Brendon Gill PhD MSc(hons) GradDipT FAOACI


A Rapid Method for the Determination of Biotin and Folic Acid in Liquid Milk, Milk Powders, Infant Formula, and Milk-based Nutritional Products by Liquid Chromatography-Tandem Mass Spectrometry

Brendon D. Gill, Sheila Saldo, Jackie E. Wood, Harvey E. Indyk

Background: Biotin and folate are B-group vitamins that play a critical role in numerous metabolic reactions and are supplemented to infant and adult nutritional formulas as free biotin and folic acid. Objective: A rapid method for the analysis of biotin and folic acid that is applicable to liquid milk, milk powders, infant formula, and milk-based nutritional products is described. Methods: Samples are autoclaved, centrifuged, filtered, and analyzed by high performance liquid chromatography-tandem mass spectrometry, with quantitation accomplished by the internal standard technique. Results: The method was shown to be accurate, with acceptable spike recovery (biotin: 96.5–108.2%; folic acid: 92.6–104.4%), and no bias (α = 0.05) against either a certified reference material (biotin: p = 0.70; folic acid: p = 0.23) or established analytical methods (biotin: p = 0.10; folic acid: p = 0.48) was found. Acceptable precision was confirmed with repeatability relative standard deviation and Horwitz ratio values (biotin: RSDr = 0.5–5.6%, HorRatr = 0.1–0.6; folic acid: RSDr = 2.0–3.1%, HorRatr = 0.3–0.5). Method detection limit and ruggedness experiments further demonstrated the suitability of this method for routine compliance testing. Conclusions: This rapid method is intended for use in high-throughput laboratories as part of routine product compliance release testing of biotin and folic acid as part of the manufacture of infant formulas and adult nutritional products.



Gill et al. (2018) J. AOAC Int., 101, 1578–1583.Journal Link
Gill et al. (2018) J. AOAC Int., 101, 1578–1583.Article PDF
Reference Style

Journal: Journal of AOAC International
Publisher: AOAC International

Gill, B.D., Saldo, S., Wood, J.E., & Indyk, H.E. (2018) A Rapid Method for the Determination of Biotin and Folic Acid in Liquid Milk, Milk Powders, Infant Formula, and Milk-based Nutritional Products by Liquid Chromatography-Tandem Mass Spectrometry. J. AOAC Int. 101, 1578‑1583
doi: 10.5740/jaoacint.18-0065


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