Major Phospholipids of Selected Dairy Products as Determined by the HPLC-UVvis and 31P-NMR Methods

Abstract

This study was carried out to determine the major phospholipids in selected dairy products (Beta Serum, Procream and Phospholipids-Rich Dairy products), evaluate the accuracy of the developed fat extraction method for liquid dairy samples and to compare the major phospholipids of different dairy samples obtained using the HPLC-UVvis and 31P-NMR methods. It was found that the developed fat extraction method can be used to estimate the lipid content of liquid dairy samples were still a bit satisfactory. Using the HPLC-UVvis method, it was found that the sphingomyelin (SM) consists of 2 curves in Beta Serum, Procream and Phospholipid-Rich products. The phosphatidylethanolamine (PE) separated ahead of phosphatidylcholine (PC) and then PC ahead of SM1 and SM2. The results showed that the data of the major phospholipids (PC, PE and SM) in Butter Serum, Procream and Phospholipids-Rich products as determined by the HPLC-UVvis method compared well with those of the 31P-NMR method It must be noted that the analysis temperature for the HPLC-UVvis method was at 40oC while the 31P-NMR method was at 30oC, and this might have also contributed to the slight variation of the results. Furthermore, the HPLC-UVvis method is rapid and with cheaper analysis cost compared with the 31P-NMR method.

Keywords

• Phospholipids
• Phosphatidylcholine
• Phosphatidylethanolamine
• Sphingomyelin
• HPLC-UVvis
• 31P-NMR

References

1. Bligh EG, Dyer WJ 1959. A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology. 37: 911-917.
2. Contarini G, Povolo M 2013. Phospholipids in milk: composition, biological and technological significance, and analytical strategies. International Journal of Molecular Sciences. 14: 2808-2831.
3. Castro-Gomez P, Rodriguez-Alcala LM, Monteiro KM, Ruiz AL, Carvalho JE, Fontecha J 2016. Antiproliferative activity of butter milk lipid fractions isolated using food grade and non-food grade solvents on human cancer cell lines. Food Chemistry. 212: 695-702.
4. Dewettinck, K, Rombaut R, Thienpont N, Le TT, Messens K, van Camp J 2008. Nutritional and technological aspects of milk fat globule membrane material. International Dairy Journal. 18: 436-457.
5. Donato P, Cacciola F, Chichello F, Russo M, Dugo P, Mondello L 2011. Determination of phospholipids in milk samples by means of hydrophilic interaction liquid chromatography coupled to evaporative light scattering and mass spectrometry detection. Journal of Chromatography.1218: 6476-6482.
6. Duivenvoorden I, Voshol PJ, Rensen PC, van Duyvenvoorde W, Romijin JA, Emeis JJ, Havekes LM, Nieuwenhuizen W.F.. 2006. Dietary sphinolipids lower plasma cholesterol and triacylglycerol and prevent liver steatosis in APOE*3Leiden mice. American Journal of Clinical Nutrition. 84: 312-321.
7. Ferreiro T, Gayoso L, Rodriguez-Otero JL 2017. Determination of phospholipids in milk by HPLC with evaporative light scattering detector: Optimization and validation. Journal of Dairy & Veterinary Sciences. 1 (3): 555562. doi: 10.19080/JDVS.2017.01.555562.
8. Folch JM, Lees M, Sloane-Stanley GH 1957. A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biology and Chemistry. 226: 497-509.

9. Kielbowicz G, Micke P, Wawrzenczyk C 2013. A new liquid chromatography method with charge aerosol detector (CAD) for the determination of phospholipid classes. Application to Milk Phospholipids. Talanta.105: 28-33.
10. Kivinen A, Tarpila S, Salminen S, Vapaatalo H 1992. Gastroprotection with milk phospholipids: a first human study. Milchwissenschaft. 47: 694-696.
11. Kuchta AM, Kelly PM, Stanton C, Devery RA 2012. Milk fat globule membrane – a source of polar lipids for colon health? – A review. International Journal of Dairy Technology. 65: 315-333.
12. Le TT, Miocinovic J, Nguyen TM, Rombaut R, van Camp J, Dewettinck K 2011. Improved solvent extraction procedure and high-performance liquid chromatography-evaporative light scattering detector method for analysis of polar lipids from dairy materials. Journal of Agricultural and Food Chemistry. 59: 10407-10413.
13. Lee CM, Trevino B, Chaiyawat M 1996. A simple and rapid solvent extraction method for determining total lipids in fish tissue. Journal of AOAC International. 79: 487-492.
14. Lesser ME, Sagalowicz L, Michel M, Watzke HJ 2006. Self-assembly of polar food lipids. Advances in Colloid Interface Science. 123: 125-136.
15. Mackenzie A, Vussotski M., Nekrasov E 2009. Quantitative analysis of dairy phospholipids by 31P NMR. Journal of the American Oil Chemists Society. 86: 757-763.
16. McDaniel M, Maier S, Einstein G 2003. Brain-specific nutrients: a memory cure? Nutrition. 19: 957-975.
17. Rehman S, Rehman S, Welter D, Wildenauer D, Ackenheil M 2017. Simple and rapid separation and determination of phospholipids by the HPLC-UV system. Annals of Pharmacology and Pharmaceutics. 2: 1-3.
18. Rombaut R, Dewettinck K 2006. Properties, analysis and purification of milk polar lipids. International Dairy Journal. 16: 1362-1373.
19. Rombaut R, Camp JV, Dewettinck K 2005. Analysis of phospho- and sphingolipids in dairy products by a new HPLC method. Journal of Dairy Science. 88: 482-488.
20. Tetra Pak. 2005. Dairy Processing Handbook. Tetra Pak Processing Systems AB, Lund, Sweden. 482 pp.
21. Vanhoutte B, Rombaut R, Dewettinck K, van der Meeren P 2004. Phospholipids. In Food Analysis, Volume 1. (M. L. Nollet, ed.), Marcel Dekker, New York, New York. pp. 349-382.
22. Verardo V, Gomez-Caravaca AM, Arraez-Roman D, Hettinga K 2017. Recent advances in phospholipids from colostrum, milk and dairy by products. International Journal of Molecular Science. 18: 173. doi: 10.3390/ijms18010173.
23. Vesper H, Schmelz E, Nikolova-Karakashian MN, Dillehay DL, Lynch DV, Merril AH Jr. 1999. Sphingolipids in food and the emerging importance of sphingolipids to nutrition. Journal of Nutrition. 129: 1239-1250.