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一种用于测定发酵液中维生素K2-7的新型快速高效液相色谱-紫外检测法。

A Novel Fast HPLC-UV Method for Menaquinone-7 Determination in Fermentation Broth.

作者信息

Celiento Alessia, Giuliani Antonella, Laini Michela, Longoni Marco, Rizzi Giuseppina, Dei Nobili Mariemma

机构信息

R&D Analytical Development Department, Gnosis by Lesaffre S.p.A, Desio 20832, Italy.

出版信息

ACS Omega. 2025 Apr 28;10(18):18793-18801. doi: 10.1021/acsomega.5c00483. eCollection 2025 May 13.

Abstract

Menaquinone-7 (MK-7) plays a key role in bone and cardiovascular health, and it is often included in multivitamin supplements and functional foods. The aim of this study is the development and validation, according to ICH guidelines, of a fast, highly sensitive, and reliable reverse-phase HPLC-UV quantification of vitamin K as MK-7 in a common biological matrix sample, such as fermentation broth medium from , with the advantage of single-step extraction and short analysis time. MK-7 was successfully isolated from the fermentation broth and its interfering residues by thermo-acidic extraction using 5% HSO and ethanol, without derivatization, and was retained using a C8 column. The isocratic elution and UV detection at 268 nm within a 3-min run time allowed quantification over a wide accuracy range of 0.10-18.00 μg/mL, with an LOQ value of 0.10 μg/mL and an LOD value of 0.03 μg/mL. The method was found to be precise, accurate, and reliable, with RSD% lower than 5% and recoveries varying between 96.0% and 108.9%. The proposed method unlocks the opportunity to be suitable for routine analysis in the screening of bacterial strains producing MK-7, as well as to be successfully employed in manufacturing processes in the nutraceutical and food industries where complex matrices are involved.

摘要

甲萘醌-7(MK-7)在骨骼和心血管健康方面起着关键作用,并且它经常被包含在多种维生素补充剂和功能性食品中。本研究的目的是根据国际人用药品注册技术协调会(ICH)指南,开发并验证一种快速、高度灵敏且可靠的反相高效液相色谱-紫外(HPLC-UV)方法,用于在常见生物基质样品(如来自[具体来源未给出]的发酵肉汤培养基)中对作为MK-7的维生素K进行定量分析,该方法具有一步提取和分析时间短的优点。通过使用5%硫酸和乙醇进行热酸性萃取,成功地从发酵肉汤及其干扰性残留物中分离出MK-7,无需衍生化,并使用C8柱进行保留。在3分钟的运行时间内,采用等度洗脱并在268nm处进行紫外检测,可在0.10 - 18.00μg/mL的宽准确度范围内进行定量分析,定量限(LOQ)值为0.10μg/mL,检测限(LOD)值为0.03μg/mL。该方法被发现具有精密度高、准确度高和可靠性好的特点,相对标准偏差(RSD%)低于5%,回收率在96.0%至108.9%之间变化。所提出的方法为筛选产生MK-7的细菌菌株进行常规分析提供了机会,并且能够成功应用于涉及复杂基质的营养保健品和食品工业的生产过程中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d288/12079229/c3ec5b16355e/ao5c00483_0001.jpg

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