hplc column care and maintenance
50 Procedure HPLC Column Management General Notes Precautions during handling of HPLC Column. Rinse column remove detector from flow path Eliminate column contamination and plugged packing high molecular weightadsorbed compounds precipitate from sample or buffer Back flush column may clear plugged column inlet frit.
Good Laboratory Practice For Hplc Good Laboratory Practice Cleaning Techniques Cleaning
Flush several volumes of 100 methanol or similar through the HPLC system after water washing a prior to shutting down.
. General Care and Use of Supelco TM HPLC Columns SUPELCO Bellefonte PA T794165E 1998 Sigma-Aldrich Co. Use at least 5 10 volumes of equilibriating mobile phase Before storing rinse the column to remove any residual buffer or ion- pairing agents. Disconnect the column from the HPLC unit and store in an allocated place.
Care Maintenance and Troubleshooting ofTroubleshooting of HPLC Columns Columns and Consumables Edward Kim Apppp glications Engineer January 17 2008 Page 1 Goals for this presentation. The general procedure of column conditioning is as follows. Run linear gradient system from 100 aqueous mobile phase to 100 organic mobile phase up to 2-3 column volume at the identical flow rate as above.
Maintenance of HPLC Columns. Use HPLC grade solvents to prepare the mobile phase. Install the column appropriately when you replace or change it.
Care and Maintenance of HPLC Columns. This HPLC column has been extensively tested and inspected to. Never expose the HPLC column to the pH below or above the pH tolerance limit specified by the manufacturer.
If metal ions are part of the contaminants it may be useful to wash the column with 005 M EDTA to help in solubilizing them. 8515 vv and flush. Initial setup of your LC system is very important to ensure column performance.
Acetonitrile with a maximum of 50 water content. Because of the long storage time it is easy to dry the column. If Column pressure remains high.
Confirm the performance of new column supplied by the vendor. Disconnect the tubing from the column and reattach the inlet tubing to the outlet of the column. Clean pump Injector loop and syringe with hot water in.
Used in the range of low pressure 14 709975kPa the column can maintain high efficiency for a long period of time. 6 The actual operating pressure of the HPLC column should be lower than the pressure during filling and it is recommended to be less than half of the upper limit pressure. Avoid drastic changes in the composition or proportion of the mobile phase.
Generally it does not exceed 20593965 2941995 kPa. Each column is maintained every two months especially for columns that are used less frequently. Always use HPLC grade solvents and water for analysis.
The column is packed with porous particles and its made of polymer and is bounded by a thin layer of silica and polystyrene. This is essential to prevent degradation or buffer salt deposition on drying. Increase the flow rate gradually over a period of 2-5 minutes.
Protect the column from bumps knocks and mechanical shock. Wash the column with a minimum of 30 ml of 25 M acetic acid. Maintenance such as column washing replacement of a guard column or filter or cleaning of an inlet frit.
Wash the column with about 3 column volumes of aqueous mobile Phase with a low flow rate. How to protect LC column Degassed solvents water and mobile phase should be used. Change the mobile phase to a mixture of 001 N H 2 SO 4.
Try to protect the HPLC column with a pre-column or guard column. Explore the reasons for these column problems. Introduce the most commonly observed column related problems in HPLC.
Afterwards the column must be washed thoroughly with water before going back to the initial composition of the mobile phase. Washing of L17 column. The pH range of most HPLC columns is 2-8 try not to exceed the range.
For columns that have really failed make a record of decommissioning do not discard them at will and store them specially. Early in our history our coatings were developed to offer solutions for difficult process sampling and GC sampling applications. Each column is a precision instrument which if cared for properly will provide significant performance lifetime and repeatability.
Lines primed no dry lines or bubbles 3. Make a maintenance record every time. In the early years the Chromatography work carried out by the glass column with the diameter 01 to 05 cm and length is 50 to 500 cm now a day HPLC columns are made of stainless.
17 rows To rejuvenate column performance follow the reverse flow column flush procedure below. Consistent pressures Flush LC system pump and line with mobile phase HPLC grade and miscible with solvents that column is shipped in. Direct the open end of the column over a waste container.
It is designed to help alleviate problems before they occur and covers a range of topics. This column care guide is designed with these factors in mind allowing you to extract the utmost from your HPLC column. Thats why HPLC column care and robust sampling system maintenance is important to achieving reliable HPLC test results and bio-inertness.
A sudden increase in. Storing silica-based HPLC columns for an extended period of time may be done without buffer by applying an organic solvent eg. Always use guard column to protect from impurities and other foreign particulates.
Ensure that your LC system is ready. Aeris AF pak Aqua Asahipak AX pak BioSep-SEC-S Bondclone Ceramospher Chiral CD-Ph Chirex Clarity Columbus Cosmosil Develosil EZfaast Gemini Hypercarb HyperClone Hypersil IB-Sil Inertsil Jupiter Kinetex Kromasil KrudKatcher LiChrosorb LiChrospher Luna Maxsil Nucleodex Nucleosil-Chiral Nucleosil OH pak Onyx OR pak Other Hamilton LC Other. For storage over several days a buffer-containing eluent is recommended to rinse the column with water to prevent microbial growth.
Seals lines injector clean 2.
