A novel application of the liquid chromatography method combined with the triple quadrupole tandem mass spectrometry method was developed for the quantification of vitamin K1 and two forms of vitamin K2 (menaquinone-4, menaquinone-7) in human serum. Total chromatography time for each run was 9 min. Time required for the sample pretreatment procedures was approximately 4 h. The coefficients of variation (CVs) of intra-assay were 10.4%, 3.2 % and 2.3% for vitamin K1 in three levels of quality control samples; were 14.3%, 3.2% and 6.7% for menaquinone-4; and were 11.1%, 6.0% and 7.0% for menaquinone-7. The inter-assay CVs were 12.8%, 11.3% and 7.4% for vitamin K1; were 15.2%, 9.2% and 8.7% for menaquinone-4; and were 13.2%,11.1% and 7.2% for menaquinone-7. No interference was found between K1, menaquinone-4 and menaquinone-7, nor any deuterated internal standards. This method was then used to determine reference values for Caucasian populations of central European origin. Samples were measured from 191 healthy volunteers (51.2 ± 16.2 years (mean ± SD)) and the values concerning K1 were 0.044-1.357 ng/mL for women and 0.030-1.214 ng/mL for men. The values for menaquinone-4 and menaquinone-7 did not exhibit any differences between women and men, and were 0.050-1.598 and 0.074-0.759 ng/mL, respectively.

Department of Medical Chemistry and Clinical Biochemistry Charles University Prague 2nd Faculty of Medicine and University Hospital Motol Prague Czech Republic

Zobrazit více v PubMed

Aytekin M, Emerk K. Accurate reference intervals are required for accurate diagnosis and monitoring of patients. EJIFCC. 2008;19(2):137–141. PubMed PMC

Booth SL, Broe KE, Peterson JW, Cheng DM, Dawson-Hughes B, Gundberg CM, Cupples LA, Wilson PW, Kiel DP. Associations between vitamin K biochemical measures and bone mineral density in men and women. Journal of Clinical Endocrinology & Metabolism. 2004;89(10):4904–4909. doi: 10.1210/jc.2003-031673. PubMed DOI

EFSA NDA Panel (EFSA Panel on Dietetic Products, Nutrition and Allergies) Turck D, Bresson J-L, Burlingame B, Dean T, Fairweather-Tait S, Heinonen M, Hirsch-Ernst KI, Mangelsdorf I, McArdle HJ, Naska A, Nowicka G, Pentieva K, Sanz Y, Siani A, Sjödin A, Stern M, Tomé D, Van Loveren H, Vinceti M, Willatts P, Lamberg-Allardt C, Przyrembel H, Tetens I, Dumas C, Fabiani L, Ioannidou S, Neuhäuser-Berthold M. Scientific opinion on the dietary reference values for vitamin K. EFSA Journal. 2017;15(5):4780. PubMed PMC

Food and Agriculture Organization of the United Nations/World Health Organization Human vitamin and mineral requirements. 2001. http://www.fao.org/3/a-y2809e.pdf http://www.fao.org/3/a-y2809e.pdf Report of a joint FAO/WHO expert consultation. Bangkok.

Fu X, Peterson JW, Hdeib M, Booth SL, Grusak MA, Lichtenstein AH, Dolnikowski GG. Measurement of deuterium-labeled phylloquinone in plasma by high-performance liquid chromatography/mass spectrometry. Analytical Chemistry. 2009;81(13):5421–5425. doi: 10.1021/ac900732w. PubMed DOI PMC

Fusaro M, Crepaldi G, Maggi S, D’Angelo A, Calo L, Miozzo D, Fornasieri A, Gallieni M. Bleeding, vertebral fractures and vascular calcifications in patients treated with warfarin: hope for lower risks with alternative therapies. Current Vascular Pharmacology. 2011;9(6):763–769. doi: 10.2174/157016111797484134. PubMed DOI

Fusaro M, Gallieni M, Rizzo MA, Stucchi A, Delanaye P, Cavalier E, Moysés RMA, Jorgetti V, Iervasi G, Giannini S, Fabris F, Aghi A, Sella S, Galli F, Viola V, Plebani M. Vitamin K plasma levels determination in human health. Clinical Chemistry and Laboratory Medicine. 2017;55(6):789–799. doi: 10.1515/cclm-2016-0783. PubMed DOI

Harrell FE, Davis CE. A new distribution-free quantile estimator. Biometrika. 1982;69(3):635–640. doi: 10.1093/biomet/69.3.635. DOI

Harrell F, Jr, Dupont C. Hmisc: Harrell Miscellaneous 2017. 2017. https://cran.r-project.org/web/packages/Hmisc/Hmisc.pdf. [4 September 2019]. https://cran.r-project.org/web/packages/Hmisc/Hmisc.pdf

Horowitz GL. Defining, establishing, and verifying reference intervals in the clinical laboratory: approved guideline. Third Edition. Wayne: Clinical and Laboratory Standards Institute; 2008.

Jiang Y, Yang J, Yang C, Meng F, Zhou Y, Yu B, Khan M, Yang H. Vitamin K4 induces tumor cytotoxicity in human prostate carcinoma PC-3 cells via the mitochondria-related apoptotic pathway. Pharmazie. 2013;68(6):442–448. PubMed

Kamao M, Suhara Y, Tsugawa N, Okano T. Determination of plasma vitamin K by high-performance liquid chromatography with fluorescence detection using vitamin K analogs as internal standards. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 2005;816(1–2):41–48. doi: 10.1016/j.jchromb.2004.11.003. PubMed DOI

Karl JP, Fu X, Dolnikowski GG, Saltzman E, Booth SL. Quantification of phylloquinone and menaquinones in feces, serum, and food by high-performance liquid chromatography–mass spectrometry. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 2014;963:128–133. doi: 10.1016/j.jchromb.2014.05.056. PubMed DOI

Klapkova E, Cepova J, Dunovska K, Prusa R. Determination of vitamin K1, MK-4 and MK-7 in human serum of postmenopausal women by HPLC with fluorescence detection. Journal of Clinical Laboratory Analysis. 2018;32(5):e22381. doi: 10.1002/jcla.22381. PubMed DOI PMC

Lee S, Morris YJ, Gross LK, Porter EE, Ortiz-Colon F, Farrow JM, Waqar AS, Phifer EC, Kerdahi KF. Development and single-lab validation of an UHPLC-APCI-MS/MS method for vitamin K1 in infant formulas and other nutritional formulas. Journal of Regulatory Science. 2015;2:27–35.

Nakaoka E, Tanaka S, Onda K, Sugiyama K, Hirano T. Effects of Vitamin K3 and K5 on Daunorubicin-resistant human T lymphoblastoid leukemia cells. Anticancer Research. 2015;35(11):6041–6048. PubMed

Osman NA, El-Abd N, Nasrallah M. VKORC1 gene (vitamin K epoxide reductase) polymorphisms are associated with cardiovascular disease in chronic kidney disease patients on hemodialysis. Saudi Journal of Kidney Disease and Transplantation. 2016;27:908–915. PubMed

R Core Team . R: a Language and environment for statistical computing. Vienna: The R Foundation for Statistical Computing; 2015.

Riphagen IJ, Van Der Molen JC, Van Faassen M, Navis G, De Borst MH, Muskiet FA, De Jong WH, Bakker SJ, Kema IP. Measurement of plasma vitamin K1 (phylloquinone) and K2 (menaquinones-4 and -7) using HPLC-tandem mass spectrometry. Clinical Chemistry and Laboratory Medicine. 2016;54(7):1201–1210. doi: 10.1515/cclm-2015-0864. PubMed DOI

Sadowski JA, Hood SJ, Dallal GE, Garry PJ. Phylloquinone in plasma from elderly and young adults: factors influencing its concentration. American Journal of Clinical Nutrition. 1989;50(1):100–108. doi: 10.1093/ajcn/50.1.100. PubMed DOI

Schurgers LJ, Teunissen KJF, Knapen MHJ, Kwaijtall M, Van Diest R, Appels A, Reutelingsperger CP, Cleutjens JPM, Vermeer C. Novel conformation-specific antibodies against matrix gamma-carboxyglutamic acid (Gla) protein: undercarboxylated matrix Gla protein as marker for vascular calcification. Arteriosclerosis, Thrombosis, and Vascular Biology. 2005;25:1629–1633. PubMed

Schwalfenberg GK. Vitamins K1 and K2: the emerging group of vitamins required for human health. Journal of Nutrition and Metabolism. 2017;2017(18):1–6. doi: 10.1155/2017/6254836. PubMed DOI PMC

Song Q, Wen A, Ding L, Dai L, Yang L, Qi X. HPLC–APCI–MS for determination of vitamin K1 in human plasma: method and clinical application. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 2008;875(2):541–545. doi: 10.1016/j.jchromb.2008.10.009. PubMed DOI

Stafford DW. The vitamin K cycle. Journal of Thrombosis and Haemostasis. 2005;3(8):1873–1878. doi: 10.1111/j.1538-7836.2005.01419.x. PubMed DOI

Suhara Y, Kamao M, Tsugawa N, Okano T. Method for the determination of vitamin K homologues in human plasma using high-performance liquid chromatography-tandem mass spectrometry. Analytical Chemistry. 2005;77(3):757–763. doi: 10.1021/ac0489667. PubMed DOI

Suttie JW. Vitamin K and human nutrition. Journal of the American Dietetic Association. 1992;92(5):585–590. PubMed

Tate JR, Yen T, Jones GRD. Transference and validation of reference intervals. Clinical Chemistry. 2015;61(8):1012–1015. doi: 10.1373/clinchem.2015.243055. PubMed DOI

Tie J-K, Jin D-Y, Straight DL, Stafford DW. Functional study of the vitamin K cycle in mammalian cells. Blood. 2011;117(10):2967–2974. doi: 10.1182/blood-2010-08-304303. PubMed DOI PMC

Tie J-K, Jin D-Y, Tie K, Stafford DW. Evaluation of warfarin resistance using transcription activator-like effector nucleases-mediated vitamin K epoxide reductase knockout HEK293 cells. Journal of Thrombosis and Haemostasis. 2013;11(8):1556–1564. doi: 10.1111/jth.12306. PubMed DOI PMC

Tie J-K, Stafford DW. Structural and functional insights into enzymes of the vitamin K cycle. Journal of Thrombosis and Haemostasis. 2015;14(2):236–247. doi: 10.1111/jth.13217. PubMed DOI PMC

Torri S, Ikari Y, Tanabe K, Kakuta T, Hatori M, Shioi M, Shioi A, Okano T. Plasma phylloquinone, menaquinone-4 and menaquinone-7 levels and coronary artery calcification. Journal of Nutritional Science. 2016;5:e48. doi: 10.1017/jns.2016.20. PubMed DOI PMC

Tsugawa N, Shiraki M, Suhara Y, Kamao M, Tanaka K, Okano T. Vitamin K status of healthy Japanese women: age-related vitamin K requirement for γ-carboxylation of osteocalcin. American Journal of Clinical Nutrition. 2006;83(2):380–386. doi: 10.1093/ajcn/83.2.380. PubMed DOI

U.S. Food and Drug Administration (FDA) U.S. Department of Health and Human Services. 2013. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM368107.pdf http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM368107.pdf Draft guidance for industry: bioanalytical method validation (Revised)

Yan L, Zhou B, Greenberg D, Wang L, Nigdikar S, Prynne C, Prentice A. Vitamin K status of older individuals in northern China is superior to that of older individuals in the UK. British Journal of Nutrition. 2004;92(6):939–945. doi: 10.1079/BJN20041261. PubMed DOI

Vitamin K Status Based on K1, MK-4, MK-7, and Undercarboxylated Prothrombin Levels in Adolescent and Adult Patients with Cystic Fibrosis: A Cross-Sectional Study

Vitamin K - sources, physiological role, kinetics, deficiency, detection, therapeutic use, and toxicity