Determination of plasma Vitamin K by high-performance liquid chromatography with fluorescence detection using Vitamin K analogs as internal standards

A HPLC fluorescence determination method for Vitamin K derivatives (Vitamin K 1, phylloquinone, PK and K 2, menaquinones, MK-4 and MK-7) using post-column reduction and internal standards was developed. Selectivity and reproducibility were increased by optimized chromatography conditions and satisfa...

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Veröffentlicht in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2005-02, Vol.816 (1), p.41-48
Hauptverfasser: Kamao, Maya, Suhara, Yoshitomo, Tsugawa, Naoko, Okano, Toshio
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container_title Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
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creator Kamao, Maya
Suhara, Yoshitomo
Tsugawa, Naoko
Okano, Toshio
description A HPLC fluorescence determination method for Vitamin K derivatives (Vitamin K 1, phylloquinone, PK and K 2, menaquinones, MK-4 and MK-7) using post-column reduction and internal standards was developed. Selectivity and reproducibility were increased by optimized chromatography conditions and satisfactory precision and accuracy were attained by using synthetic internal standards. After addition of internal standards to plasma samples, lipids were extracted with ethanol and hexane. Chromatography was performed by isocratic reverse phase separation on a C18 column. Vitamin K derivatives were detected at 430 nm with excitation at 320 nm for MK-4 and 240 nm for PK and MK-7. The detection limits for MK-4, PK and MK-7 were 4, 2 and 4 pg, respectively. The recoveries of MK-4, PK and MK-7 were greater than 92% and the inter- and intra-assay R.S.D. values were 5.7–9.2% for MK-4, 4.9–9.6% for PK and 6.3–19.3% for MK-7. The data showed good correlation between proposed method and LC-APCI/MS method for MK-4 ( R 2 = 0.988), PK ( R 2 = 0.979) and MK-7 ( R 2 = 0.986). The method allows the determination of Vitamin K for evaluating their clinical and nutritional status.
doi_str_mv 10.1016/j.jchromb.2004.11.003
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B, Analytical technologies in the biomedical and life sciences</title><addtitle>J Chromatogr B Analyt Technol Biomed Life Sci</addtitle><description>A HPLC fluorescence determination method for Vitamin K derivatives (Vitamin K 1, phylloquinone, PK and K 2, menaquinones, MK-4 and MK-7) using post-column reduction and internal standards was developed. Selectivity and reproducibility were increased by optimized chromatography conditions and satisfactory precision and accuracy were attained by using synthetic internal standards. After addition of internal standards to plasma samples, lipids were extracted with ethanol and hexane. Chromatography was performed by isocratic reverse phase separation on a C18 column. Vitamin K derivatives were detected at 430 nm with excitation at 320 nm for MK-4 and 240 nm for PK and MK-7. The detection limits for MK-4, PK and MK-7 were 4, 2 and 4 pg, respectively. 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subjects Analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, High Pressure Liquid - methods
Fluorescence
Fluorescence detection
Fundamental and applied biological sciences. Psychology
General pharmacology
Humans
Internal standards
Medical sciences
Osteoporosis - blood
Osteoporosis - drug therapy
Pharmacology. Drug treatments
Reference Standards
Spectrometry, Mass, Electrospray Ionization
Vitamin K
Vitamin K - analogs & derivatives
Vitamin K - blood
Vitamin K 2 - analogs & derivatives
Vitamin K 2 - blood
Vitamin K 2 - therapeutic use
title Determination of plasma Vitamin K by high-performance liquid chromatography with fluorescence detection using Vitamin K analogs as internal standards
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