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

Kamao, Maya; Suhara, Yoshitomo; Tsugawa, Naoko; Okano, Toshio

doi:10.1016/j.jchromb.2004.11.003

Cited by 71 publications

(57 citation statements)

References 33 publications

(31 reference statements)

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“…Maximal g-glutamyl carboxylation was observed at a vitamin K1 concentration of ;10 nM, which is similar to recently reported plasma vitamin K1 concentrations in humans by Shea et al of 2.0 nM to 3.8 nM 20 or Kamao et al of 2.8 nM. 21 The observation that g-glutamyl carboxylation proceeded in the absence of exogenous vitamin K suggests the presence of an endogenous vitamin K vitamer in a component of the cell medium. We were, however, unable to detect vitamin K1 in the fetal bovine serum.…”

Section: Vitamin K Dependence and Warfarin Inhibition Of F9ch G-carbosupporting

confidence: 87%

A cellular system for quantitation of vitamin K cycle activity: structure-activity effects on vitamin K antagonism by warfarin metabolites

Haque

McDonald

Kulman

et al. 2014

Blood

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• Factor IX glutamyl carboxylation in engineered HEK 293 cells recapitulates in vivo anticoagulant inhibition of vitamin K cycle activity.• Warfarin metabolite structureactivity analysis on vitamin K cycle antagonism determines their contributions to in vivo anticoagulation.Warfarin and other 4-hydroxycoumarins inhibit vitamin K epoxide reductase (VKOR) by depleting reduced vitamin K that is required for posttranslational modification of vitamin K-dependent clotting factors. In vitro prediction of the in vivo potency of vitamin K antagonists is complicated by the complex multicomponent nature of the vitamin K cycle. Here we describe a sensitive assay that enables quantitative analysis of g-glutamyl carboxylation and its antagonism in live cells. We engineered a human embryonic kidney (HEK) 293-derived cell line (HEK 293-C3) to express a chimeric protein (F9CH) comprising the Gla domain of factor IX fused to the transmembrane and cytoplasmic regions of proline-rich Gla protein 2. Maximal g-glutamyl carboxylation of F9CH required vitamin K supplementation, and was dose-dependently inhibited by racemic warfarin at a physiologically relevant concentration. Cellular g-glutamyl carboxylation also exhibited differential VKOR inhibition by warfarin enantiomers (S > R ) consistent with their in vivo potencies. We further analyzed the structure-activity relationship for inhibition of g-glutamyl carboxylation by warfarin metabolites, observing tolerance to phenolic substitution at the C-5 and especially C-6, but not C-7 or C-8, positions on the 4-hydroxycoumarin nucleus. After correction for in vivo concentration and protein binding, 10-hydroxywarfarin and warfarin alcohols were predicted to be the most potent inhibitory metabolites in vivo. (Blood. 2014;123(4):582-589)

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Section: Vitamin K Dependence and Warfarin Inhibition Of F9ch G-carbosupporting

confidence: 87%

A cellular system for quantitation of vitamin K cycle activity: structure-activity effects on vitamin K antagonism by warfarin metabolites

Haque

McDonald

Kulman

et al. 2014

Blood

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“…The proposed HPLC-CL method was 2-1000 times more sensitive compared with the reported methods [21][22][23][24][25][26][27][28] and with similar sensitivity to LC-MS method [29]. However, the simplicity and low cost make the developed method more superior to LC-MS method in routine assay of vitamin K homologues.…”

Section: Resultsmentioning

confidence: 94%

“…To date, several publications were reported for the separation and determination of vitamins K in biological samples including high performance liquid chromatography (HPLC) with ultraviolet (UV) [19][20], fluorescence [21][22][23][24][25], electrochemical [26][27], chemiluminescence (CL) [28] and mass spectrometric detection [29]. In initial study to detect vitamin K homologues in plasma of RA patients, it was found that vitamin K concentrations were significantly lower in RA patients that could not be detected by many of the reported methods [19][20][21][22][23][24][25][26][27][28] while others remain expensive techniques for routine assay of vitamin K concentrations [29].…”

Section: Introductionmentioning

confidence: 99%

“…In initial study to detect vitamin K homologues in plasma of RA patients, it was found that vitamin K concentrations were significantly lower in RA patients that could not be detected by many of the reported methods [19][20][21][22][23][24][25][26][27][28] while others remain expensive techniques for routine assay of vitamin K concentrations [29]. Therefore, it is necessary to develop a highly sensitive and selective determination method designated for measurements of lower concentrations vitamin K homologues in the presence of any existed interferences from medications or plasma components.…”

Section: Introductionmentioning

confidence: 99%

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Selective chemiluminescence method for monitoring of vitamin K homologues in rheumatoid arthritis patients

Ahmed

Kishikawa

Ohyama

et al. 2011

Talanta

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Vitamin K is a fat-soluble vitamin involved in blood coagulation and bone metabolism. The detection and monitoring of vitamin K homologues in rheumatoid arthritis (RA) patients is a challenging problem due to the smaller concentrations of vitamin K and the presence of several interfering medications. Therefore, this study aimed to develop a new highly sensitive and selective chemiluminescence (CL) method designated to quantify vitamin K homologues in plasma of RA patients selectively including phylloquinone (PK, vitamin K 1 ), menaquinone-4 (MK-4, vitamin K 2 ) and menaquinone-7 (MK-7, vitamin K 2 ). The method was based on the unique photochemical properties of vitamin K homologues that were exploited for selective luminol CL reaction. The correlation coefficients of 0.998 or more were obtained in the concentration ranges of 0.1-100 ng mL -1 vitamin K homologues. The detection limits were 0.03-0.1 ng mL -1 in human plasma for vitamin K homologues. The developed HPLC-CL system was successfully applied for selective determination of vitamin K homologues in plasma of RA patients. The developed method may provide a useful tool for monitoring vitamin K homologues in different clinical studies such as RA, osteoporosis and hepatocellular carcinoma in which vitamin K is intervented.

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“…Initially, HPLC with ultraviolet detection (UV) was used to Post-column reduction is used to convert the quinone structure of vitamin K in the corresponding hydroquinones, measured in oxidation mode Liquid chromatography tandem mass spectrometry (LC-APCI-MS/MS) Provides higher sensitivity and selectivity in comparison with other techniques determinate PK plasma concentration. Afterwards, HPLC with fluorescence after post column reduction demonstrated greater sensitivity and selectivity than UV method [44,45], but it required extensive sample pre-purification to reduce the chromatographic interference induced by lipids [41].…”

Section: Determination Of Vitamin K Plasma Levels: Technical Aspectsmentioning

confidence: 99%

Vitamin K plasma levels determination in human health

Fusaro

Gallieni

Rizzo

et al. 2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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Vitamin K (phylloquinone or vitamin K 1 and menaquinones or vitamin K 2 ) plays an important role as a cofactor in the synthesis of hepatic blood coagulation proteins, but recently has also aroused an increasing interest for its action in extra-hepatic tissues, in particular in the regulation of bone and vascular metabolism. The accurate measurement of vitamin K status in humans is still a critical issue. Along with indirect assays, such as the undercarboxylated fractions of vitamin K-dependent proteins [prothrombin, osteocalcin (OC), and matrix gla protein], the direct analysis of blood levels of phylloquinone and menaquinones forms might be considered a more informative and direct method for assessing vitamin K status. Different methods for direct quantification of vitamin K serum levels are available. High-performance liquid chromatography (HPLC) methods coupled with post-column reduction procedures and fluorimetric or electrochemical detection are commonly used for food and blood analysis of phylloquinone, but they show some limitations when applied to the analysis of serum menaquinones because of interferences from triglycerides. Recent advancements include liquid chromatography tandem mass spectrometry (LCMS/MS) detection, which assures higher specificity. The optimization and standardization of these methods requires specialized laboratories. The variability of results observed in the available studies suggests the need for further investigations to obtain more accurate analytical results.

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

Cited by 71 publications

References 33 publications

A cellular system for quantitation of vitamin K cycle activity: structure-activity effects on vitamin K antagonism by warfarin metabolites

A cellular system for quantitation of vitamin K cycle activity: structure-activity effects on vitamin K antagonism by warfarin metabolites

Selective chemiluminescence method for monitoring of vitamin K homologues in rheumatoid arthritis patients

Vitamin K plasma levels determination in human health

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