In conclusion, this study demonstrated that microplate readers
will underestimate the sensitivity for colorimetric analysis compared
to data from a 1-cm pathlength spectrophotometer. However,
the effective optical pathlength for a microplate reader can
be readily determined under conditions not far removed those
used for the mFRAP assay. The molar absorptivity values for gallic
acid and ascorbic acid were determined clearly for the first time
using the mFRAP format. Using the average calibration parameters
for mFRAP1 and 2, the minimum detectable concentration and
upper limit of linearity were determined (Owusu-Apenten, 2002)
as 0.92 107 and 250 107 M for gallic acid, respectively. For
ascorbic acid the minimum detectable concentration and upper
limit of linearity was 2.0 107 and P670 107 M, respectively.
Analysis of New Zealand Manuka honey showed that the total antioxidant
capacity is related to the UMF rating. The pathlength corrections
described here should be applicable to other microplate
based assays for total antioxidant capacity. The methodology
detailed in the current could be useful in evaluating antioxidant
assays on a variety of different platforms.
Acknow
In conclusion, this study demonstrated that microplate readerswill underestimate the sensitivity for colorimetric analysis comparedto data from a 1-cm pathlength spectrophotometer. However,the effective optical pathlength for a microplate reader canbe readily determined under conditions not far removed thoseused for the mFRAP assay. The molar absorptivity values for gallicacid and ascorbic acid were determined clearly for the first timeusing the mFRAP format. Using the average calibration parametersfor mFRAP1 and 2, the minimum detectable concentration andupper limit of linearity were determined (Owusu-Apenten, 2002)as 0.92 107 and 250 107 M for gallic acid, respectively. Forascorbic acid the minimum detectable concentration and upperlimit of linearity was 2.0 107 and P670 107 M, respectively.Analysis of New Zealand Manuka honey showed that the total antioxidantcapacity is related to the UMF rating. The pathlength correctionsdescribed here should be applicable to other microplatebased assays for total antioxidant capacity. The methodologydetailed in the current could be useful in evaluating antioxidantassays on a variety of different platforms.Acknow
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