Quantitation of Fat Soluble Vitamins in Different Walnut Genotypes and Commercial Cultivars; a Comprehensive and Robust LC-MS/MS Method Validation

Walnut, being both nutritious and delicious, has been regarded as a health food due to its preventive and therapeutic effects. In the present study, a robust LC-MS/MS analytical method was developed to simultaneously quantify five fat soluble vitamins (D2, D3, K2, E, K1) in eleven walnut genotypes and two commercial cultivars. Linearity, accuracy, inter- and intra-day precision, detection limit (LOD), quantification limit (LOQ) and uncertainty (U at confidence level of 95% (k = 2)) were the validation parameters of the developed LC-MS/MS method. The established linearity ranges for the studied vitamins were between 50 and 1200 mg L−1 for vitamins D2, D3, K1, between 100-2400 mgL−1 for vitamin E and between 200-4800 mg L−1 for vitamin K2. The coefficients of determination for the vitamins D2, D3, K2, E, K1 were between 0.9990, 0.9991, 0.9995, 0.9997, 0.9991, respectively. Vitamins D3, K2 and K1 were not present in the analysed walnut samples. On the other hand, vitamins E and D2 were in high quantities in all studied walnut samples. D2 and E were the most abundant fat soluble vitamins in MDE32 (29.1 mg g-1 dry plant) and MDE5 (15.2 mg g-1 dry plant) genotypes. Despite the fact that the walnut was selected as real life sample, the validated LC-MS/MS method is enforceable to a broad range of food species.

Quantitation of Fat Soluble Vitamins in Different Walnut Genotypes and Commercial Cultivars; a Comprehensive and Robust LC-MS/MS Method Validation

Walnut, being both nutritious and delicious, has been regarded as a health food due to its preventive and therapeutic effects. In the present study, a robust LC-MS/MS analytical method was developed to simultaneously quantify five fat soluble vitamins (D2, D3, K2, E, K1) in eleven walnut genotypes and two commercial cultivars. Linearity, accuracy, inter- and intra-day precision, detection limit (LOD), quantification limit (LOQ) and uncertainty (U at confidence level of 95% (k = 2)) were the validation parameters of the developed LC-MS/MS method. The established linearity ranges for the studied vitamins were between 50 and 1200 mg L−1 for vitamins D2, D3, K1, between 100-2400 mgL−1 for vitamin E and between 200-4800 mg L−1 for vitamin K2. The coefficients of determination for the vitamins D2, D3, K2, E, K1 were between 0.9990, 0.9991, 0.9995, 0.9997, 0.9991, respectively. Vitamins D3, K2 and K1 were not present in the analysed walnut samples. On the other hand, vitamins E and D2 were in high quantities in all studied walnut samples. D2 and E were the most abundant fat soluble vitamins in MDE32 (29.1 mg g-1 dry plant) and MDE5 (15.2 mg g-1 dry plant) genotypes. Despite the fact that the walnut was selected as real life sample, the validated LC-MS/MS method is enforceable to a broad range of food species.

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  • Abbey M, Noaks M, Belling GB, Nestel PJ, 1994. Partial Replacement of Saturated Fatty Acids with Almonds or Walnuts Lowers Total Plasma Cholesterol and Low-Density-Lipoprotein Cholesterol. American Journal of Clinical Nutrition, 59:995–999.
  • Amaral JS, Alves MR, Seabra RM, Oliveira BPP, 2005. Vitamin E composition of walnuts (Juglans regia L.): a 3-year comparative study of different cultivars. Journal of Agricultural and Food Chemistry, 53: 5467–5472.
  • Awad AB, Fink CS, 2000. Phytosterols as anticancer dietary components: evidence and mechanism of action. Journal of Nutrition, 130: 2127–2130.
  • Bakkalbaşı E, Yılmaz ÖM, Poyrazoğlu ES, Artık N, 2014. Tocopherol contents of walnut varieties grown in Turkey and the Effect of Storage on Tocopherol Content. Journal of Food Processing and Preservation, 38: 518–526.
  • Crews C, Hough P, Godward J, Brereton P, Lees M, Guiet S, Winkelmann W, 2005. Study of the main constituents of some authentic walnut oils. Journal of Agricultural and Food Chemistry, 53: 4853–4860.
  • EURACHEM CITAC Guide CG4. 2004. Quantifiying Uncertainty in Analytical Measurement, 3rd ed.; Ellison, S. L. R., Williams, A.,, Eds.; available from www.eurachem.org (Accessed 5 November 2018).
  • FAO. 2018. Food and Agriculture Organization of the United Nations FAOSTAT, http://www.fao.org/faostat/en/?#data (Accessed 16 October 2018).
  • Fraser GE, Sabaté J, Beeson WL, Strahan TMA, 1992. Possible Protective Effect of Nut Consumption on Risk of Coronary Heart Disease. Archives of Internal Medicine, 152: 1416–1424.
  • Ibarrola-Jurado N, Bulló M, Guasch-Ferré M, Ros E, Martínez-González MA, Corella D, Fiol M, Wärnberg J, Estruch R, Román P, Arós F, Vinyoles E, Serra-Majem L, Pintó X, Covas MI, Basora J, Salas-Salvadó J; PREDIMED Study Investigators, 2013. Cross-sectional assessment of nut consumption and obesity, metabolic syndrome and other cardiometabolic risk factors: the PREDIMED study. PLoS One, 8.
  • Martínez ML, Mattea MA, Maestri DM, 2006. Varietal and crop year effects on lipid composition of walnut (Juglans regia) genotypes. Journal of the American Oil Chemists' Society, 83: 791–796.
  • Rabadán A, Pardo JE, Pardo-Giménez A, Álvarez-Ortí M, 2018. Effect of genotype and crop year on the nutritional value of walnut virgin oil and defatted flour. Science of the Total Environment, 634: 1092-1099.
  • Sabaté J, Fraser GE, Burke K, Knutsen SF, Bennett H, Linstead K.D, 1993. Effects of Walnuts on Serum Lipid Levels and Blood Pressure in Normal Men. The New England Journal of Medicine, 329: 603–607.
  • Santos J, Álvarez-Ortí M, Sena-Moreno E, Rabadán A, Pardo JE, Oliveira BPP, 2017. Effect of roasting conditions on the composition and antioxidant properties of defatted walnut flour. Journal of the Science of Food and Agriculture, 98: 1813–1820.
  • Stuetz W, Schlörmann W, Glei M, 2017. B-vitamins, carotenoids and a-/c-tocopherol in raw and roasted nuts. Food Chemistry, 221; 222–227.
  • Savage GP, Dutta PC, McNeil DL, 1999. Fatty acid and tocopherol contents and oxidative stability of walnut oils. Journal of the American Oil Chemists’ Society, 76: 1059-1063.
  • Tsao R, Yang R, Kramer JKG, Hernandez M, 2007. Fatty Acid Profiles, Tocopherol Contents, and Antioxidant Activities of Heartnut (Juglans ailanthifolia Var. cordiformis) and Persian Walnut (Juglans regia L.). Journal of Agricultural and Food Chemistry, 55 (4): 1164-1169.
  • Yang J, Liu RH, Halim L, 2009. Antioxidant and antiproliferative activities of common edible nut seeds. LWT - Food Science and Technology, 42: 1–8.