Glycoprofiling of Oligosaccharides of Regular and Lactose-Free Milk by Mass Spectrometry

Abstract

Oligosaccharides from regular milk and lactose-free milk were analyzed by Electrospray Ionization interface coupled with Ion Trap Mass Spectrometry (IT-MS). The negative mode mass spectrometry of sugar compositions was obtained either by direct infusion and mass spectrometry with Liquid Chromatography (LC). Hexose (Hex) and the other oligomeric sugar components were observed in both regular and lactose-free milk. While lactose derivatives (deprotonated lactose dimer, chloride adducts, chloride dimer) in regular milk have seen to be dominated, monosaccharide derivatives (Hex-H2O, Hex, Hex-Cl) in the lactose-free milk were observed as abundant which was lytic product of lactose. Phosphate/sulfate ester substitution on lactose and sialyllactose in both regular and lactose-free milk samples were observed with similar intensity thus it has been understood that these important milk components are not digested during lactose removing process.

Keywords

• Milk
• Lactose-Free
• LC-MS
• Oligosaccharides
• Sialic acid

References

1. [1]. Costa, A, Lopez-Villalobos, N, Sneddon, NW, Shalloo, L, Franzoi, M, De Marchi, M, Penasa, M. 2019. Invited review: Milk Lactose-Current Status and Future Challenges in Dairy Cattle. Journal of Dairy Science; 102: 5883–5898.
2. [2]. Barłowska, J, Szwajkowska, M, Litwińczuk, Z, Król, J. 2011. Nutritional Value and Technological Suitability of Milk from Various Animal Species Used for Dairy Production. Comprehensive Reviews in Food Science and Food Safety; 10: 291–302.
3. [3]. Adam, AC, Rubio-Texeira, M, Polaina, J. 2004. Lactose: The milk sugar from a biotechnological perspective. Critical Reviews in Food Science and Nutrition; 44: 553–557.
4. [4]. Urashima, T, Saito, T, Nakamura, T, Messer, M. 2001. Oligosaccharides of milk and colostrum in non-human mammals. Glycoconjugate Journal; 185: 357–371.
5. [5]. Swagerty, DL, Walling, AD, Klein, RM. 2002. Lactose Intolerance. American Academy of Family Physicians; 65: 1855–1861.
6. [6]. Sharp, E, D’Cunha, NM, Ranadheera, CS, Vasiljevic, T, Panagiotakos, DB, Naumovski, N. 2021. Effects of lactose-free and low-lactose dairy on symptoms of gastrointestinal health: A systematic review. International Dairy Journal; 114: 104936.
7. [7]. Harju, M, Patent. 1989. Process for the specific separation of lactose from milk. US Patent 4820348A.
8. [8]. Harju, M, Kallioinen, H, Tossavainen, O. 2012. Lactose hydrolysis and other conversions in dairy products: Technological aspects. International Dairy Journal; 22(2): 104-109.

9. [9]. Dekker, PJT, Koenders, D, Bruins, MJ. 2019 .Lactose-Free Dairy Products: Market Developments, Production, Nutrition and Health Benefits. Nutrients; 11(3): 551.
10. [10]. Tao, N, DePeters, EJ, Freeman, S, German, JB, Grimm, R, Lebrilla, CB. 2008. Bovine milk glycome. Journal of Dairy Science. 91(10): 3768–3778.
11. [11]. Açar, Y, Yassibaş, E. 2021. Anne Sütü Oligosakkaritleri ve Sağlık Üzerine Etkileri. Gazi Sağlık Bilimleri Dergisi. 6: 22–33.
12. [12]. Bode, L. 2015. The functional biology of human milk oligosaccharides. Early Human Development. 91: 619–22.
13. [13]. Wang, B, Yu, B, Karim, M, Hu, H, Sun, Y, McGreevy, P, Petocz, P, Held, S, Brand-Miller, J. 2007. Dietary sialic acid supplementation improves learning and memory in piglets. American Journal of Clinical Nutrition. 85(2): 561–569.
14. [14]. Tarr, AJ, Galley, JD, Fisher, SE, Chichlowski, M, Berg, BM, Bailey, MT. 2015. The prebiotics 3′Sialyllactose and 6′Sialyllactose diminish stressor-induced anxiety-like behavior and colonic microbiota alterations: Evidence for effects on the gut–brain axis. Brain, Behavior, and Immunity. 50: 166–177.
15. [15]. Mudd, AT, Salcedo, J, Alexander, LS, Johnson, SK, Getty, CM, Chichlowski, M, Berg, BM, Barile, D, Dilger, RN. 2016. Porcine Milk Oligosaccharides and Sialic Acid Concentrations Vary Throughout Lactation. Frontiers in Nutrition. 3: 39.
16. [16]. Tao, N, DePeters, EJ, German, JB, Grimm, R, Lebrilla, CB. 2009. Variations in bovine milk oligosaccharides during early and middle lactation stages analyzed by high-performance liquid chromatography-chip/mass spectrometry. Journal of Dairy Science. 92(7): 2991–3001.
17. [17]. Ninonuevo, MR, Park, Y, Yin, H, Zhang, J, Ward, RE, Clowers, BH, German, JB, Freeman, SL, Killeen, K, Grimm, R, Lebrilla, CB. 2006. A strategy for annotating the human milk glycome. Journal of Agricultural and Food Chemistry. 54(20): 7471–7480.
18. [18]. Kirmiz, N, Robinson, RC, Shah, IM, Barile, D, Mills, DA. 2018. Milk Glycans and Their Interaction with the Infant-Gut Microbiota. Annual Review of Food Science and Technology. 9: 429-450.
19. [19]. Kelly, V, Davis, S, Berry, S, Melis, J, Spelman, R, Snell, R, Lehnert, K, Palmer, D. 2013. Rapid, quantitative analysis of 3′- and 6′-sialyllactose in milk by flow-injection analysis–mass spectrometry: Screening of milks for naturally elevated sialyllactose concentration. Journal of Dairy Science. 96: 7684–7691.
20. [20]. Remoroza, CA, Mak, TD, Lorna, M, De Leoz, A, Mirokhin, YA, Stein, SE. 2018. Creating a Mass Spectral Reference Library for Oligosaccharides in Human Milk. Analytical Chemistry. 90(15): 8977-8988.
21. [21]. Mathon, C, Barding, GA, Larive, CK. 2017. Separation of ten phosphorylated mono-and disaccharides using HILIC and ion-pairing interactions. Analytica Chimica Acta. 972: 102–110.
22. [22]. Panseri, S, Pavlovic, R, Castrica, M, Nobile, M, Di Cesare, F, Chiesa, LM. 2021. Determination of carbohydrates in lactose-free dairy products to support food labelling. Foods. 10(6): 1219.
23. [23]. Ilves, A, Harzia, H, Ling, K, Ots, M, Soomets, U, Kilk, K. 2012. Alterations in milk and blood metabolomes during the first months of lactation in dairy cows. Journal of Dairy Science. 95(10): 5788–5797.
24. [24]. Li, J, Jiang, M, Zhou, JR, Ding, J, Guo, Z, Li, M, Ding, F, Chai, W, Yan, J, Liang, X. 2021. Characterization of rat and mouse acidic milk oligosaccharides based on hydrophilic interaction chromatography coupled with electrospray tandem mass spectrometry. Carbohydrate Polymers. 259: 117734.
25. [25]. Cai, Y, Cole, RB. 2002. Stabilization of anionic adducts in negative ion electrospray mass spectrometry. Analytical Chemistry. 74(5): 985–991.
26. [26]. Guan, B, Cole, RB. 2008. MALDI Linear-Field Reflectron TOF Post-Source Decay Analysis of Underivatized Oligosaccharides: Determination of Glycosidic Linkages and Anomeric Configurations Using Anion Attachment. Journal of the American Society for Mass Spectrometry. 19(8): 1119–1131.
27. [27]. Madson, MA. Mass Spectrometry: Techniques for Structural Characterization of Glycans, 1st edn. Elsevier Press: USA, 2016, pp 1-17.
28. [28]. Araújo, AS, Da Rocha, LL, Tomazela, DM, Sawaya, ACHF, Almeida, RR, Catharino, RR, Eberlin, MN. 2005. Electrospray ionization mass spectrometry fingerprinting of beer. Analyst. 130(6): 884–889.
29. [29]. Moriwaki, H, Hagiwara, A, Takasaki, M, Izumi, F, Watanabe, A, Shimizu, R, Kuribayashi, N, Totani, Y, Suzuki, Y. 2010. Electrospray ionization-mass spectrometric measurement of sake, a traditional japanese alcohol beverage, for characterization. Analytical Sciences. 26(3): 379-382.
30. [30]. Petzold, CJ, Leavell, MD, Leary, JA. 2004. Screening and Identification of Acidic Carbohydrates in Bovine Colostrum by Using Ion/Molecule Reactions and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry: Specificity toward Phosphorylated Complexes. Analytical Chemistry. 76(1): 203–210.
31. [31]. Fong, B, Ma, K, McJarrow, P. 2011. Quantification of bovine milk oligosaccharides using liquid chromatography-selected reaction monitoring-mass spectrometry. Journal of Agricultural and Food Chemistry. 59(18): 9788–9795.
32. [32]. Wheeler, SF, Harvey, DJ. 2000. Negative ion mass spectrometry of sialylated carbohydrates: Discrimination of N-acetylneuraminic acid linkages by MALDI-TOF and ESI-TOF mass spectrometry. Analytical Chemistry. 72(20): 5027–5039.
33. [33]. Chai, W, Piskarev, VE, Mulloy, B, Liu, V, Evans, PG, Osborn, HMI, Lawson, AM. 2006. Analysis of chain and blood group type and branching pattern of sialylated oligosaccharides by negative ion electrospray tandem mass spectrometry. Analytical Chemistry. 78(5): 1581–1592.