Good day everyone!
I am currently working on measuring ATP content from drinking water samples. Our team´s main goal by now is to achieve the best luminescent signal by disrupting as many cells as possible. We´re interested in using TCA 5% as lysis reagent, however, we can´t find a proper protocol or technique to neutralize it, specially the concentrations needed to do this. Also, our sample volume is very small (around 1-2 ml) hence the handling is a bit complicated. We´ve read articles mentioning NaOH, acetone or other alkalis, but they don´t mention concentrations of the reagents or used to neutralize the TCA.
I hope you can help us with this issue. Thank you very much in advance for your help!
Try removing TCA by repeated extraction with distilled diethyl ether. Then residual ether can be removed under stream of nitrogen. Try this . Hopefully this will work.
Good Luck!
Hi, The best way to remove TCA is the method described by Dr. Katyare. The method is described in the attached paper from 1975. In this paper I used EDTA, which was stupid as EDTA does not complex metal ions at acid pH. The ether should be water saturated as it will otherwise concentrate the water phase. TCA is fine as a reference method, but not very practical if TCA must be removed (pH neutralization is not efficient as the trichloroacetate ion is inhibitory in itself. Quaternary ammonium compounds is a better alternative (cf. 1984 paper) as they can be inactivated by cyclodextrins (cf. 1994 paper). Kits based on this principle are available from BioThema. One of these kits also includes degradation of extracellular ATP, which is often high in drinking water. Please contact me by email and I can tell you all about it. Best regards, Arne
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