LC Column Cleaning and Regeneration

Over time, our liquid chromatography columns will accumulate some contamination, especially if we regularly analyze dirty samples. There are some steps that we can take to keep our columns clean. Further, we can do some limited column regeneration as well, which can often help restore some performance to our columns and extend their lifetime.
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Additional Resources
LC Column Cleaning Recommendations | www.restek.com...
LC Guard Column Selector | www.restek.com...
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Restek Recommended Tools & Parts
LC Columns | www.restek.com...
Guard Cartridges and Hardware | www.restek.com...
Mobile Phase Accessories | www.restek.com...
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[@hanumantharaorao5874]

Nice information

[@pharmasolutionpakistan7668]

Very good effort

[@user-in7kb3ry4n]

good afternoon, thanks for the information provided. Please tell me what we need to do if the input part of the column starts to clog (raptor biphenyl 2.7um)? And why can't I reverse flush this column? (Due to the fact that it fails?)

[@RestekCorp]

You can reverse the 2.7um columns. However, 2.0um and small should NOT be reversed. You can find detailed instructions here www.restek.com/en/technical-literature-library/articles/restek-liquid-chromatography-LC-columns--cleaning-recommendations/

[@ajitjadhav607]

Thnks for information. Kindly explain of each solvents use or role in column regeneration. Example - Why we use IPA, n-Hexane, Methanol etc

[@RestekCorp]

The range of solvents is used to transition from the more aqueous mobile phase that columns are most often used with to a more organic mobile phase that does a better job of removing contaminants from the column. Then you reverse the process to safely return the column to its more aqueous mobile phase.

[@ajitjadhav607]

@@RestekCorp n-Hexane is a non polar solvent then can why we use n hexane in reverse phase column regeneration. is n-hexane affects on C18 or C8 column efficiency. plz explain.

[@pharmasolutionpakistan7668]

Good column

[@bikesh98]

how can i wash my column after analysi of folic acid in folic and iron tablets? Pressure generally rises after analysis...

[@RestekCorp]

Check out this intro to column rinsing kzbin.info/www/bejne/fGXGlYmOos-FrZo

[@hoa4300]

Can you write a detailed program to clean column C18 with this method? Tks Note: I use pharmaceutical analysis with HPLC Dionex Ultimate 3000 autosampler and Water E2695 autosampler. Example: I used Acetolnitril: KH2PO4 0,01M (40:60) pharmaceutical analysis, so How I can clean column C18 with this method?

[@RestekCorp]

We've a set of detailed instructions written up here www.restek.com/technical-literature-library/articles/restek-liquid-chromatography-LC-columns--cleaning-recommendations/

[@MsKievStar]

How about cleaning a BPG Column Diaphragm?

[@SasiKumar-lg1wc]

How to remove the column blockage?

[@anupsharma7831]

Thnks

[@pihupapa125]

Can we use n Hexane for reversed phase column

[@SasiKumar-lg1wc]

Which solvent suitable for c18 columns?

[@chakhonglam2444]

I have an Agilent ZORBAX NH2 and Carbohydrate column which is used for HILIC separation for sugar. Since those 2 columns have not been used for a period of time. Which solvents are suitable to use for cleaning and regeneration?

[@RestekCorp]

For any specific column, we would always recommend referencing the original manufacturer's instructions. Here is some information that may be helpful www.agilent.com/cs/library/datasheets/public/820629-008D.pdf For general information on LC column regeneration, we have a nice blog post www.restek.com/chromablography/resurrecting-an-old-reverse-phase-lc-column--should-i-could-i-how-would-i

[@ovaneschakoyan6584]

what means return to mobile phase? return to organic phase?if this is the case and phase has some amount salts, it is appropriate for final cleaning?

[@RestekCorp]

Yes, you can return to your original mobile phase even if it has some additives.

[@peaksanalyticalhplcgc5734]

Thank you so much for this information , in this video about the washing of reversed phase column, here you have recommended wash with n-Hexane , but hexane is highly non-polar, so is that really safe to wash the C18 column with hexane?, RP HPLC column are quite non-polar specially e.g. C18 and that we are using more often., here i am little worried., looking forward for your kind reply Regards

[@RestekCorp]

Yes, it is safe to use n-hexane to wash a C18 column, though you may need to use a transition solvent like ethanol or isopropanol if you have a highly polar solvent in your column. You can find some detailed instructions here: www.restek.com/Technical-Resources/Technical-Library/General-Interest/general_MIS_0322

[@SasiKumar-lg1wc]

What is the procedure of new column washing method

[@RestekCorp]

For information on conditioning a new column, we cover that here kzbin.info/www/bejne/gIrEfa2Iit18bdU

[@septianadetama7736]

What is the meaning "20 column volume" That presenter mention?

[@RestekCorp]

The instruction to flush with 20 column volumes means to flush the column with an amount of solvent equal to 20 times the volume of the column itself. For example, a 50 mm long column with 3.0 mm ID would have an approximate volume of 0.35 mL. Twenty times that would be 7 mL, so you should flush with about 7 mL of solvent.

[@jalaluddin4478]

Is this general for all vendor column i.e agilent and Supelco.

[@RestekCorp]

This information is pretty universal. That said, there are always exceptions and we'd encourage you to confirm the recommendations for your specific column.

[@HiteshPatel-hd5jw]

Which flow rate to flush the column??

[@RestekCorp]

The correct flow rate varies by column ID. Suggested flow rates would be 25 uL/min for 1.0 mm ID columns, 100 uL/min for 2.1 mm ID columns, 250 uL/min for 3.0 or 3.2 mm ID columns, and 500 uL/min for 4.6 mm ID columns.

[@14394]

How mach time

[@vivabauergaming1132]

@0:49 the presenter mentions that 20 column volumes of each solvent should be flowed through the column for proper flushing and reconditioning. Following the formula shown @ 1:16 will help you estimate your column volume. Divide your column volume by the flow rate you choose to determine the amount of time required for each flushing step.