I am trying to analyze lipase using the reverse phase-HPLC C18 column. In the literature, 0.1 % TFA was and 60% ACN was used. Can I use TCA in place of TFA?
If any other methods are available to analyze lipase please suggest me.
Thank you.
- "Can I use TCA instead of TFA" ?
Maybe. If you change the acid used, then you change the mobile phase used for the HPLC method. By "replacing" it you are asking if you can change the method. This does not mean you will or will not obtain similar results, just that the method will be changed. Since trichloroacetic acid is also a very strong acid (similar to TFA), you can try it and see. Use the highest quality you can find (HPLC UV grade or equivalent) and at the lowest percentage that results in a reliable and repeatable method.
To find other applicable methods, try a keyword search on the web (e.g. Google).
If the pH is the critical point for mobile phase composition and lipase LC identification, sure TCA may also work for it.
But in the case of protein/enzyme analysis, in general, TFA is added to phases for more than providing acidity but supplying ion pair efficiency. Thereby using TFA as mobile additives can improve the resolution and I do not think TCA may hold the same resolution capability...
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