User Manual
Contents
1. 2 1x30mm 5 um 120A 111365 2103 4 6x250mm 5 wm 120A 111365 4625 4 6x150mm 5 um 120A 111365 4615 4 6x100mm 5 wm 120A 111365 4610 4 6x50mm 5 um 120A 11365 4605 7 8x250mm 5 um 120A 111365 7825 10 0x250mm Sum 120A 111365 10025 21 2x250mm 5 pm 120A 111365 21225 21 2x150mm 5 um 120A 111365 21215 21 2x50mm 5 um 120A 111365 21205 7 8x250mm 10 um 120A 111369 7825 10 0x250mm 10 um 120A 111369 10025 21 2x250mm 10 pm 120A 111369 21225 21 2x150mm 10 pm 120A 111369 21215 21 2x50mm 10 um 120A 111369 212052. silica packing which allows exceptional high stability Such high stability allows GP Phenyl extremely suitable for validation of various analytes The unique mono functional bonding chemistry for GP Pheny avoids the formation of multiple layers Such uniform stationary phase allows the separation to achieve high selectivity and high efficiency A typical test chromatogram for quality control is shown here for a 4 6x250mm GP Pheny column Compared with GP C18 phase GP Phenyl has relatively lower hydrophobicity The high efficiency and less hydrophobicity of GP Phenyl phase make it very suitable for separating compounds with a wide range of hydrophobicity It is highly recommended for separating the compounds which are too strongly retained on C18 phases Safety Precaution GP Phenyl columns are normally operated under high pressure Loose connections will cause leaking of organic solvents and injected samples all of which should be considered as the hazards In the case of leaking proper gloves should be worn for handling the leaked columns When open the columns proper protections should be used to avoid inhalation of the small silica particles Column GP Phenyl Sum 4 6x250mm Mobile phase 60 Methanol 40 H O Detection UV 254 nm Injection volume 2 uL Temperature Ambient Compounds 1 Aniline 0 1 V V 2 Anisole 0 2 V V 3 Toluene 0 5 V V 2 0 60 0 50 0 40 3 2 lt 0 30 1 0 2
3. 0 0 10 0 00 JUL rT 0 0 2 0 4 0 6 0 8 0 10 0 12 0 Min Column Installation and Operation When column is shipped or not in use it is always capped at both ends When install the column to the system first remove the end caps Make the flow direction as marked on the column Unless a user has special purpose to reverse the flow direction for example removal of the inlet pluggage follow the flow direction as labeled Column connections are an integral part of the chromatographic process If ferrules are over tightened not set properly or are not specific for the fitting leakage can occur Set the ferrules for column installation to the HPLC system as follows a Place the male nut and ferrule in order onto a 1 16 o d piece of tubing Be certain that the wider end of the ferrule is against the nut b Press tubing firmly into the column end fitting Slide the nut and ferrule forward engage the threads and fingertighten the nut c While continuing to press the tube firmly into the endfitting use a 1 4 wrench to tighten the nut 90 degrees past fingertightness d Repeat this coupling procedure for the other end of the column New GP Phenyl columns are shipped in a mixture of methanol or acetonitrile and water During stocking and shipping the silica packing could be dried out It is recommended that 10 20 column volumes of pure organic solvents such as methanol acetonitrile be purged to activate the column
4. Flush the column with your mobile phase with gradual increasing the flow rate from 0 1 mL min to your operation condition until the baseline is stable If the column backpressure and baseline fluctuate this might be due to the air bubbles trapped inside the column Flush the column with higher flow rate for 2 5 minutes for example 2 mL min for 4 6x150mm Samples and Mobile Phases To avoid clogging the column all samples and solvents including buffers should be filtered through 0 45 um or 0 2 um filters before use GP Pheny bonded stationary phase is nonpolar in nature It is recommended that the mobile phase be a mixture of organic solvent such as methanol or acetonitrile and water even though GP Pheny can tolerate aqueous buffers as mobile phases Always degas the mobile phase A simple way for degassing is to sonicate it for 5 minutes under water pumped vacuum Gradient elution methods for GP Phenyl columns often begin with 5 methanol or acetonitrile as the initial mobile phase Column Care PH Avoid use of the column below pH 2 or above 9 Higher pH will dissolve silica creating defects of bonding that causes separation efficiency loss and retention time change The optimum performance and operation for longest lifetime are at pH 2 8 5 Pressure Even though GP Phenyl can operate at pressure up to 5 000 psi the normal operation is usually under 3 000 psi Continuous use at high pressure may eventually damage the column as well
5. Sepax Technologies Inc Delaware Technology Park 5 Innovation Way Newark DE 19711 USA Phone 302 366 1101 Fax 302 366 1151 Toll Free 1 877 SEPAX US www sepax tech com Sepax GP Phenyl Column Manual Column Information Utilizing highest purity and enhanced mechanical stability silica and pure bonding reagents GP Phenyl bonded phases have been innovatively and specially designed to ensure maximum mono functional coverage and full end capping which leads to carbon content as high as 11 0 The chemistry of monolayer formation and end capping is completely controlled that results in very reliable column to column reproducibility The maximum surface coverage allows GP Phenyl to have exceptional stability The uniform spherical GP Phenyl particles have a nominal surface area of 300 m g with a controlled pore size of 120A GP Phenyl columns are packed with a proprietary slurry technique to achieve uniform and stable packing bed density for maximum column efficiency GP Pheny packing materials are bonded with phenyl groups that lead to fairly high hydrophobicity GP Phenyl columns have great selectivity and peak symmetry with fairly high retention for separations of acidic neutral and basic organic compounds such as drugs peptides organic acids GP Phenyl columns are especially designed for separation of various organic compounds with ring structure Column Stability and Performance GP Phenyl uses full coverage bonded
6. as the pump Since the pressure is generated by the flow rate The maximum flow rate is limited by the backpressure It is expected that the backpressure might gradually increase with its service A sudden increase in backpressure suggests that the column inlet frit might be plugged In this case it is recommended that the column be flushed with reverse flow in an appropriate solvent Temperature The maximum operating temperature is 60 C Continuous use of the column at higher temperature gt 75 C can damage the column especially under high pH gt 8 5 Storage When not in use for extended time do not allow water or aqueous buffer to remain in the column Remove any aqueous buffers by washing with at least 20 30 column volumes of 50 methanol or acetonitrile aqueous solution followed by 20 30 column volumes of the pure solvent such as acetonitrile Each column is shipped with two removable end plugs To prevent the drying of the column bed seal both ends of the column with the end plugs provided GP Phenyl Products ID x Length Particle Pore size size P N 2 1x150mm 3 um 120A 111363 2115 2 1x100mm 3 um 120A 111363 2110 2 1x50mm 3 um 120A 111363 2105 2 1x30mm 3 um 120A 111363 2103 4 6x250mm 3 um 120A 111363 4625 4 6x150mm 3 um 120A 111363 4615 4 6x100mm 3 um 120A 11363 4610 4 6x50mm 3 um 120A 11363 4605 2 1x250mm 5 um 120A 111365 2125 2 1x150mm 5 um 120A 111365 2115 2 1x100mm 5 um 120A 111365 2110 2 1x50mm 5 um 120A 111365 2105
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