Polymer Coated tania for Analytical and Preparative Reversed-Phase Chromatography Pittcon 2005 CLAYT V. MCEFF 1, BIGWE YA 1, JCHE WIKLER 2 1 ZirChrom Separations, Inc., 617 Pierce Street, Anoka, M 55303 2 Sachtleben, Duisburg, Germany 1-866-STABLE-1 www.zirchrom.com
utline Properties of tania RP Phase Stationary Phase Chromatographic Data Selectivity Comparison of Sachtopore-RP, Silica C18, ZirChrom-CARB, and ZirChrom-PBD Effect of Lewis Base Mobile Phase Additive on Elution of Basic Compounds Chemical and Thermal Stability Testing 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.
Element Electronic Structures Silicon (element 14; 2.3 g/cc; Si 4+ ionic radius 0.400 Å) e3s 2 3p 2 tanium (element 22; 4.5 g/cc; 4+ ionic radius 0.605 Å) Ar3d 2 4s 2 Zirconium (element 40; 6.5 g/cc; Zr 4+ ionic radius 0.720 Å) Kr4d 2 5s 2 All have four valence electrons so some chemistry is similar, but presence of d orbitals and very electropositive nature allow and Zr (metals) to form strong electron donor-acceptor complexes (coordination chemistry).
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 P + H + P - H SCX mode
tania Crystal Forms Crystallographic data and pi values of sorbents Anatase (tetragonal), 300 o C Rutile (tetragonal), 600 o C Brookite (orthorhombic), 850 o C. Si 2 2 Al 2 3 Zr 2 Ionisation potential (V) 45.14 (Si 4+ ) 43.27 ( 4+ ) 28.45 (Al 3+ ) 34.34 (Zr 4+ ) Coordination number 4 6 6 7 Ion radius (A) 0.41 (Si 4+ ) 0.68 ( 4+ ) 0.5 (Al 3+ ) 0.8 (Zr 4+ ) Ionisation potential/coordination number (V) 11.29 7.33 4.74 4.83 pi of sorbent (ph) 2.2 6.6 8.1 6 Porosities of test columns (%) 73 74 79 69 Anatase Rutile
Polyethylene Coated tania [ CH 2 -CH ] 2 n Polyethylene H H H 2 H H 2 H 2
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
Selectivity Comparison 2.000 1.500 ZirChrom-PBD ZirChrom-CARB Silica C18 Sachtopore -RP 1.000 0.500 0.000-0.500-1.000 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 -1.500 benzene toluene ethyl benzene propyl benzene butyl benzene Solute LC Conditions: Mobile phase, 40/60 AC/Water; Flow rate, 1.0 ml/min.; Temperature, 30 o C; Injection volume, 5ul; Detection at 254nm. log (k' Solute/k' benzene)
κ-κ Comparison 45.0 k' (PBD, CARB, C18) 40.0 35.0 30.0 25.0 20.0 15.0 Sachtopore -RP vs Silica-C18 Sachtopore -RP vs ZirChrom-CARB Sachtopore -RP vs ZirChrom-PBD R 2 = 0.911 R 2 = 0.036 10.0 5.0 R 2 = 0.992 0.0 0.00 1.00 2.00 3.00 4.00 5.00 k' (Schtopore RP) LC Conditions: Mobile phase, 40/60 AC/Water; Flow rate, 1.0 ml/min.; Temperature, 30 o C; Injection volume, 5ul; Detection at 254nm. Reference: Melander, W.; Stoveken, J.; Horvath, C. J. Chromatogr. 1980, 199, 35-56.
Chemical Stability ph 1.0 ph 12.0 6.0 6.0 toluene toluene k' 4.0 methyl benzoate k' 4.0 Methyl benzoate 2.0 anisole 2.0 anisole Benzonitrile 0.0 0 1000 2000 3000 4000 5000 Column Volumes 0.0 Benzonitrile 0 1000 2000 3000 4000 5000 Column Volumes Exposure and Evaluation Conditions: Mobile phase, 15/85 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, 5 µl; Detection at 254 nm; Column, 50 mm x 4.6 mm i.d. Sachtopore -RP.
Temperature Stability at 100 o C 3.0 toluene 2.0 k' 1.0 methyl benzoate anisole Benzonitrile 0.0 0 1000 2000 3000 4000 5000 6000 Column Volumes Exposure and Evaluation Conditions: Mobile phase, 15/85 AC/water; Flow rate, 1.0 ml/min.; Temperature, 100 o C with Metalox heater; Injection volume, 5 ml; Detection at 254 nm; Column, 50 mm x 4.6 mm i.d., Sachtopore -RP.
Effect of Lewis Base Additive on Separation of Basic Drugs 2 1 3 4 5 A 2 1 3 4 5 B 1 2+3 4 C 5 0 2 4 6 8 10 12 14 16 18 min Chromatographic Conditions: Column 50X4.6 mm Sachtopore -RP, Mobile phase: 30/70 AC/20 mm buffer (ph=7). (A) ammonium acetate, (B) ammonium fluoride, (C) ammonium phosphate. flow rate: 1ml/min, temperature: 40 o C. Wavelength: 254 nm. Solutes: (1) lidocaine, (2) quinidine, (3) tryptamine, (4) amitriptyline, and (5) nortriptyline.
Effect of Ionic Strength on Separation of Basic Drugs (A) 10 mm 1 2 3 4 5 2 1 3 4 5 (B) 15 mm 1 2+3 4 5 (C) 20 mm 0 2.5 5 7.5 10 12.5 15 17.5 20 22.5 min Chromatographic Conditions: Column 50X4.6 mm Sachtopore -RP, Mobile phase: 30/70 AC/phosphate buffer (ph=7). (A) 10 mm, (B) 15 mm, (C) 15 mm. flow rate: 1ml/min, temperature: 40 o C. Wavelength: 254 nm. Solutes: (1) lidocaine, (2) quinidine, (3) tryptamine, (4) amitriptyline, and (5) nortriptyline.
Antihistimines Separation at ph 10 45,0 mau 230 nm 40,0 H CH 3 5 H CH H 3 C 30,0 1 2 4 1: maleic acid 4: Diphenylpyraline Cl 20,0 2: Carbinoxamine Cl CH3 H 3 C 5: Phenyltoloxamine 10,0 3: Chlorphenamine 0,0-5,0 0,0 2,0 4,0 6,0 8,0 min 10,0 12,0 15,0 LC Cconditions: Mobile phase, 74% (50 mm H 3 P 4 + 5 mm KH 2 P 4 ), 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, 150x4 mm).
Separation of Basic Cardiac Drugs at ph 10 33,0 mau 230 nm H H 3 C H C H 3 H 25,0 1 2 3 H 3 C 1: Trapidil H H 3 C C H 3 H 3 C H 3 C 2: Metoprolol I H H H 5: Dipyridamole C H 3 20,0 2 I 3: ifedipine 4: Amiodarone H 3 C C H 3 15,0 5 7: Etafenone 6 H 3 C C 10,0 4 H 3 C H 3 C C H 3 C H 3 6: Verapamil 7 5,0-1,0 0,0 2,0 4,0 6,0 8,0 10,0 12,0 15,0 LC Cconditions: 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, 150x4 mm). min
Semi-Prep Separation of Pentifylline (vasodilator) 1800 SLB16 #69 A15 UV_VIS-1 mau 280 nm Particle Sizes: 3,5,10,20,40,80 100 micron 1 mm 1500 1250 1000 750 Pores sizes: 60, 100, 300, 500, 1000, 2000 Angstroms 500 250 0 min -200 0,0 1,0 2,0 3,0 4,0 5,0 6,0 7,0 8,0 9,0 10,0 LC Cconditions: Mobile phase, (+ 10 mm a 2 B 4 7 + 1 mm H 3 B 3 ), ph 8.8; Flow rate, 1.0 ml/min.; Temperature, Ambient; Injection volume, 20 µl; Detection at 254 nm; Column, Sachtopore -RP (300 Å, 3 µm, 150 x 4 mm).
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.
Acknowledgments Dr. Jochen Winkler Sachtleben Dr. Bingwen Yan - ZirChrom Visit Us at Booth 2801 1-866-STABLE-1 www.zirchrom.com For more information and web access to the free Buffer Wizard: www.zirchrom.com