Development of a liquid chromatography mass spectrometry method for the determination of vitamin K1, menaquinone-4, menaquinone-7 and vitamin K1-2,3 epoxide in serum of individuals without vitamin K supplements
Author:
Meinitzer Andreas1, Enko Dietmar12, Zelzer Sieglinde1, Prüller Florian1, Alonso Nerea1, Fritz-Petrin Eva1, Herrmann Markus1
Affiliation:
1. Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz , Graz , Austria 2. Institute of Clinical Chemistry and Laboratory Medicine, General Hospital Hochsteiermark , Leoben , Austria
Abstract
Abstract
Objectives
Vitamin K and metabolites have a beneficial role in blood coagulation, bone metabolism and growth. However, the determination of vitamin K concentrations in the blood in patients consuming a diet with naturally occurring vitamin K is currently challenging. We aim to develop a cost-effective and rapid method to measure vitamin K metabolites with potential application for clinics and research.
Methods
We developed a simple liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of vitamin K1, menaquinone-4 (MK-4), menaquinone-7 (MK-7) and vitamin K1-2,3 epoxide in human serum and validated the method in a study cohort of 162 patients tested for carbohydrate malabsorption and in 20 patients with oral phenprocoumon intake.
Results
The overall precision (CVs) ranged between 4.8 and 17.7% in the specified working range (0.06–9.0 nmol/L for all analytes except for MK-7 with 0.04–6.16 nmol/L). In the malabsorption cohort samples, measured values were obtained for all different vitamin K metabolites except for vitamin K1-2,3 epoxide. This metabolite could be detected only in patients with phenprocoumon intake. The good performance of the method is especially achieved by the interaction of three factors: the use of lipase in the sample preparation, the use of an atypical fluorinated reversed phase column, and a logarithmic methanol gradient.
Conclusions
The described method is able to determine the concentration of four vitamin K metabolites in a time-efficient, simple and cost-effective manner. It can be suitable for both routine clinics and research.
Publisher
Walter de Gruyter GmbH
Subject
Biochemistry (medical),Clinical Biochemistry,General Medicine
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