The method of simultaneous quantification some of Human Milk Oligosaccharide in dietary supplements by LC-MS/MS is an accurate and effective method to quickly determine the content of 2'-Fucosyllactose (2 '-FL), Lacto-N-neotetraose (LNnT), Lacto-N-tetraose (LNT), 3'-Siallylactose (3'-SL) and 6'-Siallylactose (6'-SL) in both dietary supplements powder and liquid. The method has been developed and validated follow the AOAC International guidelines. The mobile phase system consists of 2 channels: channel A (0.1% formic acid) and channel B (acetonitril) connected to the HILIC column (3.5 μm, 2.1mm × 150 mm) and the MS detector. The time analysis is 10 minutes, this method can identify all 5 substances belonging to the group HMOs. The detection limit and quantitative limit for all 5’-FL, LNnT, LNT, 3’-SL, 6’-SL were 4 mg/kg and 10 mg/kg, respectively. The linear range of the method ranges from 0.4 µg/mL to 40 µg/mL. Other validation parameters include the accuracy (R% 98.8 -103%); The precision (RSD% 1.69 - 5.54%) can meet the requirement of AOAC. The method was applied in practice to analyze 25 supplementary food samples on the market gives the results of analyzing total HMOs in powdered samples about 0.1 – 0.3 g/100g and for liquid samples about 0.01 – 0.03 g/100mL.
2’-Fucosyllactose (2’-FL), lacto-N-tetraose (LNT), HMOs; dietary supplement, LC-MS/MS
[1]. American pregnance association, “Breastfeeding Overview”. [Online]. Availble: https://americanpregnancy.org/healthy-pregnancy/breastfeeding/breastfeeding-overview-copy-70904/ [Aceessed 02/19/2020]
[2]. M. H. Monaco, J. Kim, and S. M. Donovan, “Human Milk: Composition and Nutritional Value,” Encyclopedia of Food and Health, vol. 2016, pp. 357-362, 2016.
[3]. M. Li, “Human milk oligosaccharides shorten rotavirus-induced diarrhea and modulate piglet mucosal immunity and colonic microbiota,” ISME Journal, vol. 8, no. 8, pp. 1609-1620, 2004.
[4]. A. L. Morrow, G. M. Ruiz-Palacios, X. Jiang, and D. S. Newburg, “Human-milk glycans that inhibit pathogen binding protect breast-feeding infants against infectious diarrhea,” Journal of Nutrition, vol. 135, no. 5, pp. 1304-130, 2005.
[5]. S. S. Leeuwen, “Challenges and Pitfalls in Human Milk Oligosaccharide Analysis,” Nutrients, vol. 11, no. 11, pp. 2684, 2019.
[6]. A. S. Christensen, S. H. Skov, S. E. Lendal, and B. H. Hornshoj, “Quantifying the human milk oligosaccharides 2’‐fucosyllactose and 3‐fucosyllactose in different food applications by high‐performance liquid chromatography with refractive index detection,” Journal of Food Science, vol. 85, no. 2, pp. 332-339, 2010.
[7]. S. Austin, D. Cuany, J. Michaud, B. Diehl, and Begoña Casado, “Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant Formula”, Molecules, vol. 23, no. 10, pp. 2650, 2010.
[8]. S. S. van Leeuwen, “Challenges and Pitfalls in Human Milk Oligosaccharide Analysis”, Nutrients, vol. 11, no. 11, pp. 2684, 2019.
[9]. T. R. I. Cataldi, C. Campa, and G. E. De Benedetto, “Carbohydrate analysis by high-performance anion-exchange chromatography with pulsed amperometric detection: The potential is still growing”, Analytical and Bioanalytical Chemistry, vol. 368, no. 8, pp. 739–758, 2000.
[10]. Y. Bao, C. Chen, and D. S. Newburg, “Quantification of neutral human milk oligosaccharides by graphitic carbon high-performance liquid chromatography with tandem mass spectrometry”, Analytical Biochemistry, vol. 433, no. 1, pp. 28–35, 2013.