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If the column was properly flushed after use and stored in 100% organic solvent such as acetonitrile, the column will survive for many years.

Clean the column with a water/organic solvent mixture, using the same content as in the mobile phase containing an ion-pairing reagent. For example, clean the column with 10% acetonitrile in water for at least 30 minutes. Then, clean it with water/acetonitrile = 50/50 for at least 30 minutes.

Symptoms of deterioration are poor peak shape, split peaks, shoulders, loss of resolution, decreased retention times, and high back pressure. These symptoms indicate contaminants have accumulated on the frit or column inlet, or there are voids, channels, or a depression in the packing bed.

We recommend consulting a chemical disposal company with the MSDS for the packing material and any other chemicals that might also be in the column. It may be possible to then flush the column, properly dispose of its contents, and recycle the hardware.

How to Clean an HPLC Column - Reverse Flush Method - YouTube YouTube Start of suggested clip End of suggested clip Easiest way to remove. This is to reverse the flow on your column you can do this by taking out theMoreEasiest way to remove. This is to reverse the flow on your column you can do this by taking out the column. And seeing how the flow direction is currently.

Below are some of the signs to look for that your relationship and your HPLC column lifetime is coming to an end. High Backpressure not reduced by reverse flushing the column. Split Peaks. Loss of resolution. Broad peak shape. Retention time shifts.

Regeneration Procedure for Normal-Phase Silica Column Disconnect the column and reconnect it to the system, with the flow through the column in the reversed direction.Set the flow at 1 mL/min.Flush column with 50 mL of ethyl acetate. Reconnect the column in the proper flow direction.

To backflush a column, take the column, remove it from the system, and connect the outlet to the inlet tubing from the pump. Pump water, acetonitrile or your mobile phase through the column to dislodge the particles that are blocking the frit. Do not connect your column to the detector when you are backflushing.

How to Clean an HPLC Column - YouTube YouTube Start of suggested clip End of suggested clip The method for reverse flush would be to start with 95. Percent water. And 5 methanol for about 10MoreThe method for reverse flush would be to start with 95. Percent water. And 5 methanol for about 10 column. Volumes. Then you do a gradient of 5 to 95 methanol for five column volumes.

5.6 Discard of HPLC / GC column 1 Discard the column if it is not meeting the criteria of Tailing factor, theoretical plates, Resolution, Fronting, Splitting/ damage etc. 5.6. 3 Discarded columns shall be kept separately and affixed to the label as ?Discarded column-Do not Use?.