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Pantothenic acid (vitamin B5) in fortified foods: comparison of a novel ultra-performance liquid chromatography-tandem mass spectrometry method and a microbiological assay (AOAC Official Method 992.07).


A novel method was developed and single-laboratory validated for the determination of free pantothenic acid (vitamin B5) in a wide range of infant and adult fortified food products. The method combines simple sample preparation and chromatographic analysis using ultra-performance LC coupled to tandem MS with positive electrospray ionization. Pantothenic acid was quantified using [13C6, 15N2]-pantothenic acid as an internal standard. Calibration curves were linear between 0.08 and 1.2 microg/mL (r2 = 0.9998), and average recovery varied between 95 and 106%. The method exhibited overall RSD(r) of 1.1% and RSD intermediate reproducibility from 2.5 to 6.0% in infant formulas and cereals. Comparison of results between total and free pantothenic acid showed that the analysis of free pantothenic acid gave a good estimation of total pantothenic acid in the range of products analyzed. The method provides reliable free pantothenic acid results in a wide range of fortified foods (infant and adult nutritionals, cereal products and beverages), and shows good correlation with the microbiological method AOAC Official Method 992.07. It is a more selective, faster, and robust alternative to microbiological determination.


  • Publisher Full Text
  • Authors+Show Affiliations


    Nestlé Research Center, Vers-chez-les-blanc, CH-1000 Lausanne 26, Switzerland.

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    Biological Assay
    Chromatography, High Pressure Liquid
    Food, Fortified
    Indicators and Reagents
    Infant Food
    Lactobacillus plantarum
    Pantothenic Acid
    Reference Standards
    Regression Analysis
    Reproducibility of Results
    Tandem Mass Spectrometry

    Pub Type(s)

    Comparative Study
    Journal Article



    PubMed ID