Measurement of menadione in urine by HPLC

J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Sep 15;878(26):2457-60. doi: 10.1016/j.jchromb.2010.07.016. Epub 2010 Jul 29.

Abstract

Menadione is a metabolite of vitamin K that is excreted in urine. A high performance liquid chromatography (HPLC) method using a C(30) column, post-column zinc reduction and fluorescence detection was developed to measure urinary menadione. The mobile phase was composed of 95% methanol with 0.55% aqueous solution and 5% DI H(2)O. Menaquinone-2 (MK-2) was used as an internal standard. The standard calibration curve was linear with a correlation coefficient (R(2)) of 0.999 for both menadione and MK-2. The lower limit of quantification (LLOQ) was 0.3pmole menadione/mL urine. Sample preparation involved hydrolysis of menadiol conjugates and oxidizing the released menadiol to menadione. Using this method, urinary menadione was shown to increase in response to 3 years of phylloquinone supplementation. This HPLC method is a sensitive and reproducible way to detect menadione in urine.

Trial registration: ClinicalTrials.gov NCT00183001.

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Clinical Trials as Topic
  • Drug Stability
  • Humans
  • Linear Models
  • Methanol / chemistry
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Vitamin K 1 / administration & dosage
  • Vitamin K 2 / analysis
  • Vitamin K 3 / urine*
  • Water / chemistry

Substances

  • Water
  • Vitamin K 2
  • Vitamin K 3
  • Vitamin K 1
  • Methanol

Associated data

  • ClinicalTrials.gov/NCT00183001