Validated B vitamin quantification from lentils by selected reaction monitoring mass spectrometry

Abstract

A validated method for B vitamin separation and quantification from lentil seeds using ultra high performance liquid chromatography-selected reaction monitoring mass spectrometry (UHPLC-SRM MS) was reported. The use of three enzymes (acid phosphatase, β-glucosidase, and rat serum) with a 4 h incubation was sufficient to convert bound B vitamins into their free forms. Twenty B vitamers were selected and a 5-min UHPLC-SRM MS method was optimized for rapid analysis. This method was applied to quantify B vitamin concentration during lentil seed germination over a 5-day period. Total B vitamins increased up to 1.5-fold on day 5 (from 39.2 µg/g to 60.6 µg/g of dry weight) comparing with dry seeds. Vitamin B5 (pantothenic acid) was the most abundant B vitamin in both dry seeds (34.2%) and in germinated seeds (17.7%-24.5% of total B vitamins); B8 (biotin) and B12 (cyanocobalamin) were not detected in lentil samples.

Keywords: B vitamins; Germination; Lentil; Mass spectrometry; Quantification; Validation.