Analytical and Bioanalytical Chemistry

, Volume 409, Issue 27, pp 6475–6484 | Cite as

Assessment of bioavailable B vitamin content in food using in vitro digestibility assay and LC-MS SIDA

  • Toomas Paalme
  • Allan Vilbaste
  • Kaspar Kevvai
  • Ildar Nisamedtinov
  • Kristel Hälvin-Tanilas
Research Paper


Standardized analytical methods, where each B vitamin is extracted from a given sample individually using separate procedures, typically ensure that the extraction conditions provide the maximum recovery of each vitamin. However, in the human gastrointestinal tract (GIT), the extraction conditions are the same for all vitamins. Here, we present an analytically feasible extraction protocol that simulates conditions in the GIT and provides a measure of the content of bioavailable vitamins using LC-MS stable isotope dilution assay. The results show that the activities of both human gastric and duodenal juices were insufficient to liberate absorbable vitamers (AV) from pure cofactors. The use of an intestinal brush border membrane (IBBM) fraction derived from the mucosal tissue of porcine small intestine ensured at least 70% AV recovery. The rate of AV liberation, however, was strongly dependent on the cofactor, e.g., in the case of NADH, it was magnitudes higher than in the case of thiamine diphosphate. For some vitamins in some food matrices, the use of the IBBM fraction assay resulted in lower values for the content of AV than conventional vitamin determination methods. Conventional methods likely overestimate the actual bioavailability of some vitamins in these cases.

Graphical abstract

Assessment of bioavailable B vitamin content in food


B vitamins Bioavailability Absorbable vitamers LC-MS Stable isotope dilution assay 



Financial support for this research was provided by the European Regional Development Fund (project EU48667) and Estonian Ministry of Education (institutional research funding IUT19-27).

Compliance with ethical standards

Human and animal rights

This article does not contain any studies with animals performed by any of the authors.

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Toomas Paalme
    • 1
  • Allan Vilbaste
    • 2
  • Kaspar Kevvai
    • 2
  • Ildar Nisamedtinov
    • 1
    • 2
    • 3
  • Kristel Hälvin-Tanilas
    • 2
  1. 1.Department of Chemistry and BiotechnologyTallinn University of TechnologyTallinnEstonia
  2. 2.Center of Food and Fermentation TechnologiesTallinnEstonia
  3. 3.Lallemand, Inc.MontrealCanada

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