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Determination of folate/folic acid level in milk by microbiological assay, immuno assay and high performance liquid chromatography

Published online by Cambridge University Press:  18 March 2013

Ramya Iyer  and
Sudhir Kumar Tomar
Ramya Iyer
Affiliation:
Dairy Microbiology Division, National Dairy Research Institute, Karnal, India
Sudhir Kumar Tomar*
Affiliation:
Dairy Microbiology Division, National Dairy Research Institute, Karnal, India
*
*For correspondence; e-mail: sudhirndri@gmail.com
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Abstract

A concurrent determination of folate versus folic acid in milk by microbiological assay (MA) with Lactobacillus rhamnosus as the assay organism, Enzyme Linked Immuno Sorbent Assay (ELISA) by competitive binding rapid ELISA kit (RIDASCREEN®) and high-pressure-liquid chromatography (HPLC) was done for detection of the folate form and its level. MA gave total folate content as Lb. rhamnosus showed similar response to most folate isomers formed by the tri-enzyme treatment in comparison with the other two methods which specifically estimated the folic acid. In case of ELISA, specificity was apparently limited to folic acid and dihydro folic acid and thereby showed a lower response for other folate derivatives. Estimation by HPLC with UV detector was highly specific and hence only folic acid could be detected without any cross reactivity. Among the different methods HPLC was observed to be the most sensitive method for determination of folic acid and hence can efficiently determine the folic acid fortification level while MA remained highly efficient, sensitive and reproducible method for estimation of total folate indicating its potential use for dietary folate estimation.

Keywords

Folic acidmilkfortificationmicrobiological assayenzyme linked immuno sorbent assayhigh performance liquid chromatography
Type
Research Article
Information
Journal of Dairy Research , Volume 80 , Issue 2 , May 2013 , pp. 233 - 239
Copyright
Copyright © Proprietors of Journal of Dairy Research 2013

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