Quantitation of vitamin K in human milk |
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| Authors: | Louise M Canfield Judy M Hopkinson Anne F Lima Gail S Martin Kyoto Sugimoto Jeanne Burr Larry Clark Daniel L McGee |
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| Affiliation: | (1) Department of Biochemistry and Family and Community Medicine, University of Arizona, 85721 Tucson, Arizona;(2) Department of Pediatrics, Baylor College of Medicine, 77030 Houston, Texas;(3) Process Development Department, Pitman-Moore, P.O. Box 207, 47808 Terre Haute, IN;(4) Department of Biochemistry, Texas A&M University, 77843 College Station, TX;(5) 11731 Wickhollow Lane, 77043 Houston, TX |
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| Abstract: | A quantitative method was developed for the assay of vitamin K in human colostrum and milk. The procedure combines preparative
and analytical chromatography on silica gel in a nitrogen atmosphere followed by reversed phase high performance liquid chromatography
(HPLC). Two HPLC steps were used: gradient separation with ultraviolet (UV) detection followed by isocratic separation detected
electrochemically. Due to co-migrating impurities, UV detection alone is insufficient for identification of vitamin K. Exogenous
vitamin K was shown to equilibrate with endogenous vitamin K in the samples. A statistical method was incorporated to control
for experimental variability. Vitamin K1 was analyzed in 16 pooled milk samples from 7 donors and in individual samples from 15 donors at 1 month post-partrum. Vitamin
K1 was present at 2.94±1.94 and 3.15±2.87 ng/mL in pools and in individuals, respectively. Menaquinones, the bacterial form
of the vitamin, were not detected. The significance of experimental variation to studies of vitamin K in individuals is discussed. |
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