blank for starch iodine method

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blank for starch iodine method

Postby apusi » Aug 15 2018 7:24 pm

what should I use as blank in starch iodine method for detecting antidiabetic activity or alpha amylase inhibitory effect
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Re: blank for starch iodine method

Postby mdfenko » Aug 16 2018 9:30 pm

if for no other reason, to zero your spectrophotometer
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Re: blank for starch iodine method

Postby apusi » Aug 17 2018 5:27 pm

i know its to zero the spectrophotometer. i am getting negative value .so i want to know what is included in blank solution for starch iodine method to check antidiabetic activity.
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Re: blank for starch iodine method

Postby mdfenko » Aug 17 2018 9:53 pm

oops, sorry, i read "why" where it said "what".

normally, the blank would be everything in the assay but without starch (or the potential source of starch) added (replaced by water or the buffer in which the source is solubilized). if that still leads to a negative reading, then try with the source but not the iodine.
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Re: blank for starch iodine method

Postby apusi » Aug 18 2018 2:24 am

thanks for replying. previously I used reagent blank which included DMSO +phosphate buffer solution (since extract was dissolved in DMSO and then furthur diluted with phosphate buffer), phosphate buffer (since alpha amylase was dissolved in phosphate buffer), distilled water (since starch was dissolved in distilled water), hcl and Iodine. i haven't included extract, alpha amylase, starch in blank because they were analytes. i just used reagent blank to zero. is this the right way ??
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Re: blank for starch iodine method

Postby mdfenko » Aug 20 2018 9:19 pm

it appears to be okay, although i may have added starch to the assay (complete assay without the enzyme).

one other way would be to prepare a complete assay and blank at zero time. then take readings at set time points or, if you have a chart recorder, follow the change in absorbance over time.
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