Sensitive and simple determination of vitamin K1 in human plasma by ultra-performance liquid chromatography coupled with a fluorescence detector
Izumi Ohno, Takuya Araki, Masashi Kawano, Tomonori Nakamura, Koujirou Yamamoto
We developed a sensitive and simple method for the determination of vitamin K1 (VK1) in human plasma using ultra-performance liquid chromatography with post-column reduction. VK1 was extracted with single-step liquid-liquid partitioning. As an analytical column, we used an ACQUITY CSH C18 (1.7 μm, 2.1 × 50 mm). The mobile phase consisted of a mixture of distilled water/methanol delivered using a linear gradient, starting with a composition of 94% methanol and ending at 100% methanol with a flow rate of 0.6 mL/min. Commonly, synthetic vitamin K analogs are used as the internal standards (IS); however, we selected menaquinone-8, which is undetectable in human plasma. The peaks were detected by a fluorescence detector (excitation at 244 nm, emission at 430 nm). The lower limit of quantification was 0.1 ng/mL. The inter- and intra-assay precision varied between 2.7-16.1%, and the accuracy varied between -10.3-13.4%. This method was simple, rapid, and sensitive for the detection of VK1 and is suitable for the determination of VK1 in small volumes of human serum. We believe that this method is useful for determining the concentration of VK1 in human plasma and in pharmacodynamic studies of warfarin, a VK antagonist, with clinical samples.
Journal of Liquid Chromatography and Related Technologies
Fluorescence detector
Post-column reduction
UPLC
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Vitamin K 1 Medicine & Life Sciences 100%
Ultra Performance Liquid Chromatography Chemical Compounds 84%
Liquid Chromatography Medicine & Life Sciences 71%
Vitamin Chemical Compounds 69%
Fluorescence Medicine & Life Sciences 55%
Methanol Medicine & Life Sciences 42%
Menaquinone-8 Chemical Compounds 28%
Ohno, I., Araki, T., Kawano, M., Nakamura, T., & Yamamoto, K. (2013). Sensitive and simple determination of vitamin K1 in human plasma by ultra-performance liquid chromatography coupled with a fluorescence detector. Journal of Liquid Chromatography and Related Technologies, 36(8), 1068-1077. https://doi.org/10.1080/10826076.2012.685908
Sensitive and simple determination of vitamin K1 in human plasma by ultra-performance liquid chromatography coupled with a fluorescence detector. / Ohno, Izumi; Araki, Takuya; Kawano, Masashi et al.
In: Journal of Liquid Chromatography and Related Technologies, Vol. 36, No. 8, 01.03.2013, p. 1068-1077.
Ohno, I, Araki, T, Kawano, M, Nakamura, T & Yamamoto, K 2013, 'Sensitive and simple determination of vitamin K1 in human plasma by ultra-performance liquid chromatography coupled with a fluorescence detector', Journal of Liquid Chromatography and Related Technologies, vol. 36, no. 8, pp. 1068-1077. https://doi.org/10.1080/10826076.2012.685908
Ohno I, Araki T, Kawano M, Nakamura T, Yamamoto K. Sensitive and simple determination of vitamin K1 in human plasma by ultra-performance liquid chromatography coupled with a fluorescence detector. Journal of Liquid Chromatography and Related Technologies. 2013 Mar 1;36(8):1068-1077. doi: 10.1080/10826076.2012.685908
Ohno, Izumi ; Araki, Takuya ; Kawano, Masashi et al. / Sensitive and simple determination of vitamin K1 in human plasma by ultra-performance liquid chromatography coupled with a fluorescence detector. In: Journal of Liquid Chromatography and Related Technologies. 2013 ; Vol. 36, No. 8. pp. 1068-1077.
@article{a6e79832e6e94586b49d3c209543d11a,
title = "Sensitive and simple determination of vitamin K1 in human plasma by ultra-performance liquid chromatography coupled with a fluorescence detector",
abstract = "We developed a sensitive and simple method for the determination of vitamin K1 (VK1) in human plasma using ultra-performance liquid chromatography with post-column reduction. VK1 was extracted with single-step liquid-liquid partitioning. As an analytical column, we used an ACQUITY CSH C18 (1.7 μm, 2.1 × 50 mm). The mobile phase consisted of a mixture of distilled water/methanol delivered using a linear gradient, starting with a composition of 94% methanol and ending at 100% methanol with a flow rate of 0.6 mL/min. Commonly, synthetic vitamin K analogs are used as the internal standards (IS); however, we selected menaquinone-8, which is undetectable in human plasma. The peaks were detected by a fluorescence detector (excitation at 244 nm, emission at 430 nm). The lower limit of quantification was 0.1 ng/mL. The inter- and intra-assay precision varied between 2.7-16.1%, and the accuracy varied between -10.3-13.4%. This method was simple, rapid, and sensitive for the detection of VK1 and is suitable for the determination of VK1 in small volumes of human serum. We believe that this method is useful for determining the concentration of VK1 in human plasma and in pharmacodynamic studies of warfarin, a VK antagonist, with clinical samples.",
keywords = "Fluorescence detector, Liquid-liquid extraction, MK-8, Post-column reduction, UPLC, Vitamin K1",
author = "Izumi Ohno and Takuya Araki and Masashi Kawano and Tomonori Nakamura and Koujirou Yamamoto",
note = "Funding Information: This study was supported by a grant-in-aid for scientific research KAKENHI 21590150 from the Japan Society for the Promotion of Science. Koujirou Yamamoto has received a donation from Eisai Co., Ltd. (Tokyo, Japan). All other authors have no conflict of interest to declare. We thank Ms. Aiko Matsumoto (Gunma University) for her secretarial assistance.",
journal = "Journal of Liquid Chromatography and Related Technologies",
T1 - Sensitive and simple determination of vitamin K1 in human plasma by ultra-performance liquid chromatography coupled with a fluorescence detector
AU - Ohno, Izumi
AU - Araki, Takuya
AU - Kawano, Masashi
AU - Nakamura, Tomonori
AU - Yamamoto, Koujirou
N1 - Funding Information: This study was supported by a grant-in-aid for scientific research KAKENHI 21590150 from the Japan Society for the Promotion of Science. Koujirou Yamamoto has received a donation from Eisai Co., Ltd. (Tokyo, Japan). All other authors have no conflict of interest to declare. We thank Ms. Aiko Matsumoto (Gunma University) for her secretarial assistance.
N2 - We developed a sensitive and simple method for the determination of vitamin K1 (VK1) in human plasma using ultra-performance liquid chromatography with post-column reduction. VK1 was extracted with single-step liquid-liquid partitioning. As an analytical column, we used an ACQUITY CSH C18 (1.7 μm, 2.1 × 50 mm). The mobile phase consisted of a mixture of distilled water/methanol delivered using a linear gradient, starting with a composition of 94% methanol and ending at 100% methanol with a flow rate of 0.6 mL/min. Commonly, synthetic vitamin K analogs are used as the internal standards (IS); however, we selected menaquinone-8, which is undetectable in human plasma. The peaks were detected by a fluorescence detector (excitation at 244 nm, emission at 430 nm). The lower limit of quantification was 0.1 ng/mL. The inter- and intra-assay precision varied between 2.7-16.1%, and the accuracy varied between -10.3-13.4%. This method was simple, rapid, and sensitive for the detection of VK1 and is suitable for the determination of VK1 in small volumes of human serum. We believe that this method is useful for determining the concentration of VK1 in human plasma and in pharmacodynamic studies of warfarin, a VK antagonist, with clinical samples.
AB - We developed a sensitive and simple method for the determination of vitamin K1 (VK1) in human plasma using ultra-performance liquid chromatography with post-column reduction. VK1 was extracted with single-step liquid-liquid partitioning. As an analytical column, we used an ACQUITY CSH C18 (1.7 μm, 2.1 × 50 mm). The mobile phase consisted of a mixture of distilled water/methanol delivered using a linear gradient, starting with a composition of 94% methanol and ending at 100% methanol with a flow rate of 0.6 mL/min. Commonly, synthetic vitamin K analogs are used as the internal standards (IS); however, we selected menaquinone-8, which is undetectable in human plasma. The peaks were detected by a fluorescence detector (excitation at 244 nm, emission at 430 nm). The lower limit of quantification was 0.1 ng/mL. The inter- and intra-assay precision varied between 2.7-16.1%, and the accuracy varied between -10.3-13.4%. This method was simple, rapid, and sensitive for the detection of VK1 and is suitable for the determination of VK1 in small volumes of human serum. We believe that this method is useful for determining the concentration of VK1 in human plasma and in pharmacodynamic studies of warfarin, a VK antagonist, with clinical samples.
KW - Fluorescence detector
KW - Liquid-liquid extraction
KW - MK-8
KW - Post-column reduction
KW - UPLC
KW - Vitamin K1
JO - Journal of Liquid Chromatography and Related Technologies
JF - Journal of Liquid Chromatography and Related Technologies