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Immunoassays for riboflavin and flavin mononucleotide using antibodies specific to D-ribitol and D-ribitol-5-phosphate.

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Relation http://ir.cftri.com/13286/
http://dx.doi.org/10.1016/j.jim.2017.03.010
 
Title Immunoassays for riboflavin and flavin mononucleotide using antibodies specific to
D-ribitol and D-ribitol-5-phosphate.
 
Creator Ravi, G.
Venkatesh, Y. P.
 
Subject 10 Immunology
04 Vitamins
 
Description Riboflavin (vitamin B2), a water-soluble vitamin, plays a key role inmaintaining human health. Though, numerous
methods have been reported for the determination of total riboflavin (TRF) content in foods and biological
samples, very few methods are reported for quantifying riboflavin and its coenzymes [flavin mononucleotide
(FMN); flavin adenine dinucleotide (FAD)] individually. Recently,we have demonstrated that antibodies specific
to D-ribitol and D-ribitol-5-phosphate also recognize riboflavin and FMN, respectively, and not vice-versa. In this
study, we have evaluated these two antibodies for the analysis of riboflavin and FMN by indirect competitive
ELISA (icELISA) in selected foods and pharmaceuticals. Under the optimal assay conditions, 50% inhibition concentration
(IC50) and limit of detection (LOD, IC10) were 3.41 ng/mL and 0.02 ng/mL for riboflavin, and
7.84 ng/mL and 0.24 ng/mL for FMN, respectively, with detectable concentration range between 0.1 and
100 ng of analytes and b0.1% cross-reactivity with otherwater-soluble vitamins. The amounts of TRF in food samples,
as analyzed by icELISA using ribitol antibody, were 90–95% of the reported values in the literature or label
values. Quantification of individual flavins (riboflavin and FMN) from the same food samples showed variation
in their values compared to TRF, and were in good agreement with values obtained from HPLC and AOAC
methods. Further, spiking and recovery analysis of food samples and pharmaceuticals showed no significant matrix
effects. The immunoassays were validated in terms of accuracy and precision using inter- and intra-assays.
The immunoassays developed in this study are sensitive and appears feasible for screening a large number of
samples in the quantification of riboflavin and FMN in various biological samples, pharmaceuticals and natural/
processed foods.
 
Date 2017
 
Type Article
PeerReviewed
 
Format pdf
 
Language en
 
Identifier http://ir.cftri.com/13286/1/Journal%20of%20Immunological%20Methods%20xxx%20%282017%29%20xxx%E2%80%93xxx.pdf
Ravi, G. and Venkatesh, Y. P. (2017) Immunoassays for riboflavin and flavin mononucleotide using antibodies specific to D-ribitol and D-ribitol-5-phosphate. Journal of Immunological Methods, 445. pp. 59-66.