The study was carried out to develop a method for determining the biotin content in nutritional products and supplements by liquid chromatography-mass spectrometry (LC-MS/ MS) combined with a fast and simple pretreatment. The analyzed sample was hydrolyzed in acetate buffer (50 mM, pH 4) with the presence of ascorbate solution (10%) at 120 ± 1°C. After adding the internal standard (2H4-biotin), the mixture was analyzed on LC-MS/ MS system using C18 (100 mm × 2.1 mm; 1.7 µm) column and 0.1% formic acid and MeOH as mobile phases. The analyte was detected on an MS/MS system with an ESI (+) ionization source. The method was validated following the AOAC criteria. The method detection limit was 0.15 - 4.20 µg/100g, the method quantitative limit was in the range of 0.5 - 14.0 µg/100g, the recovery range was 81.3 - 107.3%, the repeatability was in the range of 2.00 - 7.30 %, the reproducibility was 2.50 - 7.70%, which were in accordance with AOAC requirements. The method has been applied to analyze biotin content in 10 nutritional products and 10 supplements purchased in the market.
Biotin, LC-MS/MS, nutritional products, supplements.
[1]. Höller. U, Wachter. F, Wehrli. C, Fizet. C, “Quantification of biotin in feed, food, tablets, and premixes using HPLC-MS/MS,” Journal of Chromatography B, vol. 831, no. 1-2, pp. 8-16, 2006.
[2]. B C. Nelson, Katherine E. Sharpless, Lane C. Sander, “Improved Liquid Chromatography Methods for the Separation and Quantification of Biotin in NIST Standard Reference Material 3280: Multivitamin/Multielement Tablets,” Journal of Agriculture and Food Chemistry, vol. 54, no. 23, pp. 8710-8716, 2006.
[3]. TCVN 9044:2012. Determination of d-biotin in foods by liquid chromatography.
[4]. Fan. B, You. J, Suo. Y, Qian. C, “A novel and sensitive method for determining vitamin B3 and B7 by pre-column derivatization and high-performance liquid chromatography method with fluorescence detection,” Plos One, vol. 13, no.6, pp. 1-15, 2018.
[5]. Kakitani. A, Inoue. T, Matsumoto. K, Watanabe. J, Nagatomi. Y, Mochizuki. N, “Simultaneous determination of water-soluble vitamins in beverages and dietary supplements by LC-MS/MS,” Food Additives & Contaminants: Part A, vol. 31, no. 12, pp. 1939-1948, 2014.
[6]. B. D. Gill, S. Saldo, J. E. Wood, H. E. Indyk, “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,” Journal of AOAC International, vol. 101, mo. 5, pp. 1578-1583, 2018.
[7]. Nierves. L, Lange. P. F, “Detectability of Biotin Tags by LC-MS/MS,” Journal of Proteome Research, vol. 20, no. 5, pp. 3002-3008, 2021.
[8]. AOAC Official Method 2016.02. Total Biotin in Infant Formula and Adult/Pediatric Nutritional Formulas Liquid Chromatography Coupled with Immunoaffinity Column Cleanup Extraction.
[9]. T. C. Son, "Method validation and measurement uncertainty assessment in chemical analysis,” Science and Technics Publishing House, 2021.