Polymer Coated Titania for Analytical and Preparative Reversed-Phase Chromatography
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1 Polymer Coated tania for Analytical and Preparative Reversed-Phase Chromatography EAS 200 BIGWE YA 1, CLAYT V. MCEFF 1, JCHE WIKLER 2 1 ZirChrom Separations, Inc., 617 Pierce Street, Anoka, M Sachtleben, Duisburg, Germany STABLE-1 1
2 utline Properties of tania RP Phase Stationary Phase Chromatographic Data Selectivity Comparison of Sachtopore-RP, Silica C18, ZirChrom-CARB, and ZirChrom-PBD Chemical and Thermal Stability Testing Effect of Lewis Base Mobile Phase Additive on Elution of Basic Compounds Conclusion - The Sachtopore -RP shows very similar selectivity to ZirChrom -PBD and has excellent stability from ph 1-12 and up to 100 o C. 2
3 Surface Chemistry of tania- Based Supports for HPLC H H H 2 H H 2 H 2 Weak Brönsted Acid: Weak Brönsted Base: Strong Lewis Acid: H + H + H2 + H + P - + SCX P H mode 3
4 Polyethylene Coated tania [ CH 2 -CH ] 2 n Polyethylene H H H 2 H H 2 H 2
5 22 on-electrolyte Solutes onpolar Benzene Toluene Ethylbenzene p-xylene Propylbenzene Butylbenzene Cl Polar Br Cl Bromobenzene p-dichlorobenzene Anisole Methylbenzoate aphathalene Acetonphenone 2 C 2 2 Cl Benzonitrile itrobenzene p-itrotoluene p-itrobenzyl Chloride Benzophenone Cl HB Donor H H H H H H Benzylalcohol 3-Phenyl Propanol -Benzyl Formamide Phenol p-chlorophenol
6 Selectivity Comparison ZirChrom-PBD ZirChrom-CARB Silica C18 Sachtopore -RP benzyl formamide benzyl alcohol phenol 3-phenyl propanol p-chlorophenol acetophenone benzonitrile nitrobenzene methylbenzoate anisole p-chlorotoluene p-nitrobenzyl chloride benzophenone bromobenzene naphthalene p-xylene p-dichlorobenzene benzene toluene ethyl benzene propyl benzene butyl benzene Solute LC Conditions: Mobile phase, 0/60 AC/Water; Flow rate, 1.0 ml/min.; Temperature, 30 o C; Injection volume, ul; Detection at 2nm. 6 log (k' Solute/k' benzene)
7 κ-κ Comparison k' (PBD, CARB, C18) Sachtopore -RP vs Silica-C18 Sachtopore -RP vs ZirChrom-CARB Sachtopore -RP vs ZirChrom-PBD R 2 = R 2 = R 2 = k' (Schtopore RP) LC Conditions: Mobile phase, 0/60 AC/Water; Flow rate, 1.0 ml/min.; Temperature, 30 o C; Injection volume, ul; Detection at 2nm. Reference: Melander, W.; Stoveken, J.; Horvath, C. J. Chromatogr. 1980, 199,
8 Chemical Stability ph 1.0 ph toluene toluene.0 methyl benzoate.0 Methyl benzoate k' k' 2.0 anisole 2.0 anisole Column Volumes Benzonitrile Column Volumes Benzonitrile Exposure and Evaluation Conditions: Mobile phase, 1/8 AC/0.1M itric acid, ph 1.0, or 0.01M Tetramethylammoniumhydroxide, ph 12.0; Flow rate, 1.0 ml/min.; Temperature, 30 o C; Injection volume, μl; Detection at 2 nm; Column, 0 mm x.6 mm i.d., Sachtopore -RP (part# TI01-06). 8
9 Temperature Stability at 100 o C 3.0 toluene 2.0 k' 1.0 methyl benzoate anisole Benzonitrile Column Volumes Exposure and Evaluation Conditions: Mobile phase, 1/8 AC/water; Flow rate, 1.0 ml/min.; Temperature, 100 o C with Metalox heater; Injection volume, ml; Detection at 2 nm; Column, 0 mm x.6 mm i.d., Sachtopore -RP (part# TI01-06). 9
10 Effect of Lewis Base Additive on Separation of Basic Drugs A 2 B C min LC Conditions: Mobile phase: 30/70 AC/20 mm buffer (ph=7). (A) ammonium acetate, (B) ammonium fluoride, (C) ammonium phosphate. flow rate: 1ml/min, temperature: 0 o C. Wavelength: 2 nm. Column, 0 mm x.6 mm i.d., Sachtopore -RP (part# TI01-06). Solutes: (1) lidocaine, (2) quinidine, (3) tryptamine, () amitriptyline, and () nortriptyline. 10
11 Effect of Ionic Strength on Separation of Basic Drugs (A) 10 mm (B) 1 mm (C) 20 mm min LC Conditions: Mobile phase: 30/70 AC/phosphate buffer (ph=7). (A) 10 mm, (B) 1 mm, (C) 1 mm. flow rate: 1ml/min, temperature: 0 o C. Wavelength: 2 nm. Column, 0 mm x.6 mm i.d., Sachtopore -RP (part# TI01-06). Solutes: (1) lidocaine, (2) quinidine, (3) tryptamine, () amitriptyline, and () nortriptyline. 11
12 Antihistimines Separation at ph 10,0 mau 230 nm 0,0 3 H H CH C H H 3 C 30, : maleic acid : Diphenylpyraline CH 3 CH3 Cl 20,0 2: Carbinoxamine CH 3 CH 3 Cl CH3 10,0 H 3 C 3: Chlorphenamine : Phenyltoloxamine 0,0 -,0 0,0 2,0,0 6,0 8,0 min 10,0 12,0 1,0 LC Conditions: Mobile phase, 7% (0 mm H 3 P + mm KH 2 P ), 26% AC, ph 10; Flow rate, 1.0 ml/min.; Temperature, Ambient; Injection volume, 20 μl; Detection at 220 nm; Column, Sachtopore -RP (300 Å, 3 µm, 10x mm). 12
13 Separation of Basic Cardiac Drugs at ph 10 33,0 2,0 20,0 mau H 3 C CH 3 H 3 C 1: Trapidil H H 3 C C H 3 H 3 C CH3 2 H 3 C CH 3 H H C H 3 CH 3 2: Metoprolol I I C H 3 CH 3 H H 230 nm H H : Dipyridamole 3: ifedipine : Amiodarone H 3 C C H 3 1,0 CH 3 7: Etafenone 6 H 3 C C CH 3 10,0 H 3 C H 3 C CH 3 CH 3 7 6: Verapamil,0 min -1,0 0,0 2,0,0 6,0 8,0 10,0 12,0 1,0 LC Conditions: Mobile phase, 70% (10 mm Borax + 10 mm Soda), 30% AC, ph 10; Flow rate, 1.0 ml/min.; Temperature, Ambient; Injection volume, 20 μl; Detection at 230 nm; Column, Sachtopore -RP (300 Å, 3 µm, 10x mm). 13
14 Semi-Prep Separation of Pentifylline (vasodilator) 1800 SLB16 #69 A1 UV_VIS-1 mau 280 nm Particle Sizes: 3,,10,20,0, micron 1 mm Pores sizes: 60, 100, 300, 00, 1000, 2000 Angstroms min 0,0 1,0 2,0 3,0,0,0 6,0 7,0 8,0 9,0 10,0 LC Conditions: Mobile phase, (+ 10 mm a 2 B mm H 3 B 3 ), ph 8.8; Flow rate, 1.0 ml/min.; Temperature, Ambient; Injection volume, 20 μl; Detection at 2 nm; Column, Sachtopore -RP (300 Å, 3 µm, 10 x mm). 1
15 Conclusions The Sachtopore -RP shows similar selectivity to ZirChrom -PBD (DS-like for neutrals). The Sachtopore -RP has excellent stability from ph 1-12 and up to 100 o C. The type of Lewis base buffer has a profound effect on selectivity for Sachtopore -RP. Basic (amine) analytes generally undergo RP/CEX mixed-mode retention mechanism on Sachtopore -RP. 1
16 For More Information, Visit Us at Booth STABLE-1 For more information and web access to the free Buffer Wizard: 16
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