Setting the Standard for ph Method Development
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1 U.S. Patent o. 7, 56, 67 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 Gemini 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 Gemini column for FEE. Phase X-C8 C6-Phenyl C8 Description The most rugged Gemini 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 TWI (Two-In-ne) Technology During the final stage of silica manufacturing a unique silica-organic layer is grafted to create a completely new composite particle. nce 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 crosslinking. 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. APPVED HPLC Gemini H H H H H H H H H H H H H H S C C C Phenomenex 87
2 U.S. Patent o. 7, 56, 67 Gemini 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 USP: L ph Stability:.. Particle ze: µm, 5 µm, and μm Phase: C8 Application: Small molecules, basic compounds Strength: Most durable ph stable particle LC/MS Certified APPVED Gemini HPLC Material Characteristics Packing Material Particle Shape/ze (µm) Pore ze (Å) Surface Area (m /g) Carbon Load % End Capping Gemini X-C8 Spherical, 5, 75 TMS Gemini X-C8 Tested for Extreme Durability in Changing Mobile Phase ph Column Efficiencies Maintained in High ph Testing for Cycles % of Initial Efficiencies etention in Minutes Diphenhydramine Cycles of Exposure Tetracaine etention Times of Four Probes Maintained in eutral ph Testing for Cycles 6 5 Stable Efficiencies After Exposure to, Column Volumes of High and Low ph Buffers etention Times Maintained After Exposure to, Column Volumes of High and Low ph Buffers Column Testing Cycle Step x High ph Flush Procedures Mobile Phase: A: mm Ammonium Bicarbonate ph.5 B: Acetonitrile Gradient: 5 % to 95 % B in 6 min Hold at 95 % B for min e-equilibrate: 5 % B for min Flow ate:.5 ml/min Step High ph Testing Isocratic: mm Ammonium Bicarbonate ph.5 / Acetonitrile (5:5) Flow ate:.5 ml/min Detection: nm Step Samples:.. Tetracaine Diphenhydramine x eutral Flush Procedure Mobile Phase: A: Water B: Acetonitrile Gradient: 5 % B for min 5 % to % B in min Hold at % B for 5 min Flow ate:.5 ml/min Step eutral ph Testing Isocratic: Water / Acetonitrile (5:65) Flow ate:. ml/min Detection: 5 nm Samples:. Acetophenone. Benzene. Toluene. Acenaphthene Step 5 x Low ph Flush Procedure Mobile Phase: A:.5 % Formic Acid in Water B:.5 % Formic Acid in Acetonitrile, ph. Gradient: 5 % to 95 % B in 6 min Hold at 95 % B for min e-equilibrate: 5 % B for min Flow ate:.5 ml/min Step 6 eutral ph Flush epeats epeats for Cycles 5 5 Sequence Acetophenone Toluene Column Used: Benzene Acenaphthene Column: Gemini X-C8 5 µm Dimensions: 5 x.6 mm Part o.: F-5-E 88 Phenomenex
3 Polar Bases at High ph, ph.5 APPVED, } } Gemini U.S. Patent o. 7, 56, 67 Gemini X-C8 5 µm 9 Waters XBridge 5 µm C App ID 66, App ID 66 min min Y-axis normalized for all chromatograms. Polar Bases (Beta Blockers) at High ph Dimensions: 5 x.6 mm Mobile Phase: A: mm Ammonium Bicarbonate ph.5 B: Acetonitrile Gradient: A/B (85:5) to (7:) in 5 min to (5:5) in 5 min, Hold for 5 min Flow ate:.5 ml/min Detection: nm Sample:. Bisoprolol Contaminant. Sotalol. Atenolol. Labetalol (Diastereoisomeric Pair) 5. adolol (Diastereoisomeric Pair) 6. Pindolol 7. Metoprolol 8. Bisoprolol 9. Propranolol. Alprenolol Polar Bases at Low ph, ph Gemini X-C8 5 µm 7 Waters XBridge 5 µm C8 HPLC Gemini App ID App ID min 6 8 min Y-axis normalized for all chromatograms. Polar Bases (Antihistamines) in Formic Acid Dimensions: 5 x.6 mm Mobile Phase: A:. % Formic Acid in Water B:. % Formic Acid in Acetonitrile Gradient: A/B (9:) to (5:5) in min Flow ate:.5 ml/min Detection: nm Sample:. Pyrilamine. Tripelennamine. Chlorpheniramine. Brompheniramine 5. Chloropyramine 6. Diphenhydramine 7. Loratadine XBridge and Waters are registered trademarks of Waters Corporation. Phenomenex is in no way affiliated with Waters Corporation. Comparative chromatograms may not be representative of all applications. Phenomenex 89
4 U.S. Patent o. 7, 56, 67 Gemini C8 Increased loading and retention of basic compounds Wide ph stability from - lica efficiency and mechanical strength Excellent media for process-scale purification Gemini C8 USP: L ph Stability:.. Particle ze: µm, 5 µm, and μm Phase: C8 Application: Small molecules, basic compounds Strength: Wide ph stability, high efficiency LC/MS Certified Material Characteristics Packing Material Particle Shape/ze (µm) Pore ze (Å) Surface Area (m /g) Carbon Load % End Capping Gemini C8 Spherical, 5, 75 TMS Batch-to-Batch eproducibility ph Quick Guide Effects of Mobile Phase ph on Acids APPVED Low ph Acids Below pk a App ID 588 At pk a Gemini HPLC Batch Batch 6 8 min Batch Conditions for all separations: Column: Gemini 5 μm C8 Dimension: 5 x.6 mm Part o.: F-5-E Mobile Phase: mm Ammonium Bicarbonate, ph.5/acetonitrile (5:5) Flow ate:. ml/min Detection: nm Sample:. Pindolol. Metoprolol. Propranolol Batch High ph Effects of Mobile Phase ph on Bases High ph Low ph Above pk a Below pk a At pk a Bases Above pk a 9 Phenomenex
5 U.S. Patent o. 7, 56, 67 Virtually o LC/MS Bleed The increased stability of Gemini columns provide the ultra-low bleed profile required for high sensitivity LC/MS applications. The advanced Twin Technology used for Gemini columns limit phase cleavage and stabilizes the silica surface. When compared against a background MS profile, Gemini columns demonstrate virtually no column bleed across a wide mass range. Total Ion Current Extended Column Lifetime The Twin Technology engineering of Gemini provide stability and increased column lifetime. Whether used under isocratic or gradient conditions, Gemini columns out-perform and outlasts ph stable columns. This is illustrated below. Lifetime and Efficiency Comparison** 9.e7 8.e7 7.e7 Essentially no bleed! times increased column lifetime and still going! 6.e7 5.e7.e7.e7.e7 Efficiency () 8 6 Columns fail after 5 % loss in efficiency. Gemini Fail Line XTerra Fail Line.e min Column: Gemini 5 μm C8 Dimension: 5 x. mm Part o.: F-5-Y Mobile Phase: A:. % HCH in Acetonitrile B:. % HCH in Water Gradient: 5:95 (A/B) linear to 95:5 over 8 min, Hold for 5 min Flow ate:.5 ml/min Detection: API Ion Source:ESI/PIM Scan ate: m/z/s Scan ange:6- Gemini C8 Background MS Profile Days Efficiency and lifetime comparison based on average of two columns each run in parallel. Columns: Gemini 5 μm C8 Waters XTerra 5 μm MS C8 Dimensions: 5 x.6 mm Mobile Phase: Acetonitrile/5 mm Methylpyrrolidine Buffer, ph.5 (5:5) Flow ate: ml/min Detection: 5 nm Sample: Diphenhydramine Gemini C8 Waters XTerra MS C8 APPVED Chromatographic Comparisons** Gemini 5 µm C8 Å Virtually no silanol interactions! Agilent Technologies Zorbax 5 µm Extend-C8 8 Å HPLC Gemini 6 8 min Tricyclic Antidepressants at eutral ph Dimensions: 5 x.6 mm Mobile Phase: mm Phosphate buffer ph 7./Acetonitrile/ Methanol (:5:5) Flow ate:.5 ml/min Detection: 5 nm Sample:. ortriptyline. Imipramine. Amitriptyline. Clomipramine App ID min 6 8 min Advanced Chromotography Technologies ACE 5 µm C8 Å App ID 5596 App ID 56 **The comparative data presented here may not be representative for all applications. Gemini is a registered trademark of Phenomenex. Twin and Two-In-ne Technology are trademarks of Phenomenex. Waters and XTerra are registered trademarks of Waters Corp. Zorbax is a registered trademark of Agilent Technologies. ACE is a registered trademark of Advanced Chromatography Technologies Limited. Phenomenex is not associated in any way with Waters Corp, Agilent Technologies or ACT. Phenomenex 9
6 U.S. Patent o. 7, 56, 67 Gemini C6-Phenyl Good selectivity for aromatic compounds Extremely low UV and MS bleed Wide ph stability from - Material Characteristics Packing Material Particle Shape/ze (µm) Pore ze (Å) Surface Area (m /g) Carbon Load % End Capping Gemini C6-Phenyl Spherical, 5 75 TMS Gemini C6-Phenyl USP: L LC/MS Certified ph Stability:.. Particle ze: µm and 5 µm Phase: Phenyl with C6 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 Gemini HPLC Low Bleed Phenyl Phase ne of the weaknesses of traditional phenyl phases has been excessive bleed in UV and MS. Gemini C6-Phenyl columns demonstrate far lower bleed than many other phenyl columns. Indeed, in this example the Gemini C6-Phenyl column shows lower bleed than a popular C8 column. Intensity cps 9.e7 8.e7 7.e7 6.e7 5.e7.e7.e7.e7.e Column: Gemini 5 µm C6-Phenyl Dimensions: 5 x.6 mm Part o.: F--E Mobile Phase: mm Phosphate buffer, ph.5/ Methanol (97:) Flow ate:. ml/min Detection: nm Sample:. Tartaric Acid. Acetic Acid. Malic Acid 5. Citric Acid. Lactic Acid 6. Propionic Acid Popular C8 Gemini C6-Phenyl Blank. 6 8 min Dimensions: 5 x. mm Mobile Phase: A:. % Formic acid in Water B:. % Formic acid in Acetonitrile Gradient: 5 % B to 95 % B in min, then hold 95 % B for min Flow ate:.6 ml/min MS Detection: ESI + ion mode, M/Z -7 min Enhanced Performance for Aromatic Compounds Sulfa Drug Application esolution Pursuit 5 µm DiPhenyl Gemini 5 µm C6-Phenyl s,.. s, mau mau Sulfathiazole H H Sulfamerazine S H Phenomenex Gemini 5 μm C6-Phenyl Agilent Technologies Pursuit 5 μm DiPhenyl S S H CH min min Dimensions: 5 x.6 mm Mobile Phase:. % Formic Acid in Water/ Methanol (7:) Flow ate:. ml/min Detection: 5 nm Sample:. Sulfathiazole. Sulfamerazine. Sulfamethoxazole The comparative data presented here may not be representative for all applications. Agilent and Pursuit are registered trademarks of Agilent Technologies, Inc. App ID 59 App ID Phenomenex
7 U.S. Patent o. 7, 56, 67 Applications Mycotoxin Screening Column: Gemini 5 μm C8 Dimensions: 5 x.6 mm Part o.: F-5-E Mobile Phase: A: Water/Methanol (9:) containing 5 mm Ammonium acetate and % Acetic acid B: Water/Methanol (:97) containing 5 mm Ammonium acetate and % Acetic acid Gradient: Time (min) % B 8..5 Flow ate: ml/min Temperature: 5 C Detection: Tandem Mass Spectrometer (MS/MS) (5 C) Detector: AB SCIEX API System Positive Polarity Intensity, cps Intensity, cps Intensity, cps 5.5e5 5.e5.5e5.e5.5e5.e5.5e5.e5.5e5.e5 5.e Vitamin B.6e5.e5.e5 e5.8e5.6e5.e5.e5 e5.8e5.6e5.e5.e5 e5 8e 6e.e.e egative Polarity 6.e 5.e.e.e.e.e Sample:. Lincomycin. Ergometrine. Aflatoxin G. Aflatoxin G 5. Aflatoxin B 6. Ergocryptine 7. Aflatoxin B Phenomenex 8. Ergocryptinine 9. chratoxin A. igirin. Chloramphenicol. β-zearalenol. apamycin Column: Gemini μm C8 Dimensions: x.6 mm Part o.: D-9-E Mobile Phase: A:. % Formic acid in Water B: Methanol Gradient: Time (min) % B Flow ate:.6 ml/min Temperature: C Detection: Electrospray Mass Spec (ESMS) ( C) Detector: AB SCIEX API System Sample:. icotinamide 99. icotinic acid * Mycotoxins and Metabolites in Minutes! min min min 9 App ID 7 App ID 99 Hormones in Drinking Water: EPA Method 59 Intensity, cps Column: Gemini μm X-C8 Dimensions: x. mm Part o.: D-5-B Mobile Phase: A:. % H H in Water B:. % H H in Methanol Gradient: Time (min) % B Time (min) % B TMP and Thiamine mau Column: Gemini μm X-C8 Dimensions: x. mm Part o.: D-5-Y Mobile Phase: A: 5 mm a HP with % methanol (ph 7.) B: 5 mm a HP with 7 % methanol (ph 7.) Gradient: Time (min) % B Time (min) % B Flow ate:.75 ml/min Detection: Fluorescence (Excitation: 75 nm, Emission: 5 nm) (Ambient) Temperature: C Sample:. TMP. Thiamine App ID 87 min Flow ate:. ml/min Temperature: C Detection: Tandem Mass Sepectrometer (MS/MS) ( C) Detector: AB SCIEX API System Sample:. Estriol 8. Ethynylestradiol-d. Estriol-d (IS) 9. 7α-Ethynylestradiol. Bisphenol A-d6. C-Ethynylestradiol (IS). Equilin. Androstenedione 5. Estrone. Testosterone-d 6. Beta-estradiol. Testosterone 7. C6-estradiol (IS).e5.8e5.6e5.e5.e5.e5.8e5.6e5.e5.e5.e5.8e5.6e5.e5.e5.e5 8.e 6.e.e.e., min , App ID 99 APPVED HPLC Gemini 9
8 APPVED Gemini HPLC Gemini U.S. Patent o. 7, 56, 67 rdering Information μm Microbore, Minibore and arrow Bore Columns (mm) SecurityGuard Cartridges (mm) Phases 5 x. x. x. 5 x. x. 5 x. 5 x. x. 5 x. x.* /pk C8 B-9-A M-9-B A-9-B B-9-B D-9-B F-9-B B-9-Y D-9-Y F-9-Y AJ-7596 C6-Phenyl B--A A--B B--B D--B F--B B--Y D--Y F--Y AJ-79 /pk X-C8 M-5-B A-5-B B-5-B D-5-B F-5-B B-5-Y D-5-Y F-5-Y AJ-867 for ID:.-. mm μm Analytical Columns (mm) SecurityGuard Cartridges (mm) Phases x. x.6 5 x.6 x.6 5 x.6 5 x.6 x.* /pk C8 M-9-D A-9-E B-9-E D-9-E F-9-E G-9-E AJ-7597 C6-Phenyl A--E B--E D--E F--E G--E AJ-795 /pk X-C8 B-5-E D-5-E F-5-E G-5-E AJ-868 for ID:.-8. mm 5 μm Minibore and arrow Bore Columns (mm) SecurityGuard Cartridges (mm) Phases x. 5 x. 5 x. 5 x. 5 x. x. 5 x. 5 x. x.* /pk C8 A-5-B B-5-B F-5-B G-5-B B-5-Y D-5-Y F-5-Y G-5-Y AJ-7596 C6-Phenyl A--B B--B F--B B--Y F--Y G--Y AJ-79 /pk X-C8 A-5-B B-5-B F-5-B B-5-Y D-5-Y F-5-Y G-5-Y AJ μm Analytical Columns (mm) SecurityGuard Cartridges (mm) Phases x.6 5 x.6 x.6 5 x.6 5 x.6 x.* /pk C8 A-5-E B-5-E D-5-E F-5-E G-5-E AJ-7597 C6-Phenyl A--E B--E D--E F--E G--E AJ-795 /pk X-C8 B-5-E D-5-E F-5-E G-5-E AJ-868 for ID:.-8. mm 5 μm Semi-Prep Columns (mm) SecurityGuard Cartridges (mm) Phases 5 x 5 x x /pk C8 F-5- G-5- AJ-7598 C6-Phenyl G-- AJ-956 /pk X-C8 F-5- G-5- AJ-869 for ID: 9-6 mm Axia Packed Preparative Columns (mm) For PEP Columns & Bulk Media, see p. 5 For SecurityGuard Holders and Cartridges, see p. 5 For MercuryMS LC/MS Columns, Cartridges, and Cartridge Holders, Inquire. If Gemini 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 Gemini column for FEE. for ID:.-. mm For Gemini Capillary HPLC Columns, Guards, and Adapter, contact your Phenomenex technical consultant or local distributor. SecurityGuard Cartridges (mm) Phases 5 x. x. 5 x. 5 x. 5 x 75 x 5 x.** 5 x. 5 μm /ea /ea C8 B-5-P-AX D-5-P-AX F-5-P-AX G-5-P-AX B-5-U-AX C-5-U-AX AJ-786 AJ-88 C6-Phenyl D--P-AX F--P-AX G--P-AX C--U-AX AJ-957 AJ μm /ea /ea X-C8 B-5-P-AX D-5-P-AX F-5-P-AX G-5-P-AX B-5-U-AX C-5-U-AX AJ-87 AJ-87 μm /ea /ea C8 B-6-P-AX D-6-P-AX F-6-P-AX G-6-P-AX B-6-U-AX AJ-786 AJ-88 μm /ea /ea X-C8 B-55-P-AX D-55-P-AX F-55-P-AX G-55-P-AX AJ-87 AJ-87 for ID: 8-9 mm -9 mm Axia Packed Preparative Columns (mm) continued SecurityGuard Cartridges (mm) Phases x 5 x 5 x 5 x 5 x 5 5 x 5 5 x 5 5 x. 5 μm Inquire Inquire Inquire /ea C8 D-5-U-AX F-5-U-AX G-5-U-AX B-5-V-AX AJ-88 C6-Phenyl D--U-AX AJ μm Inquire Inquire Inquire /ea X-C8 D-5-U-AX F-5-U-AX G-5-U-AX AJ-87 μm Inquire Inquire Inquire Inquire Inquire Inquire /ea C8 D-6-U-AX F-6-U-AX G-6-U-AX B-6-V-AX D-6-V-AX F-6-V-AX G-6-V-AX AJ-88 μm Inquire Inquire Inquire Inquire Inquire Inquire /ea X-C8 D-55-U-AX F-55-U-AX G-55-U-AX B-55-V-AX D-55-V-AX F-55-V-AX G-55-V-AX AJ-87 for ID: -9 mm *SecurityGuard Analytical Cartridges require holder, Part o.: KJ-8 SemiPrep SecurityGuard Cartridges require holder, Part o.: AJ-7 **PEP SecurityGuard Cartridges require holder, Part o.: AJ-8 PEP SecurityGuard Cartridges require holder, Part o.: AJ Phenomenex
Setting the Standard for ph Method Development
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
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