I have performed an alpha-amylase inhibition test by Acarbose (commercially available tablet). My indicator is DNSA. Normally higher the acarbose concentration, more alpha-amylase inhibited, hence less glucose produce by starch degradation and ultimately color intensity and absorption values of the sample (by reducing DNSA with glucose) is low.
In contrast, I observed opposite results repeatedly for several times. That means the highest color observed in highest acarbose containing the sample.
Did anyone have this experience?
What could be the reason for that?
Is acarbose act as reducing sugar and interfere with the test results?
How can I overcome this problem?
To find out whether the acarbose is causing interference in the assay, perform the same reaction again, this time leaving out the amylase.
Thank you Dr. Adam B Shapiro for your suggestion. I will do so.
Try using heat inactivated amylase without putting acarbose keeping reaction system intact. This will let you know the effect of acarbose.
Thanks for your suggestion Dr. Shamsher Singh Kanwar . We used the amylase at refrigerated conditions. We did all the negative and positive controls e.g. without amylase, without acarbose, etc. It seems like acarbose tablet reacts with the DNS, the color agent.
We the research group wonder how this happened and why other scientists did not detect this before. We used acarbose tablets made in India and Germany.
We have found that the color of test sample (e.g. plant extract) is interfering with the results. Therefore, with each test sample a positive control should be tested to mitigate the effect of sample color.
Yes, the extract interfere with the analysis of glucose by DNS method
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