH P, ph. with H P Flow:. ml/min Detector: Cyclosporin Assay 00 ng / ml standard. Cyclosporin. Internal Standard Real 8 ng / ml c-std c-pat 0 MIN 0 MIN BETASIL C, µm, 0x.mm Eluent: % ACN / % 0.0M KH P, ph Flow:. ml/min Detector:

7 Common Analgesic. Acetaminophen. Caffeine. Acetylsalicylic Acid. Benzoic Acid Water-Soluble Vitamins. Pyridoxine. Thiamine. Niacinamide. Cyanocobalamin. Riboflavin. Dimethoate. Methyl Parathion. Malathion. Fenitrothion. Ethyl Parathion (DNTP). Diazinon. EPN Pesticides MIN BETASIL C8 PINEER TM, µm, 0xmm Eluent: A: 0.0M Ammonium Acetate, ph B: 0% ACN% / 0% MeH 8: A:B Flow:. ml/min Detector: vit MIN BETASIL C8, µm, 0x.mm Gradient: A: 0.0M K HP, ph. B: ACN % B to 0% B in min Flow:.0 ml/min Detector: beta MIN BETASIL C8, µm, 0x.mm Gradient: 0% ACN / 0% H, step to 0% ACN / 0% H at min. Flow:.0 ml/min Detector: Steroids by Normal Phase. Progesterone. 0α-Hydroxy--pregnene--one. α-hydroxyprogesterone. Deoxycorticosterone. α-hydroxyprogesterone Acetate. α-hydroxyprogesterone. α-ketoprogesterone 8. Corticosterone 9. Cortisone Acetate 0. Hydrocortisone. Cortisone,, , MIN MIN 0 MIN BETASIL Diol 0, µm, 0x.mm Eluent: 8% Isooctane /.% EtH / 0.% H Flow:. ml/min Detector: BETASIL Diol 00, µm, 0x.mm Eluent: 8% Isooctane /.% EtH / 0.% H Flow:. ml/min Detector: BETASIL Diol 00, µm, 0x.mm Eluent: 8% Isooctane /.% EtH / 0.% H Flow:. ml/min Detector:

8 Innovators in life and laboratory sciences, Thermo Electron Corporation provides advanced analytical technologies, scientific instrumentation, laboratory informatics solutions, and laboratory consumables to help scientists and clinicians to discover new drugs, improve manufacturing processes, and diagnose illness and disease. Unparalleled in our capabilities, we can help you every step of the way from sample preparation and sample analysis through interpretation of results. Thermo Electron Corporation has direct subsidiary offices in North America, Europe and Japan. To complement these direct subsidiaries, we maintain a network of representative organizations throughout the world. Use this reference list or visit our web site to locate the representative nearest you. France Tel. () Germany Tel. (9) United Kingdom Tel. () United States Tel. (0) For more information on our products and services, please visit our website at: Technical information contained in this publication is for reference purposes only and is subject to change without notice. Every effort has been made to supply complete and accurate information; however, Thermo Electron assumes no responsibility and will not be liable for any errors, omissions, damage, or loss that might result from any use of the information contained therein (even if this information is properly followed and problems still arise). Reference to specifications supersede all previous information and are subject to change without notice. ADVANCE, BetaBasic, BetaMax, BETASIL, BioBasic, DASH, DELTABND, Duet, Fluophase, Hyperbond, Hypercarb, KAPPA, HT PCKET, HyperGEL, HyperREZ, HyperSEP, Hypersil, HyPURITY, HyPURITY AQUASTAR, Javelin, KEYSTNE, MultiSEP, PINEER, PRISM, Retain, SLIPFREE, UNIGUARD, UNIPHASE, and Verify are trademarks of Thermo Electron Corporation. AquaSil TM Siliconizing Fluid for treating glass surfaces is sold by Pierce Chemical Co., Rockford, IL. All other trademarks are the property of their respective owners Thermo Electron, Printed in USA 0/0.

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