新型 7α-hydroxy-4-cholesten-3-one (C4) 液相色谱串联质谱法的开发与验证及其在评估分析前稳定性方面的实用性。

IF 3.8 2区 医学 Q1 MEDICAL LABORATORY TECHNOLOGY
Jonathan S Atkins, Brian G Keevil, Angela E Taylor, Christian Ludwig, James M Hawley
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引用次数: 0

摘要

目的:7α-羟基-4-胆甾烯-3-酮(C4)是两种初级胆汁酸的共同中间体。英国胃肠病学会建议将 C4 用于慢性腹泻患者胆汁酸腹泻(BAD)的检查。该项目旨在开发和验证一种测定方法,以定量检测血清中的 C4,并评估 C4 在未分离血液中的稳定性:方法:通过核磁共振定量分析进行校准,以确保准确性。在 96 孔板中使用氚代 C4 内标和液液萃取法分析 C4。验证遵循 2018 年食品与药物管理局指南。为评估 C4 的稳定性,健康志愿者(n=12)每人捐献 8 份空腹样本。样品在20 °C下孵育长达72小时,在C4分析前的不同时间点取回、离心、等分并冷冻保存:结果:C4 方法具有出色的分析性能,通过了所有验证标准。该方法准确、精确、无基质效应和干扰。样品延迟分离 72 小时后,C4 浓度从基线逐渐下降,降幅达 14%。然而,在长达 12 小时的时间里,这种变化并不明显:我们提出了一种分析血清 C4 的可靠方法,为 BAD 调查提供了一种替代 75SeHCAT 的便捷方法。我们发现未分离血液中的 C4 会随着时间的推移而下降;然而,12 小时后的平均变化是
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development and validation of a novel 7α-hydroxy-4-cholesten-3-one (C4) liquid chromatography tandem mass spectrometry method and its utility to assess pre-analytical stability.

Objectives: 7α-Hydroxy-4-cholesten-3-one (C4) is the common intermediary of both primary bile acids. C4 is recommended by the British Society of Gastroenterology for the investigation of bile acid diarrhoea (BAD) in patients with chronic diarrhoea. This project aimed to develop and validate an assay to quantitate C4 in serum and assess the stability of C4 in unseparated blood.

Methods: Accuracy was underpinned by calibrating to quantitative nuclear magnetic resonance analysis. C4 was analysed in a 96-well plate format with a deuterated C4 internal standard and liquid-liquid extraction. Validation followed the 2018 Food and Drug Administration guidelines. To assess C4 stability, healthy volunteers (n=12) donated 8 fasted samples each. Samples were incubated at 20 °C for up to 72 h and retrieved, centrifuged, aliquoted and frozen for storage at different time points prior to C4 analysis.

Results: The C4 method demonstrated excellent analytical performance and passed all validation criteria. The method was found to be accurate, precise, free from matrix effects and interference. After 72 h of delayed sample separation, C4 concentration gradually declined by up to 14 % from baseline. However, the change was not significant for up to 12 h.

Conclusions: We present a robust method of analysing serum C4, offering a convenient alternative to 75SeHCAT for BAD investigation. C4 was found to decline in unseparated blood over time; however, after 12 h the mean change was <5 % from baseline. Our results suggest C4 is suitable for collection from both primary and secondary care prior to gastroenterology referral.

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来源期刊
Clinical chemistry and laboratory medicine
Clinical chemistry and laboratory medicine 医学-医学实验技术
CiteScore
11.30
自引率
16.20%
发文量
306
审稿时长
3 months
期刊介绍: Clinical Chemistry and Laboratory Medicine (CCLM) publishes articles on novel teaching and training methods applicable to laboratory medicine. CCLM welcomes contributions on the progress in fundamental and applied research and cutting-edge clinical laboratory medicine. It is one of the leading journals in the field, with an impact factor over 3. CCLM is issued monthly, and it is published in print and electronically. CCLM is the official journal of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) and publishes regularly EFLM recommendations and news. CCLM is the official journal of the National Societies from Austria (ÖGLMKC); Belgium (RBSLM); Germany (DGKL); Hungary (MLDT); Ireland (ACBI); Italy (SIBioC); Portugal (SPML); and Slovenia (SZKK); and it is affiliated to AACB (Australia) and SFBC (France). Topics: - clinical biochemistry - clinical genomics and molecular biology - clinical haematology and coagulation - clinical immunology and autoimmunity - clinical microbiology - drug monitoring and analysis - evaluation of diagnostic biomarkers - disease-oriented topics (cardiovascular disease, cancer diagnostics, diabetes) - new reagents, instrumentation and technologies - new methodologies - reference materials and methods - reference values and decision limits - quality and safety in laboratory medicine - translational laboratory medicine - clinical metrology Follow @cclm_degruyter on Twitter!
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