Measurement of menadione in urine by HPLC.
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.
Al Rajabi
Ala
A
USDA Human Nutrition Research Center on Aging at Tufts University, 711 Washington Street, Boston, MA 02111, USA. ala.al rajabi@tufts.edu
Peterson
James
J
Choi
Sang-Woon
SW
Suttie
John
J
Barakat
Susan
S
Booth
Sarah L
SL
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002986:Clinical Trials as Topic; D004355:Drug Stability; D006801:Humans; D016014:Linear Models; D000432:Methanol / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D010837:Vitamin K 1 / Q000008:administration & dosage; D024482:Vitamin K 2 / Q000032:analysis; D024483:Vitamin K 3 / Q000652:urine*; D014867:Water / Q000737:chemistry
D014867:Water; D024482:Vitamin K 2; D024483:Vitamin K 3; D010837:Vitamin K 1; D000432:Methanol