Lipids

, Volume 25, Issue 7, pp 406–411 | Cite as

Quantitation of vitamin K in human milk

  • Louise M. Canfield
  • Judy M. Hopkinson
  • Anne F. Lima
  • Gail S. Martin
  • Kyoto Sugimoto
  • Jeanne Burr
  • Larry Clark
  • Daniel L. McGee
Methods

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.

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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Copyright information

© American Oil Chemists’ Society 1990

Authors and Affiliations

  • Louise M. Canfield
    • 1
  • Judy M. Hopkinson
    • 2
  • Anne F. Lima
    • 3
  • Gail S. Martin
    • 4
  • Kyoto Sugimoto
    • 5
  • Jeanne Burr
    • 1
  • Larry Clark
    • 1
  • Daniel L. McGee
    • 1
  1. 1.Department of Biochemistry and Family and Community MedicineUniversity of ArizonaTucson
  2. 2.Department of PediatricsBaylor College of MedicineHouston
  3. 3.Process Development DepartmentPitman-MooreTerre Haute
  4. 4.Department of BiochemistryTexas A&M UniversityCollege Station
  5. 5.Houston

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