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The Content of Phylloquinone (Vitamin K₁) in Human Milk, Cows' Milk and Infant Formula Foods Determined by High-Performance Liquid Chromatography¹

Department of Haematology, Clinical Science Laboratories, Guy's Tower (17th and 18th floors), Cuy's Hospital, London, SE1 9RT, U. K.

Phylloquinone (2-methyl-3-phytyl-1,4-naphthoquinone) in human and cow's milk and in infant formula foods has been assayed by a method based on highperformance liquid chromatography (HPLC). The method has three chromatographic steps consisting of a preliminary purification of lipid extracts by conventional liquid chromatography, a further fractionation by semipreparative HPLC and a final analytical step by reversed-phase HPLC in which phylloquinone was resolved from the remaining contaminants and quantified by reference to an internal standard (phylloquinone 2,3-epoxide). The identity of the chromatographic peak ascribed to phylloquinone (vitamin K₁) was established by mass spectrometry. Mature human milk from 20 lactating mothers gave a mean concentration of phylloquinone of 2.1 µg/liter, and colostrum from 9 mothers gave a mean value of 2.3 µg/liter. These levels in human milk were significantly lower than those found in either Friesian (Holstein) cows' milk (mean 4.9 µg/liter) or unsupplemented infant formula foods containing only cows' milk fat (mean 4.2 µg/liter). The mean phylloquinone content of two unsupplemented infant formula foods containing only vegetable oils was 11.5 µg/liter. After an oral dose of 20 mg phylloquinone, the concentration of K₁ in the breast milk of one mother rose to 140 µg/liter after 12 hours and at 48 hours was still about twice the average endogenous level of human milk.

KEY WORDS: • vitamin K • high-performance liquid chromatography • milk • infant formulas

¹ These studies were supported by a grant (G977/827) from the Medical Research Council, U. K. A preliminary report of the studies contained in this paper was presented at a meeting of the Nutrition Society; see Haroon, Y., Shearer, M. J., McEnery, G., Alian, V. E. & Barkhan, P. (1980) Proc. Nutr. Soc. 39, 49A.

² Present address: Harvard Medical School, Department of Orthopedic Surgery, The Children's Hospital Medical Center, 300 Longwood Avenue, Boston. MA 02115.

³ Department of Chemistry, Queen Elizabeth College, University of London, Campden Hill Road, London, U. K.

⁴ Pediatric Department, Whipps Cross Hopsital, London, U. K.

Manuscript received 30 October 1981.

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