A Validated HPLC Method for the Determination of Vitamin K in Human Serum – First Application in a Pharmacological Study

Marinova, Milka; Lütjohann, Dieter; Westhofen, P.; Watzka, Matthias; Breuer, Olof; Oldenburg, J.

doi:10.2174/1874241601104010017

Cited by 17 publications

(19 citation statements)

References 37 publications

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“…F p.o. values were in accordance with data from previous studies [28] and revealed substantial inter-individual variability. It is noteworthy that the highest bioavailability was found in heterozygotes, carriers of the AG genotype; furthermore, it was higher for male than for female subjects, although this finding cannot be physiologically and clinically eligible.…”

Section: Discussionsupporting

confidence: 92%

“…The limits of detection and quantification were 0.015 ng mL −1 and 0.15 ng mL −1 respectively. High sensitivity, analytical recoveries, accuracy, and calibration curve linearities could be reached, as shown before [28]. The within-day and day-to-day coefficients of variation amounted to less than 10 %, while the recovery ranged from 91 % to 114 %.…”

Section: Methodssupporting

confidence: 69%

“…Vitamin K1 was determined by reversed phase HPLC combined with fluorometric detection following post-column zinc reduction as described previously [28]. Briefly, the vitamin K related compounds were purified from serum by liquid–liquid extraction using n -hexane.…”

Section: Methodsmentioning

confidence: 99%

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VKORC1-dependent pharmacokinetics of intravenous and oral phylloquinone (vitamin K1) mixed micelles formulation

Marinova¹,

Lütjohann²,

Breuer³

et al. 2012

Eur J Clin Pharmacol

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ObjectiveThe pharmacokinetics of phylloquinone (vitamin K1) were evaluated in healthy human adult volunteers (15 male and 15 female) following oral and intravenous administration of a mixed micelles formulation (Konakion® MM 2 mg) in an open label study design. The subjects were allocated to one of three genotype-specific groups (n = 10 in each group) in terms of VKORC1 promoter polymorphism c.-1639 G > A to explore the relationship between genotype and pharmacokinetic parameters.MethodsBlood samples were collected for up to 24 h after administration. Phylloquinone serum levels were determined by reversed phase HPLC with fluorometric detection after post-column zinc reduction. Pharmacokinetic evaluation was performed using non-compartmental analysis.ResultsPharmacokinetic analysis of serum phylloquinone concentration versus time profiles revealed significant differences in the main pharmacokinetic parameters between groups. Upon oral administration, VKORC1 AG carriers showed 41 % higher mean bioavailability (p = 0.01) compared with homozygous AA individuals. Furthermore, AG subjects exhibited 30 % (p = 0.042) and 36 % (p = 0.021) higher mean AUC compared with GG and AA respectively. Terminal half-life was 32 % and 27 % longer for AG carriers in comparison to GG (p = 0.004) and AA (p = 0.015) genotypes respectively.ConclusionPharmacokinetic differences indicated significant inter-individual variance of vitamin K fate in the human body. The influence of the VKORC1 promoter polymorphism c.-1639 G > A on the pharmacokinetic properties of phylloquinone could be demonstrated in humans. To gain deeper insight in other potential genetic determinants of systemic vitamin K exposure, further correlation of the phenotype–genotype relationship of different players in vitamin K turnover has to be gained.

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Section: Discussionsupporting

confidence: 92%

Section: Methodssupporting

confidence: 69%

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VKORC1-dependent pharmacokinetics of intravenous and oral phylloquinone (vitamin K1) mixed micelles formulation

Marinova¹,

Lütjohann²,

Breuer³

et al. 2012

Eur J Clin Pharmacol

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“…4°C) in amber glass bottles or vials based upon a published literature report that demonstrates stability for at least 60 days when stored under these conditions. 6 Validation Sample Matrix. Canola seed used for method development and validation was purchased from a commercial source.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Measurement of Vitamin K₁ in Commercial Canola Cultivars from Growing Locations in North and South America Using High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Claussen

Taylor

Breeze

et al. 2015

J. Agric. Food Chem.

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Plant oils, including canola oil, are considered to be major sources of vitamin K as the second most substantial contributor of vitamin K to the human diet. Green leafy vegetables are the largest source of vitamin K. However, the effects of environment and germplasm on vitamin K levels in harvested canola seed have not been extensively investigated. To better understand these relationships, harvested canola seed from a range of diverse cultivars grown in different geographical locations in North and South America was assessed for levels of vitamin K. The analytical method developed to perform this measurement was based on C reversed-phase HPLC that could distinguish the biologically active trans-vitamin K from the inactive cis-isomer. Results demonstrated that for the majority of the canola cultivars evaluated, those cultivated in the North American sites had higher average vitamin K levels than those cultivated in the South American sites. Not all of the cultivars exhibited differences in response to the environment, suggesting that individual cultivar genetics also played a role in the variability of vitamin K levels observed in canola seed. Results from this study suggest that cultivar and environmental effects influence vitamin K levels in canola seed and provide a context to assess compositional variability of new cultivars.

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“…The present study reveals that nutraceutical formulation with vitamins was chosen to develop analytical separation method using Soxhlet's extraction with acetone solvent followed by evaporation with a rotary evaporator and fluorescence detection [14][15][16][17] . The developed method is precise, linear, accurate, robust and stability indicating compared to other UV detection regular HPLC methods.…”

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confidence: 99%

Vitamin K2-4 and K2-7 Estimation in Nutraceutical Solid Dosage Forms by Post Column Derivatization with Fluorescence Detection

Karuppiah¹,

Basha²

2016

ijps

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A post column derivatization method for the estimation of fat soluble vitamins K 2 -4 and K 2 -7 was developed by reverse phase HPLC and validated as per ICH guidelines. The compounds were extracted by solvent extraction with acetone followed by evaporation in a rotary evaporator. The residue was dissolved in ethanol and injected in to a chromatograph consisting of C18 column (Waters symmetry, 150×4.6, 3.5 μ) and fluorescence detector. The derivatization reagent was prepared by dissolving 136 mg of zinc chloride, 40 mg of sodium acetate and 0.1 ml of glacial acetic acid in methanol. The mobile phase comprises of methanol:isopropyl alcohol:acetonitrile:zinc chloride buffer solution 850:90:50:10 with a flow rate of 1 ml/min. The overall percentage recoveries of five different levels were found to be 99.85 and 100.5%, respectively. The linearity of the analytical method was determined from 10% to 120% level and the linear regression coefficients were 0.9991 and 0.9995 which is well within the acceptance criteria of 0.999. The limits of detection and limits of quantification were determined based on signal to noise ratio. The established values were 0.050, 0.50 μg/ml and 0.005, 0.047 μg/ml which is much lesser than available literatue limits. The developed method can be more suitable for the estimation of K 2 -4 and K 2 -7 present in drug substances as well as drug product formulations.Key words: Post column derivatization, nutraceutical solid dosage forms, estimation of K 2 -4 and K 2 -7 by HPLC, zinc chloride buffer solution, analytical estimation Vitamin K 2 includes several subtypes; the two subtypes most studied are menaquinone-4 (menatetrenone, MK-4) and menaquinone-7 (MK-7). Vitamin K 2 -4 ( fig. 1a) is yellow coloured powder, molecular formula C 31 H 40 O 2, molecular weight 444.65 g/mol, soluble in acetone and ethyl alcohol [1] . Vitamin K 2 -7 ( fig. 1b) is light yellow microcrystalline powder, molecular formula C 46 H 64 O 2, molecular weight 648.99 g/mol, soluble in petroleum ether, acetone, ethyl alcohol and insoluble in water [1] . Vitamin K is used for the treatment of anticoagulantinduced prothrombin deficiency caused by warfarin [2][3][4] . The American Academy of Paediatrics recommends that vitamin K should be given to all new born babies as a single intramuscular dose to prevent vitamin K deficiency bleeding. In Japan, vitamin K 2 is used for the management of osteoporosis. The fermented soya product nattō is rich in menaquinone-7 [2][3][4] .Derivatization reaction with the reduction of zinc metal ions to isolate compounds of closely eluting peaks by chromatography is fast emerging tool for analytical separation. The dosage form formulated with two active ingredients such as K 2 -4 and K 2 -7 along with excipients is very challenging effort to separate from its moiety. In order to separate the K series vitamins with almost same structural properties, the post column derivatization technique with fluorescence detection was used. The reduction mechanism of components with zinc chloride derivatizatio...

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A Validated HPLC Method for the Determination of Vitamin K in Human Serum – First Application in a Pharmacological Study

Cited by 17 publications

References 37 publications

VKORC1-dependent pharmacokinetics of intravenous and oral phylloquinone (vitamin K1) mixed micelles formulation

VKORC1-dependent pharmacokinetics of intravenous and oral phylloquinone (vitamin K1) mixed micelles formulation

Measurement of Vitamin K₁ in Commercial Canola Cultivars from Growing Locations in North and South America Using High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Vitamin K2-4 and K2-7 Estimation in Nutraceutical Solid Dosage Forms by Post Column Derivatization with Fluorescence Detection

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A Validated HPLC Method for the Determination of Vitamin K in Human Serum – First Application in a Pharmacological Study

Cited by 17 publications

References 37 publications

VKORC1-dependent pharmacokinetics of intravenous and oral phylloquinone (vitamin K1) mixed micelles formulation

VKORC1-dependent pharmacokinetics of intravenous and oral phylloquinone (vitamin K1) mixed micelles formulation

Measurement of Vitamin K1 in Commercial Canola Cultivars from Growing Locations in North and South America Using High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Vitamin K2-4 and K2-7 Estimation in Nutraceutical Solid Dosage Forms by Post Column Derivatization with Fluorescence Detection

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Product

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Measurement of Vitamin K₁ in Commercial Canola Cultivars from Growing Locations in North and South America Using High-Performance Liquid Chromatography–Tandem Mass Spectrometry