U.S. Patent o. 7, 5, 7 Setting the Standard for ph Method Development ugged reversed phase HPLC columns that offer extended lifetime at extreme ph conditions and excellent stability for reproducible, high efficiency separations. Take full advantage of high and low ph conditions (ph -) to manipulate selectivity Expect longer column lifetime with patented TWI-X technology For analytical and preparative separations of basic and acidic compounds If Gei analytical columns do not provide at least an equivalent separation as compared to a competing column of similar particle size, similar phase and dimensions, send in your comparative data within 5 days and keep the Gei column for FEE. Phase X C8 C-Phenyl C8 Description The most rugged Gei column, offering 5 times the durability of previous generation hybrid columns A low bleed phenyl phase. For UV and MS detection, which offers an aromatic selectivity complementary to C8 phases Selectivity, high structural integrity and increased loadability for preparative and purification applications in pre-packed columns and bulk media USP Classification L L L APPVED TWI (Two-In-ne) Technology During the final stage of silica manufacturing a unique silicaorganic layer is grafted to create a completely new composite particle. Since the internal base silica is unaltered by this manufacturing process, the particle retains its mechanical strength and rigidity along with excellent efficiency, while the silica-organic shell protects the particle from chemical attack. Second-Generation TWI-X Technology TWI-X technology uses an improved patented organo-silica grafting process which incorporates highly stabilizing ethane cross-linking. These organic groups are evenly incorporated into the grafted layers on the silica surface while maintaining a pure silica core. This not only provides resistance to high ph attack, but also maintains the high efficiency and mechanical strength of a silica particle. HPLC GEMII Pore Si Si Si S Si Si Si Si C Si Si Si Si Si Si Si Si C C Si Si Si Si Si Si Si Si Gei is a registered trademark of Phenomenex, Inc. TWI-X is a trademark of Phenomenex, Inc. Phenomenex 5
U.S. Patent o. 7, 5, 7 Gei-X C8 Control selectivity of ionizable compounds for optimized methods Consistent performance in both volatile and non-volatile buffers High sample loading capacity for metabolite identification and preparative purification ph stable - for durability Gei-X C8 USP: L LC/MS Certified Hydrophobicity H-Bonding Aromatic Selectivity Silanol Activity (ph.5) Low see p. 7 for testing probes. ph Stability.0-.0 Particle Size, 5, and 0 μm Phase C8 Application Small Molecules, Basic Compounds Strength Most durable Hybrid Particle High APPVED GEMII HPLC Gei-X Tested for Extreme Durability in Changing Mobile Phase ph Column Efficiencies Maintained in High ph Testing for 0 Cycles % of Initial Efficiencies etention in Minutes Material Characteristics Packing Material Particle Shape/Size (µm) Pore Size (Å) Surface Area (m /g) Carbon Load % End Capping Gei-X C8 Spherical, 5,0 0 75 TMS 0 00 90 80 70 0 50 0 5 0 5 0 5 Diphenhydrae Tetracaine Stable Efficiencies After Exposure to 0,000 Column Volumes of High and Low ph Buffers Cycles of Exposure etention Times of Four Probes Maintained in eutral ph Testing for 0 Cycles etention Times Maintained After Exposure to 0,000 Column Volumes of High and Low ph Buffers 0 0 5 0 5 0 Acetophenone Benzene Toluene Acenaphthene Sequence Column Used: Column: Gei-X 5 µm C8 Dimensions: 50 x. mm Part o.: 00F-5-E0 Step x High ph Flush Procedures Mobile Phase: A: 0 mm Ammonium Bicarbonate ph 0.5 B: Acetonitrile Gradient: 5 % to 95 % B in Hold at 95 % B for e-equilibrate: 5 % B for Flow ate:.5 ml/ Step High ph Testing Isocratic: 0 mm Ammonium Bicarbonate ph 0.5 / Acetonitrile (50:50) Flow ate:.5 ml/ Detection: UV @ 0 nm Step Samples:.. Tetracaine Diphenhydrae x eutral Flush Procedure Mobile Phase: A: Water B: Acetonitrile Gradient: 5 % B for 5 % to 00 % B in Hold at 00 % B for 5 Flow ate:.5 ml/ Step eutral ph Testing Isocratic: Water / Acetonitrile (5:5) Flow ate:.0 ml/ Detection: UV @ 5 nm Step 5 Samples:.... Acetophenone Benzene Toluene Acenaphthene x Low ph Flush Procedure Mobile Phase: A: 0.5 % Formic Acid in Water B: 0.5 % Formic Acid in Acetonitrile, ph.0 Gradient: 5 % to 95 % B in Hold at 95 % B for e-equilibrate: 5 % B for Flow ate:.5 ml/ Step eutral ph Flush epeats epeats for 0 Cycles Phenomenex
} } Gei U.S. Patent o. 7, 5, 7 Polar Bases at High ph, ph 0.5 Gei 9 Waters XBridge 9 5 7 8 0 App ID,5 7 8 0 App ID 0 0 0 0 0 0 Polar Bases (Beta Blockers) at High ph Y-axis normalized for all chromatograms. Dimensions: 50 x. mm Mobile Phase: A: 0 mm Ammonium Bicarbonate ph 0.5 B: Acetonitrile Gradient: A/B (85:5) to (70:0) in 5 to (50:50) in 5, Hold for 5 Flow ate:.5 ml/ Detection: UV @ 0 nm Sample:. Bisoprolol Contaant. Sotalol. Atenolol. Labetalol (Diastereoisomeric Pair) 5. adolol (Diastereoisomeric Pair). Pindolol 7. Metoprolol 8. Bisoprolol 9. Propranolol 0. Alprenolol APPVED HPLC GEMII Polar Bases at Low ph, ph.7 Gei Waters XBridge 7 5 App ID 0, 5 7 App ID 0 8 0 0 8 0 Polar Bases (Antihistaes) in Formic Acid Dimensions: 50 x. mm Mobile Phase: A: 0. % Formic Acid in Water B: 0. % Formic Acid in Acetonitrile Gradient: A/B (90:0) to (50:50) in 0 Flow ate:.5 ml/ Detection: UV @ 0 nm Y-axis normalized for all chromatograms. Sample:. Pyrilae. Tripelennae. Chlorphenirae. Bromphenirae 5. Chloropyrae. Diphenhydrae 7. Loratadine Comparative chromatograms may not be representative of all applications. Phenomenex 7
U.S. Patent o. 7, 5, 7 Gei C8 Increased loading and retention of basic compounds Wide ph stability from - Silica efficiency and mechanical strength Excellent media for process-scale purification Material Characteristics Packing Material Particle Shape/Size (µm) Pore Size (Å) Surface Area (m /g) Carbon Load % End Capping Gei C8 Spherical,5, 0 0 75 TMS Gei C8 USP: L LC/MS Certified Hydrophobicity H-Bonding Aromatic Selectivity Silanol Activity (ph.5) Low see p. 7 for testing probes. ph Stability.0-.0 Particle Size, 5, and 0 μm Phase C8 Application Small Molecules, Basic Compounds Strength Wide ph Stability, High Efficiency High APPVED Batch-to-Batch eproducibility ph Quick Guide Effects of Mobile Phase ph on Acids GEMII HPLC App ID 588 Batch Batch Batch Low ph High ph Acids Above pk a At pk a Below pk a Batch 0 8 Conditions for all separations: Column: Gei 5 μm C8 Dimension: 50 x. mm Part o.: 00F-5-E0 Mobile Phase: 0 mm Ammonium Bicarbonate, ph 0.5/Acetonitrile (50:50) Flow ate:.0 ml/ Detection: UV @ 0 nm Sample:. Pindolol. Metoprolol. Propranolol Effects of Mobile Phase ph on Bases High ph Low ph Below pk a At pk a Bases Above pk a 8 Phenomenex
U.S. Patent o. 7, 5, 7 Virtually o LC/MS Bleed The increased stability of Gei columns provide the ultra-low bleed profile required for high sensitivity LC/MS applications. The advanced TWI Technology used for Gei columns limit phase cleavage and stabilizes the silica surface. When compared against a background MS profile, Gei columns demonstrate virtually no column bleed across a wide mass range. Total Ion Current Extended Column Lifetime The TWI Technology engineering of Gei provide stability and increased column lifetime. Whether used under isocratic or gradient conditions, Gei columns out-perform and outlasts ph stable columns. This is illustrated below. Lifetime and Efficiency Comparison** 9.0e7 8.0e7 7.0e7 Essentially no bleed! 000 000 5.5 times increased column lifetime and still going!.0e7 000 5.0e7 0000.0e7.0e7.0e7 Efficiency () 8000 000 000 Columns fail after 50% loss in efficiency. Gei Fail Line XTerra Fail Line APPVED.0e7 000 0.0 5 7 8 9 0 Column: Gei 5 μm C8 Dimension: 50 x.0 mm Part o.: 00F-5-Y0 Mobile Phase: A: 0.% HCH in Acetonitrile B: 0.% HCH in Water Gradient: 5:95 (A/B) linear to 95:5 over 8, Hold for 5 Flow ate: 0.5 ml/ Detection: API000 Ion Source:ESI/PIM Scan ate:000 m/z/s Scan ange:0-000 Gei C8 Background MS Profile 0 7 0 9 5 8 7 0 9 Days Efficiency and lifetime comparison based on average of two columns each run in parallel. Columns: Gei 5 μm C8 Waters XTerra 5 μm MS C8 Dimensions: 50 x. mm Mobile Phase: Acetonitrile/50 mm Methylpyrrolidine Buffer, ph.5 (50:50) Flow ate: ml/ Detection: UV @ 5 nm Sample: Diphenhydrae Gei C8 Waters XTerra MS C8 HPLC GEMII Chromatographic Comparisons** Gei Z Virtually no silanol interactions! 0 8 0 Tricyclic Antidepressants at eutral ph Dimensions: 50 x. mm Mobile Phase: 0 mm Phosphate buffer ph 7.0/Acetonitrile/ Methanol (0:5:5) Flow ate:.5 ml/ Detection: UV @ 5 nm Sample:. ortriptyline. Imiprae. Amitriptyline. Clomiprae App ID 5595 0 8 0 0 8 0 App ID 50 App ID 559 **The comparative data presented here may not be representative for all applications. Gei is a registered trademark of Phenomenex, Inc. TWI and Two-In-ne Technology are trademarks of Phenomenex, Inc. XTerra is a registered trademark of Waters Corp. ZBAX is a registered trademark of Agilent Technologies. ACE is a registered trademark of Advanced Chromatography Technologies. Phenomenex is not associated in any way with Waters Corp, Agilent Technologies or ACT. Phenomenex 9
U.S. Patent o. 7, 5, 7 Gei C-Phenyl Good selectivity for aromatic compounds Extremely low UV and MS bleed Wide ph stability from - Gei C-Phenyl Hydrophobicity Polarity H-Bonding Aromatic Selectivity Low High Material Characteristics Packing Material Particle Shape/Size (µm) Pore Size (Å) Surface Area (m /g) Carbon Load % End Capping Gei C-Phenyl Spherical,5 0 75 TMS USP: L LC/MS Certified Silanol Activity (ph.5) see p. 7 for testing probes. ph Stability.0-.0 Particle Size and 5 μm Phase Phenyl with C linker Application Aromatic, polar or basic compounds Strength High aromatic selectivity with exceptional peak shape even in neutral conditions. Extremely low bleed phenyl column. eproducible Phenyl Phase Aliphatic Acid Application APPVED GEMII HPLC 0 5 7 8 9 Low Bleed Phenyl Phase Column: Gei 5 µm C-Phenyl Dimensions: 50 x. mm Part o.: 00F--E0 Mobile Phase: 0 mm Phosphate buffer, ph.5/ Methanol (97:) Flow ate:.0 ml/ Detection: UV @ 0 nm Sample:. Tartaric Acid. Acetic Acid. Malic Acid 5. Citric Acid. Lactic Acid. Propionic Acid ne of the weaknesses of traditional phenyl phases has been excessive bleed in UV and MS. Gei C-Phenyl columns demonstrate far lower bleed than many other phenyl columns. Indeed, in this example the Gei C-Phenyl column shows lower bleed than a popular C8 column. Enhanced Performance for Aromatic Compounds Sulfa Drug Application ESLUTI Pursuit 5 µm DiPhenyl Gei 5 µm C-Phenyl s,.0.0 s, 9.8.0 mau 0 80 0 0 0 5 0 5 mau 0 0 0 Phenomenex Gei App ID 590 Varian Pursuit App ID 5908 Intensity cps 9.0e7 8.0e7 7.0e7.0e7 5.0e7.0e7.0e7.0e7.0e7 0.0 Popular C8 Gei C-Phenyl Blank 0 8 0 Dimensions: 50 x.0 mm Mobile Phase: A: 0. % Formic acid in Water B: 0. % Formic acid in Acetonitrile Gradient: 5 % B to 95 % B in 0, then hold 95 % B for Flow ate: 0. ml/ MS Detection: ESI + ion mode, M/Z 00-700 0 0 5 0 5 Sulfathiazole H H S Sulfamerazine H S S H Dimensions: 50 x. mm Mobile Phase: 0. % Formic Acid in Water/ Methanol (70:0) Flow ate:.0 ml/ Detection: UV @ 5 nm Sample:. Sulfathiazole. Sulfamerazine. Sulfamethoxazole The comparative data presented here may not be representative for all applications. Pursuit is a registered trademark of Varian, Inc. CH 70 Phenomenex
U.S. Patent o. 7, 5, 7 Applications Local Anesthetics Column: Gei 5 μm C8 Dimensions: 50 x. mm Part o.: 00F-5-E0 Mobile Phase: 0 mm H HC, ph 0.5 / Acetonitrile (50:50) Flow ate: ml/ Temperature: 5 C Detection: UV @ 0 nm Sample:. Procaine. Lidocaine. Tetracaine Caffeine and Metabolites Column: Gei 5 μm C8 Dimensions: 50 x. mm Part o.: 00F-5-E0 Mobile Phase: A: 0 mm Ammonium acetate, ph 7.5 B: Acetonitrile Gradient: 98: (A/B) linear to 80:0 over 5, Hold for Flow ate: ml/ Temperature: 5 C Detection: UV @ 7 nm Sample:. Xanthine. Theobroe. Theophylline. Caffeine APPVED App ID 999 App ID 50 0 8 0 Clinical Solution Benzodiazepines in Plasma SPE Conditions Sample: 0.5 ml Human Plasma + 0.5 ml Water + 0. ml std (.0 µg/ml in Water) Sorbent: Strata-X 0 mg/ ml tubes Part o.: 8B-S00-TAK Condition: ml Methanol followed by ml Water Wash: ml Water, ml of 0 % Methanol Elution: ml Methanol, evaporated to dryness under gentle flow econstitute: with 00 µl water HPLC Conditions Column: Gei 5 μm C8 Dimensions: 50 x.0 mm Part o.: 00F-5-Y0 Mobile Phase: A: 0. % Formic acid in Water B: 0. % Formic acid in Acetonitrile Gradient: 80:0 (A/B) linear to 0:80 over 8, Hold for Flow ate: 0.8 ml/ Temperature: C Detection: UV @ 5 nm Sample:. Chlordiazepoxide. Flurazepam. xazepam. Clonazepam 5. Temazepam. Diazepam 0 5 0 5 Flavonoids Column: Gei 5 μm C-Phenyl Dimensions: 50 x. mm Part o.: 00F--E0 Mobile Phase: 0. % Formic acid in Water/Methanol (5:55) Flow ate: ml/ Detection: UV @ 5 nm Sample:. Quercetin. Kaempferol. Isorhamnetin App ID 5895 HPLC GEMII 5 Benzodiazepines spiked in human plasma µg/ml benzodiazepine standard Human plasma blank App ID 55, 555, 55 0 8 0 Phenomenex 7
APPVED GEMII HPLC Gei U.S. Patent o. 7, 5, 7 rdering Information μm Microbore, Minibore and arrow Bore Columns (mm) SecurityGuard Cartridges (mm) Phases 50 x.0 0 x.0 0 x.0 50 x.0 00 x.0 50 x.0 50 x.0 00 x.0 50 x.0 x.0* /0pk C8 00B-9-A0 00M-9-B0 00A-9-B0 00B-9-B0 00D-9-B0 00F-9-B0 00B-9-Y0 00D-9-Y0 00F-9-Y0 AJ0-759 C-Phenyl 00B--A0 00A--B0 00B--B0 00D--B0 00F--B0 00B--Y0 00D--Y0 00F--Y0 AJ0-79 /0pk X C8 00M-5-B0 00A-5-B0 00B-5-B0 00D-5-B0 00F-5-B0 00B-5-Y0 00D-5-Y0 00F-5-Y0 AJ0-87 for ID:.0-.0 mm μm Analytical Columns (mm) SecurityGuard Cartridges (mm) Phases 0 x.0 0 x. 50 x. 00 x. 50 x. 50 x. x.0* /0pk C8 00M-9-D0 00A-9-E0 00B-9-E0 00D-9-E0 00F-9-E0 00G-9-E0 AJ0-7597 C-Phenyl 00A--E0 00B--E0 00D--E0 00F--E0 00G--E0 AJ0-795 /0pk X C8 00B-5-E0 00D-5-E0 00F-5-E0 00G-5-E0 AJ0-88 for ID:.-8.0 mm 5 μm Minibore and arrow Bore Columns (mm) SecurityGuard Cartridges (mm) Phases 0 x.0 50 x.0 50 x.0 50 x.0 50 x.0 00 x.0 50 x.0 50 x.0 x.0* /0pk C8 00A-5-B0 00B-5-B0 00F-5-B0 00G-5-B0 00B-5-Y0 00D-5-Y0 00F-5-Y0 00G-5-Y0 AJ0-759 C-Phenyl 00A--B0 00B--B0 00F--B0 00B--Y0 00F--Y0 00G--Y0 AJ0-79 /0pk X C8 00A-5-B0 00B-5-B0 00F-5-B0 00B-5-Y0 00D-5-Y0 00F-5-Y0 00G-5-Y0 AJ0-87 for ID:.0-.0 mm 5 μm Analytical Columns (mm) SecurityGuard Cartridges (mm) Phases 0 x. 50 x. 00 x. 50 x. 50 x. x.0* /0pk C8 00A-5-E0 00B-5-E0 00D-5-E0 00F-5-E0 00G-5-E0 AJ0-7597 C-Phenyl 00A--E0 00B--E0 00D--E0 00F--E0 00G--E0 AJ0-795 /0pk X C8 00B-5-E0 00D-5-E0 00F-5-E0 00G-5-E0 AJ0-88 for ID:.-8.0 mm 5 μm Semi-Prep Columns (mm) SecurityGuard Cartridges (mm) Phases 50 x 0 50 x 0 0 x 0 /pk C8 00F-5-0 00G-5-0 AJ0-7598 C-Phenyl 00G--0 AJ0-7 /pk X C8 00F-5-0 00G-5-0 AJ0-89 for ID: 9- mm For PEP Columns & Bulk Media, see p. 0 For SecurityGuard Holders and Cartridges, see p. For MercuryMS LC/MS Columns, Cartridges, and Cartridge Holders, Inquire. If Gei analytical columns do not provide at least an equivalent separation as compared to a competing column of similar particle size, similar phase and dimensions, send in your comparative data within 5 days and keep the Gei column for FEE. For Gei Capillary HPLC Columns, Guards, and Adapter, contact your Phenomenex technical consultant or local distributor. Axia Packed Preparative Columns (mm) SecurityGuard Cartridges (mm) Phases 50 x. 00 x. 50 x. 50 x. 50 x 0 75 x 0 5 x.** 5 x 0.0 5 μm /ea /ea C8 00B-5-P0-AX 00D-5-P0-AX 00F-5-P0-AX 00G-5-P0-AX 00B-5-U0-AX 00C-5-U0-AX AJ0-78 AJ0-808 C-Phenyl 00D--P0-AX 00F--P0-AX 00G--P0-AX 00C--U0-AX AJ0-78 AJ0-80 5 μm /ea /ea X C8 00B-5-P0-AX 00D-5-P0-AX 00F-5-P0-AX 00G-5-P0-AX 00B-5-U0-AX 00C-5-U0-AX AJ0-870 AJ0-87 0 μm /ea /ea C8 00B--P0-AX 00D--P0-AX 00F--P0-AX 00G--P0-AX 00B--U0-AX AJ0-78 AJ0-808 0 μm /ea /ea X C8 00B-55-P0-AX 00D-55-P0-AX 00F-55-P0-AX 00G-55-P0-AX AJ0-870 AJ0-87 for ID: 8-9 mm 0-9 mm Axia Packed Preparative Columns (mm) continued SecurityGuard Cartridges (mm) Phases 00 x 0 50 x 0 50 x 0 50 x 50 00 x 50 50 x 50 50 x 50 5 x 0.0 5 μm Inquire Inquire Inquire /ea C8 00D-5-U0-AX 00F-5-U0-AX 00G-5-U0-AX 00B-5-V0-AX AJ0-808 C-Phenyl 00D--U0-AX 00G--U0-AX AJ0-80 5 μm Inquire Inquire Inquire /ea X C8 00D-5-U0-AX 00F-5-U0-AX 00G-5-U0-AX 00B-5-V0-AX AJ0-87 0 μm Inquire Inquire Inquire Inquire Inquire Inquire /ea C8 00D--U0-AX 00F--U0-AX 00G--U0-AX 00B--V0-AX 00D--V0-AX 00F--V0-AX 00G--V0-AX AJ0-808 0 μm Inquire Inquire Inquire Inquire Inquire Inquire /ea X C8 00D-55-U0-AX 00F-55-U0-AX 00G-55-U0-AX 00B-55-V0-AX 00D-55-V0-AX 00F-55-V0-AX 00G-55-V0-AX AJ0-87 for ID: 0-9 mm *SecurityGuard Analytical Cartridges require holder, Part o.: KJ0-8 SemiPrep SecurityGuard Cartridges require holder, Part o.: AJ0-70 **PEP SecurityGuard Cartridges require holder, Part o.: AJ0-8 PEP SecurityGuard Cartridges require holder, Part o.: AJ0-877 7 Phenomenex