Core-Shell Technology. 11 HPLC Myths. Uncovered. & Why You Must Have Kinetex Core-Shell HPLC Columns

Transcription

1 Core-Shell Technology 11 HPLC Myths Uncovered & Why You Must Have Kinetex Core-Shell HPLC Columns 2.6

2 HPLC myths have been around since the early 19 s! From the one about the handsome architect who started a chromatography company to the one about the best silica coming from the beaches of Fiji. Here at Phenomenex we believe that you deserve the truth! The whole TRUTH! 214 Phenomenex, Inc. All rights reserved. 2

3 MYTH #1 Fully porous silica is just as good as Core-Shell Technology. Core-Shell Technology No Way! Kinetex Core-Shell Technology produces increased efficiencies over traditional columns on the market, yielding remarkable chromatographic resolution, higher peak capacities, and greater sensitivity, so you can get the most out of every HPLC analysis. Kinetex Core-Shell Fully Porous Less Band Broadening More Band Broadening See how it works at Phenomenex l WEB: 3

4 MYTH #2 One core-shell particle size is all that exists for HPLC. While other column manufacturers add to this myth, we listened to your needs and designed and manufactured two Kinetex core-shell particles to cover methods on your HPLC systems. 2.6 Kinetex Replace 3 and fully porous columns for better performance at low backpressures Enhance LC/MS methods and column lifetime without small particle band broadening concerns Easily achieve high efficiency preparative LC methods Kinetex 2.6 Save time and money with smaller, high performance columns Use HPLC/UHPLC versatility to upgrade methods See more peaks with sub-2 like performance on HPLC and UHPLC systems Other Core-Shell Solutions Kinetex UHPLC Columns Use sub-2 Kinetex particles to stimulate your UHPLC analyses and achieve the highest UHPLC performance possible. Aeris Columns Aeris WIDEPORE and PEPTIDE columns are excellent tools for HPLC analysis of proteins and peptides! solid core solid core solid core

5 MYTH #3 Kinetex columns can t be used on all HPLC systems. Core-Shell Technology That s Crazy! Worldwide compatibility is the truth! Thousands of scientists all around the world are proving this myth false each and every day. Kinetex HPLC columns are absolutely compatible with all HPLC instruments. Kinetex on Agilent 11 Kinetex on Shimadzu Prominence LC-2A Kinetex on Waters Alliance Kinetex on Gilson Preparative HPLC Need help installing your Kinetex column? Waters and Alliance are registered trademarks of Waters Corporation. Agilent is a registered trademark of Agilent Technologies, Inc. Shimadzu and Prominence are registered trademarks of Shimadzu Corporation. Gilson is a registered trademark of Gilson, Inc. Phenomenex is not affiliated with Agilent, Gilson, or Waters. Phenomenex l WEB:

6 MYTH #4 Smaller particles and higher pressures are needed for better HPLC performance. Sometimes stress is not good! High pressure instrumentation can be costly, so why not stick to what you know. By simply dropping in a Kinetex μm for methods like the assay below, you get an impressive enhancement in resolution and sensitivity. Now imagine what this column can do for all your other 3 and μm methods. mau Kinetex C18 Height = 16 mau R S = 2.23 Increased Resolution Increased Sensitivity On Your HPLC System 8 b c 6 a 4 2 b c d App ID min mau Agilent ZORBAX μm XDB-C18 Height = 12 mau a b c R S =.98 b c d App ID 2766 Conditons for both columns Columns: Kinetex μm C18 ZORBAX μm XDB-C18 Dimensions: 2 x 4.6 mm Mobile Phase: A: TFA/Water/THF (:9:1) B: TFA/Acetonitrile/THF (:9:1) Gradient: Time (min) 4 Flow Rate: 1. ml/min Temperature: Ambient Detection: 29 nm Sample: 1. Impurity A 2. Impurity B 3. Impurity C 4. Unidentified. Paroxetine 6. Impurity D % B min Woohoo! No need to change my instrument or conditions! Agilent and ZORBAX are registered trademarks of Agilent Technologies, Inc. (Palo Alto, CA, USA). Phenomenex is not affiliated with Agilent Technologies. Comparative separations may not be representative of all applications. 6

7 MYTH # For great performance you have to sacrifice column lifetime. Core-Shell Technology Alongside LC/UV analyses, the high performance and low pressure of the Kinetex μm make it an excellent tool for LC/MS and LC/MS/MS. Pull out isobaric ions with the high efficiency of Kinetex μm, all at low μm backpressures that promote longer column lifetime and less system strain. 2.6e6 2.4e6 2.2e6 2.e6 1.8e6 1.6e6 1.4e6 1.2e6 1.e6 8.e 6.e 4.e 2.e. 2.6e6 2.4e6 2.2e6 2.e6 1.8e6 1.6e6 1.4e6 1.2e6 1.e6 8.e 6.e 4.e 2.e. Kinetex μm C18 72 Bar min Waters SunFire 3. μm C18 14 Bar min App ID 2771 App ID 2776 Greater resolution at half the pressure Conditons for both columns Columns: Kinetex μm C18 SunFire 3. μm C18 Dimensions: x 2.1 mm Mobile Phase: A: 3 mm Ammonium formate B: Acetonitrile Gradient: Time (min) % B Flow Rate:. ml/min Temperature: Ambient Detection: MS/MS (API 4 ) Sample: 1. DMIP 2. IQx 3. IQ 4. 8-MeIQ. 4,8-DiMe-IQx 6. 7,8-DiMe-IQx 7. Trp-P-2 8. Harman 9. Norharman 1. PhIP 11. AaC 12. MeAaC 13. MeIQ What will your method look like with Kinetex? Find out at: Waters and SunFire are registered trademarks of Waters Corporation. Phenomenex is in no way affiliated with Waters Corporation. Comparative separations may not be representative of all applications. Phenomenex l WEB: 7

8 MYTH #6 Your compound screens are limited. Don t feel trapped! There s a solution! Break away from traditional methods and easily attain fast and comprehensive compound screens with the incredibly high HPLC efficiency of Kinetex 2.6 μm columns. Now you can obtain higher throughput without sacrificing resolution. 4.4e 4.e 3.6e 3.2e 2.8e 2.4e 2.e 1.6e 1.2e 8.e4 4.e4 18 Multi-Class Antibiotics Screening of Meat , , min Antibiotics from 7 different classes App ID 18 Column: Kinetex 2.6 μm C18 Dimensions: x 2.1 mm Part No.: B-4462-AN Mobile Phase: A:.1 % Formic acid in Water B:.1 % Formic acid in Methanol Gradient: Time (min) % B Time (min) Flow Rate:. ml/min Temperature: 4 C Detection: Mass Spectrometer (MS) (3 C) Detector: AB SCIEX API 4 System Note: Analytes spiked at 1 ng/ml Sample: See full list of analytes at % B e6 2.e6 1.8e6 1.6e6 1.4e6 1.2e6 1.e6 8.e 6.e 4.e 2.e Pain Panel Screen Opiates, Benzodiazepines, and Drugs of Abuse in under minutes min App ID 2214 Conditions for all separations: Column: Kinetex 2.6 Biphenyl Dimensions: x 3. mm Part No.: B-4622-Y Mobile Phase: A: Water with.1 % Formic Acid B: Methanol with.1 % Formic Acid Gradient: Time (min) %B Flow Rate:.6 ml/min Temperature: 4 C Detection: MS/MS (AB SCIEX API ) Sample: 1. Morphine 2. Oxymorphone 3. Hydromorphone 4. Amphetamine 22. Fentanyl 23. Buprenorphine 24. Flurazepam 2. PCP. Methamphetamine 26. Carisoprodol 6. MDA 7. Naloxone 8. Codeine 9. 6-MAM 1. Oxycodone 11. MDMA 12. Naltrexone 13. Hydrocodone 14. MDEA 1. Norfentanyl 16. Tramadol 17. Benzoylecgonine 18. Meprobamate 19. Normeperidine 2. Meperidine 27. Propoxyphene 28. Midazolam 29. Sufentanil 3. Norpropoxyphene 31. EDDP 32. Lorazepam 33. Methadone 34. Clonazepam 3. Oxazepam 36. Hydroxyalprazolam 37. Nordiazepam 38. Flunitrazepam 39. Temazepam 4. Alprazolam 41. Diazepam 21. Norbuprenorphine

9 MYTH #7 High solvent use is part of the job. Core-Shell Technology Savings, not waste, is right around the corner! When chromatographic performance improves you can decrease your analysis time and also decrease your overall solvent consumption without compromising your separations. Use short Kinetex 2.6 μm columns to dramatically decrease the solvent consumption in your laboratory and increase sample throughput. Example Method Consumption ml solvent per run! Column: Traditional C18 Dimensions: 2 x 4.6 mm Flow Rate: 1. ml/min App ID min Less Solvent Consumption with Kinetex Column 4 6 < 4 ml solvent per run! Column: Kinetex 2.6 C18 Dimensions: x 2.1 mm Flow Rate:.6 ml/min App ID min Conditions for both columns: Mobile Phase: A: 2 mm Potassium phosphate ph 7 B: Methanol / Acetonitrile (:) A/B (48:2) Temperature: 4 ºC Detection: 24 nm Sample: 1. Tianeptine 2. Desmethyldoxepin 3. Protriptyline 4. Desipramine. Imipramine 6. Amoxapine 7. Doxepin 8. Nortriptyline 9. Amitriptyline 1. Clomipramine How much can your lab be saving?! Comparative separations may not be representative of all applications. Phenomenex l WEB: 9

10 MYTH #8 Scaling HPLC/UHPLC to PREP is not possible with core-shell columns. Transferring high performance HPLC/UHPLC methods to preparative scales becomes seamless with the full scalability of the Kinetex core-shell product line. Start with the analytical particle column (1.3 μm, 1.7 μm, 2.6 μm, or μm) that best suits your instrumentation and needs, and then direct scale to a Kinetex μm Axia packed preparative column. Analytical method Kinetex 2.6 μm XB-C18 mau App ID 2142 Column: Kinetex 2.6 XB-C18 Dimensions: 1 x 4.6 mm Part No.: F-4496-E Mobile Phase: A:.1 % TFA in Water B:.1 % TFA in Acetonitrile Gradient: Linear 8:1 (A/B) to :9 (A/B) over 1 mins Injection Volume: 1 µl Flow Rate: 1 ml/ min Temperature: Ambient Detection: 21 nm Sample: Crude peptide mix min Preparative scale-up and fraction collection Kinetex μm XB-C18 mv FR 1 FR 2 FR 3 FR min App ID 214 Column: Kinetex XB-C18 Axia Packed Dimensions: 1 x 21.2 mm Part No.: F-46-P-AX Mobile Phase: A:.1% TFA in Water B:.1% TFA in Acetonitrile Gradient: Linear 8:1 (A/B) to :9 (A/B) over 1 mins Injection Volume: 1 ml Flow Rate: 2 ml/ min Temperature: Ambient Detection: 21 nm Sample: Crude peptide mix Analytical fraction analysis Kinetex 2.6 μm XB-C18 Fraction 4 Fraction 3 Fraction 2 Fraction App ID 2143 Column: Kinetex 2.6 XB-C18 Dimensions: 1 x 4.6 mm Part No.: F-4496-E Mobile Phase: A:.1 % TFA in Water B:.1 % TFA in Acetonitrile Gradient: Linear 8:1 (A/B) to :9 (A/B) over 1 mins Injection Volume: 1 µl Flow Rate: 1 ml/ min Temperature: Ambient Detection: 21 nm Sample: Purified fractions min 1

11 MYTH #9 High efficiency prep columns don t exist. Core-Shell Technology Then neither do fully electric cars! With narrower peak widths than fully porous columns across every sample load, Kinetex μm Axia packed prep columns give you the capability of increased sample load and higher throughput for vastly improved purification performance and economics. Kinetex C18 Axia Packed mv 6 Increased Sensitivity and Excellent Loadability! min App ID 2774 Waters XBridge C18 Prep OBD mv min App ID 277 Conditions for both columns: Columns: Kinetex μm C18 Axia Packed XBridge C18 Prep OBD Dimensions: x 21.2 mm (Kinetex) x 19 mm (XBridge) Mobile Phase: A: Water with. % Formic acid B: Acetonitrile with. % Formic acid Gradient: Time (min) 8 11 % B 2 Waters and XBridge are registered trademarks and OBD is a trademark of Waters Corporation. Phenomenex is in no way affiliated with Waters Corporation. Comparative separations may not be representative of all applications. Flow Rate: 3 ml/min Temperature: Ambient Detection: 24 nm Sample: 2 mg/ml in DMSO 1. Doxepin (From 1 - mg on-column) 2. Amitriptyline (From 1 - mg on-column) Kinetex Axia Preparative columns are fantastic! I currently use two Kinetex C18 1 x 21.2 mm columns in parallel for high throughput purifications (<1 mg scale), and Kinetex core shell media delivers significantly improved peak shape and lower backpressure compared to many of the industry. I can also analyze quickly my purified fractions with the same core shell phase on my analytical UPLC system. Chris DeVore, Neurocrine Biosciences Phenomenex l WEB: 11

12 MYTH #1 Core-shell columns are not reproducible. Hogwash! Quality is what our products are built on! Every single Kinetex column and Kinetex batch of media undergoes an incredible battery of quality assurance tests to ensure that you are getting the best column every time. Such tests include particle size distribution (both solid core and shell thickness), surface coverage, carbon load, pore diameter distribution, and many other parameters to ensure exceptional reproducibility. C18 Batch-to-Batch Overlay 2 3 Batch 68 R s (2,3) 2. Batch 73 R s (2,3) Batch 7 R s (2,3) 2.6 Kinetex 2.6 C18 App ID 187, 1876, 1877 Batch 68 Kinetex 2.6 C18 Batch 73 Kinetex 2.6 C18 Batch min Conditions same for all batches: Columns: Kinetex 2.6 μm C18 Dimensions: x 4.6 mm Part No.: B-4462-E Mobile Phase: Water / Acetonitrile (6:3) Flow Rate: 1. ml/min Temperature: Ambient Detection: 24 nm Sample: 1. Uracil 2. Hydroxycortisone 3. Cortisone 4. Cortisone acetate. 17-Hydroxyprogesterone Does your current column manufacturer give you the same assurances or results? 12

13 MYTH #11 Guard columns are not useful. R&D 1 Award Recipient Protect your Kinetex core-shell columns from damaging contaminants and microparticulates with the SecurityGuard ULTRA guard cartridge system! Simple to use Extend column lifetime Pressure rated to 2 psi (1378 bar) Fits virtually all manufacturers columns 2.1 to 4.6 mm ID UHPLC Column Protection SecurityGuard ULTRA Guard Cartridge System (Cartridge connected to Holder) Cartridge* Frit Holder Material Chamber Compounds of Interest Contaminants and Particulates *Cartridge schematic not drawn to scale See it in action: If SecurityGuard ULTRA cartridge protection system does not perform as well or better than your current guard cartridge system of similar phase and dimensions, return the product with comparative data within 4 days for a FULL REFUND. Phenomenex l WEB: 13

14 Uncover the performance potential in your lab! With six selectivities, the perfect combination is within reach. Choose from a wide range of phases for greater flexibility in HPLC method development. Kinetex columns come in a variety of stationary phases to cover a full spectrum of applications ranging from acids and bases, to isomers and extremely polar compounds. Si C18 XB- C18 C8 Endcapped C18 phase Increased retention for polar basic compounds Protective isobutyl side chains Increased retention of polar acidic compounds Endcapped C8 phase Less hydrophobic than a C18 phase H 2 ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN H 2 ACN H ACN 2 H ACN 2 H ACN 2 H 2 HO H 2 OH H 2 HO H 2 H 2 O O H 2 Si H 2 OH Si Si HO OH Phenyl- Hexyl HILIC Biphenyl Phenyl - Hexyl Greater retention and separation of aromatic hydrocarbons Unbonded silica phase Increased retention of polar compounds Biphenyl Excellent reversed phase and aromatic retention and selectivity Material Characteristics Packing Material Pore Size (Å) Effective Surface Area (m 2 /g) Effective Carbon Load % USP Classification ph Stability Pressure Stability Kinetex C L * 1 bar** Kinetex XB-C L * 1 bar** Kinetex C L * 1 bar** Kinetex Phenyl-Hexyl L * 1 bar** Kinetex HILIC 1 2 L bar** Kinetex Biphenyl L * 1 bar** * Columns are ph stable from 1.-1 under isocratic conditions. Columns are ph stable under gradient conditions. ** 2.1 mm ID Kinetex columns are pressure stable up to 1 bar. 3. mm and 4.6 mm ID Kinetex 2.6 columns are stable up to 6 bar. 14

15 Kinetex Ordering Information 2.6 SecurityGuard SecurityGuard Columns (mm) ULTRA Cartridges ULTRA Cartridges Phases x 2.1 3/pk x x x x 4.6 3/pk XB-C18 B-46-AN AJ-8782 B-46-E D-46-E F-46-E G-46-E AJ-8768 C18 B-461-AN AJ-8782 B-461-E D-461-E F-461-E G-461-E AJ-8768 C8 B-468-AN AJ-8784 B-468-E D-468-E F-468-E G-468-E AJ-877 Phenyl-Hexyl B-463-AN AJ-8788 B-463-E D-463-E F-463-E G-463-E AJ-8774 Biphenyl B-4627-AN AJ-929 B-4627-E D-4627-E F-4627-E G-4627-E AJ-927 for 2.1 mm ID for 4.6 mm ID SecurityGuard Axia Packed Preparative Columns (mm) PREP Cartridges** Phases x x x x x 21.2 /ea XB-C18 B-46-P-AX D-46-P-AX F-46-P-AX G-46-P-AX AJ-914 C18 B-461-P-AX D-461-P-AX F-461-P-AX G-461-P-AX AJ-914 C8 B-468-P-AX D-468-P-AX F-468-P-AX G-468-P-AX AJ-92 Phenyl-Hexyl B-463-P-AX D-463-P-AX F-463-P-AX G-463-P-AX AJ-9147 for mm ID SecurityGuard Axia Packed Preparative Columns (mm) (continued) PREP Cartridges *** Phases x 3 1 x 3 1 x 3 2 x 3 1 x 3 /ea XB-C18 B-46-U-AX D-46-U-AX F-46-U-AX G-46-U-AX AJ-924 C18 B-461-U-AX D-461-U-AX F-461-U-AX G-461-U-AX AJ-924 C8 B-468-U-AX D-468-U-AX F-468-U-AX G-468-U-AX AJ-9217 Phenyl-Hexyl B-463-U-AX D-463-U-AX F-463-U-AX G-463-U-AX AJ-9216 for 3 mm ID SecurityGuard 2.6 μm Analytical Columns (mm) ULTRA Cartridges Phases 3 x 4.6 x x x x 4.6 3/pk If you are not completely satisfied with Kinetex core-shell columns, send in your comparative data to a similar product with the Kinetex column within 4 days for a FULL REFUND. XB-C18 B-4496-E C-4496-E D-4496-E F-4496-E AJ-8768 C18 A-4462-E B-4462-E C-4462-E D-4462-E F-4462-E AJ-8768 C8 B-4497-E C-4497-E D-4497-E F-4497-E AJ-877 HILIC B-4461-E C-4461-E D-4461-E F-4461-E AJ-8772 Phenyl-Hexyl B-449-E C-449-E D-449-E F-449-E AJ-8774 Biphenyl B-4622-E D-4622-E F-4622-E AJ-927 for 4.6 mm ID SecurityGuard 2.6 μm MidBore Columns (mm) ULTRA Cartridges Phases 3 x 3. x 3. 7 x 3. 1 x 3. 1 x 3. 3/pk XB-C18 A-4496-Y B-4496-Y C-4496-Y D-4496-Y F-4496-Y AJ-877 C18 A-4462-Y B-4462-Y C-4462-Y D-4462-Y F-4462-Y AJ-877 C8 A-4497-Y B-4497-Y C-4497-Y D-4497-Y F-4497-Y AJ-8777 HILIC A-4461-Y F-4461-Y AJ-8779 Phenyl-Hexyl D-449-Y F-449-Y AJ-8781 Biphenyl B-4622-Y D-4622-Y AJ-928 for 3. mm ID SecurityGuard 2.6 μm Minibore Columns (mm) ULTRA Cartridges Phases 3 x 2.1 x x x x 2.1 3/pk XB-C18 A-4496-AN B-4496-AN C-4496-AN D-4496-AN F-4496-AN AJ-8782 C18 A-4462-AN B-4462-AN C-4462-AN D-4462-AN F-4462-AN AJ-8782 C8 A-4497-AN B-4497-AN C-4497-AN D-4497-AN F-4497-AN AJ-8784 HILIC A-4461-AN B-4461-AN C-4461-AN D-4461-AN F-4461-AN AJ-8786 Phenyl-Hexyl A-449-AN B-449-AN C-449-AN D-449-AN F-449-AN AJ-8788 Biphenyl B-4622-AN D-4622-AN AJ-929 for 2.1 mm ID SecurityGuard ULTRA cartridges require holder, Part No.: AJ-9 **PREP SecurityGuard Cartridges require holder, Part No.: AJ-8223 ***PREP SecurityGuard Cartridges require holder, Part No.: AJ-8277 Find Kinetex 1.7 and 1.3 UHPLC analytical dimensions online at: Phenomenex l WEB: 1

16 Core-Shell Technology HPLC Myths Uncovered & Why You Must Have Kinetex Core-Shell HPLC Columns Australia t: f: Austria t: f: Belgium t: (French) t: (Dutch) f: +31 () beinfo@phenomenex.com Canada t: (8) f: (31) info@phenomenex.com Denmark t: f: nordicinfo@phenomenex.com Finland t: f: nordicinfo@phenomenex.com France t: f: franceinfo@phenomenex.com Germany t: f: anfrage@phenomenex.com India t: f: indiainfo@phenomenex.com Ireland t: f: eireinfo@phenomenex.com Italy t: f: italiainfo@phenomenex.com Luxembourg t: +31 () f: +31 () nlinfo@phenomenex.com Mexico t: f: tecnicomx@phenomenex.com The Netherlands t: f: nlinfo@phenomenex.com New Zealand t: f: nzinfo@phenomenex.com Norway t: 81 2 f: nordicinfo@phenomenex.com Puerto Rico t: (8) 41-HPLC f: (31) info@phenomenex.com Sweden t: f: nordicinfo@phenomenex.com United Kingdom t: f: ukinfo@phenomenex.com United States t: (31) 212- f: (31) info@phenomenex.com All other countries: Corporate Office USA t: (31) 212- f: (31) info@phenomenex.com Phenomenex products are available worldwide. For the distributor in your country, contact Phenomenex USA, International Department at international@phenomenex.com BR _W Terms and Conditions Subject to Phenomenex Standard Terms & Conditions, which may be viewed at com/termsandconditions. Trademarks Kinetex is a registered trademark, Aeris, Axia, MidBore, and SecurityGuard are trademarks of Phenomenex. Waters, Alliance, SunFire, XBridge are registered trademarks, OBD is a trademark of Waters Corporation. Agilent and ZORBAX are registered trademarks of Agilent Technologies, Inc. API 4 and API are registered trademarks of AB SCIEX Pte. Ltd. AB Sciex is being used under license. Shimadzu and Prominence are registered trademarks of Shimadzu Corporation. Gilson is a registered trademark of Gilson, Inc. Disclaimer Phenomenex is not affiliated with Agilent Technologies, Gilson, Shimadzu Corporation, or Waters Corporation. Comparative separations may not be representative of all applications. Axia is patented by Phenomenex. U.S. Patent No. 7,674,383 SecurityGuard is patented by Phenomenex. U.S. Patent No. 6,162,362 CAUTION: this patent only applies to the analytical-sized guard cartridge holder, and does not apply to SemiPrep, PREP or ULTRA holders, or to any cartridges. The opinions stated herein are solely those of the speaker and not necessarily those of any company or organization. 214 Phenomenex, Inc. All rights reserved.