Gemini-NX Performance paired with Axia Technology

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1 Si Si Si Si Si Si Si C Si Si Si Si Si Gemini-NX Performance paired with Axia Technology

2 Gemini -NX Axia Packed Technology Improves Yield, Throughput, and Column Lifetime Ideal Purification Platform While reversed phase separation using HPLC remains one of the most efficient ways of obtaining large-scale purification, several limitations can impact overall productivity: Column loading capacity Media durability Column lifetimes By choosing Gemini-NX in Axia packed preparative columns, these limitations can be overcome; purification yield and throughput can significantly increase. An Equation for Success The success begins with the Gemini-NX media. Through a patentpending process of grafting a network of ethane crosslinking on a pure silica core, a new type of media is created that boasts high column efficiencies, strong hydrophobic selectivity, and a durable ph stability of -. Axia technology complements the package with a self-contained axialcompression piston that locks the media bed in place and eliminates premature column failure due to bed collapse, resulting in a high efficiency, long lasting preparative column. Together, these two technologies complete the equation for increasing productivity for preparative purifications. 009 Phenomenex, Inc. All rights reserved.

3 Increase Sample Loading and Throughput with High ph ph Selectivity Increase loading capacity Sharpen peak shapes Increase productivity Most pharmaceutical compounds are ionizable (either acidic or basic). The ionization state of a compound greatly affects its reversed phase retention. When compounds are ionized, they exhibit less retention. When compounds are neutral, they exhibit greater retention. The ionization state of a compound can be controlled through the mobile phase ph, simply by determining the pk a and adjusting the ph accordingly: For bases, adjust mobile phase ph units above the pk a to neutralize the charge* For acids, adjust the mobile phase ph units below the pk a to neutralize the charge* Because the compound ionization state significantly impacts its reversed phase selectivity, preparative purifications can be optimized to increase yields. When the basic compounds shown in the separation below (diphenhydramine; pk a 8. and propranolol; pk a 9.0) are run under acidic conditions, reversed phase selectivity is minimal, due to the decrease in overall hydrophobicity of the solutes. By increasing the mobile phase ph two units above their respective pk a values to 0., both bases are neutralized increasing reversed phase selectivity, resulting in greater overall resolution and increased sample loading. Low ph Impact of ph on Selectivity* using a Gemini -NX Axia packed column ph.0 High ph ph 0.. Diphenhydramine. Propranolol. Diphenhydramine. Propranolol 0.4 mg in 40 µl App ID mg in 40 µl App ID mg in 40 µl App ID mg in 40 µl App ID min 0 4 min At low ph, when the sample mass is increased to 04 mg, the resolution between the compounds is lost since the two compounds elute so close together. Increasing ph allowed higher mass loading, reducing the number of purification cycles required. The increased resolution at the higher ph also resulted in higher purity and yield for desired basic compounds. * For more detailed information, request technical note TN-00, Impact of ph on the Purity and Yield for Preparative Separations.

4 Gemini -NX ffers Extreme Durability and Robustness in Ever-Changing Environments Rugged Media In today s demanding environments, columns need to deliver more. Consistent performance and long lifetime in the presence of harsh conditions is a requirement for columns that are used routinely for method development or in open-access systems. Gemini-NX columns have been subjected to the most extreme conditions (shown below) to demonstrate their place as your number one choice for reversed phase methods, analytical to preparative. Gemini-NX Tested for Extreme Durability in Changing Mobile Phase ph Column Efficiencies Maintained in High ph Testing for 0 Cycles Column Testing Cycle % of Initial Efficiencies Diphenhydramine Tetracaine Stable Efficiencies After Exposure to 0,000 Column Volumes of High and Low ph Buffers Cycles of Exposure Retention Times of Four Probes Maintained in Neutral ph Testing for 0 Cycles 6 Step. 4x High ph Flush Procedures Mobile Phase: A: 0 mm Ammonium Bicarbonate ph 0. B: Acetonitrile Gradient: % to 9 % B in 6 min Hold at 9 % B for min Re-equilibrate: % B for min Flow Rate:. ml/min Step. High ph Testing Isocratic: 0 mm Ammonium Bicarbonate ph 0. / Acetonitrile (0:0) Flow Rate:. ml/min Detection: 0 nm Samples:. Tetracaine. Diphenhydramine Step. x Neutral Flush Procedure Mobile Phase: A: Water B: Acetonitrile Gradient: % B for min % to 00 % B in min Hold at 00 % B for min Flow Rate:. ml/min Retention in Minutes Acetophenone Benzene Retention Times Maintained After Exposure to 0,000 Column Volumes of High and Low ph Buffers Sequence Toluene Acenaphthene Step 4. Neutral ph Testing Isocratic: Water / Acetonitrile (:6) Flow Rate:.0 ml/min Detection: 4 nm Samples: Acetophenone Benzene Toluene Acenaphthene Step. 4x Low ph Flush Procedure Mobile Phase: A: 0. % Formic Acid in Water B: 0. % Formic Acid in Acetonitrile, ph.0 Gradient: % to 9 % B in 6 min Hold at 9 % B for min Re-equilibrate: % B for min Flow Rate:. ml/min Step 6. Neutral ph Flush Repeats Column Used: Column: Gemini-NX µm C8 Dimensions: 0 x 4.6 mm Part No.: 00F-444-E0 Repeats for 0 Cycles 4

5 ver 000 Cycles with No Loss of Performance Long Column Lifetimes What happens when you combine the durable Gemini -NX media with the advanced technology of Axia packing? You have a preparative column solution that can provide analytical-like efficiencies over extended periods of use. With less frequent column changes, costs per sample decrease and overall productivity increases. Gemini-NX ffers Robust Media for Extreme Conditions at ph 0. (40 ºC) % Initial N vs. Exposure Time (min) in High ph Column: Gemini-NX µm C8 Axia packed Dimension: 00 x. mm Part No.: 00D-444-P0-AX Mobile Phase: A: 0. % Ammonium Hydroxide B: 0. % Ammonium Hydroxide in Acetonitrile Gradient: A/B (9:) to (:9) in 7 min, equilibration in min Flow Rate: 0 ml/min Temperature: 40 C These columns were tested daily in the following isocratic mode: Mobile Phase: Acetonitrile/Water 6/ (v/v) Flow Rate: 0 ml/min Samples: Uracil Acetophenone Benzene Toluene Naphthalene APP ID 7994 CYCLE min CYCLE CYCLE min min Y-axis normalized for all chromatograms.

6 What is Axia Technology? Why Do I Need It? Dependable Axia Technology Benefits: Eliminates media collapse for longer lifetimes Drastically decreases column variability Achieves proper packing density and uniformity Axia technology is beautiful, both on the outside and on the inside. It s an improved, more repeatable packing process that has been recognized by the scientific community as a unique, innovative technology with numerous benefits. The Axia packing process provides preparative chromatographers significant improvements in column lifetime, efficiencies, and peak symmetries that are on par with analytical separations. verall column-to-column consistency is also dramatically improved. With Axia packed columns, the ideal sorbent density is custom calculated and automated for each chemistry and column size. Computer control of the entire process ensures the proper sorbent density and uniformity for an excellent performing preparative column every time. No other company offers this packing technology. 006 R&D 00 Award Recipient We invite you to view the award winning packing process at: Calculating your scaling factor Scaling a separation to meet preparative demands requires the redimensioning of chromatographic parameters such as flow rate/column ID/sample load. When only the column ID is changing: SF = (d /d ) SF= Scaling factor d = diameter of starting column d = diameter scaling to When both ID and length are changing: SF = (d /d ) * (L /L ) L = length of starting column L = length you are scaling to Please us at for further information on large scale preparative and bulk materials. Length (mm) Internal Diameter (mm) 4.6 mm. mm 0 mm 0 mm 0 mm mm mm mm mm Flow Rate (typical to high) (ml/min) Estimating Sample Load (mg) for 4.6 mm ID column to Axia Preparative Columns (. - 0 mm ID) Sample Load (mg) 6

7 SecurityGuard Extends Column Lifetime Protect your Investment Notice in the study below how the column would have been fouled under the aggressive sample loading at 40 injections. With Security- Guard prep the inexpensive cartridge was ruined while the integrity of the prep column was maintained. SecurityGuard prep offers even more injections per column! Forced Degradation Study APP ID Injection : Axia Packed column with SecurityGuard PREP Cartridge 0 4 min Injection 40: Axia Packed column with SecurityGuard PREP Cartridge 40 Time to change the PREP cartridge Conditions Column: Luna 0 µm C8() Axia packed Dimension: 0 x. mm Part No.: 00B-4-P0-AX AJ0-870 (SecurityGuard Cartridge) Mobile Phase: A: 0. % TFA in Water B: 0. % TFA in Water/Acetonitrile (:7) Gradient: Linear 9:7 (A/B) to 00 % B over minutes Injection Volume: 40 µl Flow Rate: 60 ml/min Temperature: Ambient Detection: 70 nm Sample:. Nadolol. Metoprolol. Propranolol min Injection 4: Axia Packed column after removing SecurityGuard PREP cartridge 4 0 riginal column performance maintained with SecurityGuard PREP 4 min If Axia packed columns do not provide LNGER LIFETIME when used with SecurityGuard PREP, as compared to a competing column of the same particle size, phase and dimensions, send in your comparative data and the column within 4 days for a full refund. Supplying Your Laboratory rdering Information μm Axia Packed Preparative Columns (mm) SecurityGuard Cartridges 0 x. 00 x. 0 x. 0 x. 0 x 0 00 x 0 0 x 0 0 x 0 x. ** x 0.0 Phase C8 00B-444-P0-AX 00D-444-P0-AX 00F-444-P0-AX 00G-444-P0-AX 00B-444-U0-AX 00D-444-U0-AX 00F-444-U0-AX 00G-444-U0-AX AJ0-870 AJ0-87 for ID: 8-9 mm 0-49 mm 0 μm Axia Packed Preparative Columns (mm) SecurityGuard Cartridges 0 x. 0 x 0 0 x 0 00 x 0 0 x 0 0 x 0 x. ** x 0.0 Phase C8 00G-44-P0-AX 00G-44-U0-AX 00B-44-V0-AX 00D-44-V0-AX 00F-44-V0-AX 00G-44-V0-AX AJ0-870 AJ0-87 for ID: 8-9 mm 0-49 mm ** 0-49 mm ID PREP SecurityGuard Cartridges Require Holder, Part No.: AJ0-877 PREP SecurityGuard Cartridges require holder, Part No.: AJ

8 Australia t: f: Austria t: f: Belgium t: + (0) f: + (0) Canada t: (800) 4-68 f: (0) Denmark t: f: France t: f: Germany t: f: Ireland t: f: Italy t: 0 67 f: 0 67 Luxembourg t: + (0) f: + (0) Netherlands t: f: New Zealand t: f: Puerto Rico t: (800) 4-HPLC f: (0) United Kingdom t: f: All other countries: Corporate ffice USA t: (0) -0 f: (0) Phenomenex products are available worldwide. For the distributor in your country, contact Phenomenex USA, International Department at BR640908_I Trademarks Luna and Gemini are registered trademarks of Phenomenex, Inc. Axia, ph-lc, and SecurityGuard are trademarks of Phenomenex, Inc. Disclaimer Subject to Phenomenex Standard Terms & Conditions, which may be viewed at Phenomenex, Inc. All rights reserved. 8

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