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Quantitation of vitamin K in human milk

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Lipids

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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Abbreviations

HDN:

hemorrhagic disease of the newborn

HPLC:

high performance liquid chromatography

k′:

(Ve−Vo)/Vo(Ve=elution volume

Vo :

void volume

OD:

optical density

TBAP:

tetrabutyl ammonium perchlorate

UV:

ultraviolet

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Authors and Affiliations

  1. Department of Biochemistry and Family and Community Medicine, University of Arizona, 85721, Tucson, Arizona

    Louise M. Canfield, Jeanne Burr, Larry Clark & Daniel L. McGee

  2. Department of Pediatrics, Baylor College of Medicine, 77030, Houston, Texas

    Judy M. Hopkinson

  3. Process Development Department, Pitman-Moore, P.O. Box 207, 47808, Terre Haute, IN

    Anne F. Lima

  4. Department of Biochemistry, Texas A&M University, 77843, College Station, TX

    Gail S. Martin

  5. 11731 Wickhollow Lane, 77043, Houston, TX

    Kyoto Sugimoto

Authors
  1. Louise M. Canfield
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  2. Judy M. Hopkinson
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  3. Anne F. Lima
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  4. Gail S. Martin
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  5. Kyoto Sugimoto
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  6. Jeanne Burr
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  7. Larry Clark
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  8. Daniel L. McGee
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Canfield, L.M., Hopkinson, J.M., Lima, A.F. et al. Quantitation of vitamin K in human milk. Lipids 25, 406–411 (1990). https://doi.org/10.1007/BF02537985

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  • DOI: https://doi.org/10.1007/BF02537985

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