Measurement of menadione in urine by HPLC |
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| Authors: | Ala Al Rajabi James Peterson Sang-Woon Choi John Suttie Susan Barakat Sarah L Booth |
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| Institution: | 1. USDA Human Nutrition Research Center on Aging at Tufts University, 711 Washington Street, Boston, MA 02111, USA;2. Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53706, USA |
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| Abstract: | Menadione is a metabolite of vitamin K that is excreted in urine. A high performance liquid chromatography (HPLC) method using a C30 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 H2O. Menaquinone-2 (MK-2) was used as an internal standard. The standard calibration curve was linear with a correlation coefficient (R2) of 0.999 for both menadione and MK-2. The lower limit of quantification (LLOQ) was 0.3 pmole 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. |
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| Keywords: | MD menadione MK-4 menaquinone-4 MK-2 menaquinone-2 GGPP geranylgeranyl pyrophosphate PK phylloquinone 7-EC 7-ethoxycoumarin LLOQ lower limit of quantification |
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