High throughput bioanalysis of serum 7a-hydroxy-4-cholesten-3-one (C4) using LC-MS/MS: Devising an end-to-end single vial solution for a sample limited application

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis Pub Date : 2024-11-26 DOI:10.1016/j.jpba.2024.116581

Soumya Kandi , Sarah Blink Polakow , John P. Savaryn , Kenneth Ruterbories , Mary Saltarelli , Gary J. Jenkins , Qin C. Ji

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引用次数: 0

Abstract

Crohn’s disease (CD) is characterized by chronic ileal/ileocolonic inflammation, and in some cases, can result in bile acid malabsorption (BAM) and subsequent bile acid diarrhea (BAD). Although BAD is common in CD, diagnosis is difficult. In patients with CD who had ileal resection (IR), elevated serum 7a-hydroxy-4-cholesten-3-one (C4), a cholesterol-derived stable intermediate in bile acid synthesis, is associated with diarrhea attributable to BAM and therefore, may have diagnostic utility. The previously existing validated methodology to measure C4 in human serum required 100 μL with an analytical range of 0.5–100 ng/mL, making it incompatible with the clinical trial sample set we had available for analysis due to limited serum volume and an extended assay range requirement (up-to 1 μg/mL). We present here a simplified, end-to-end single vial approach performed in a 96-well format for clinical sample C4 analysis for application in sample limited settings.

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使用LC-MS/MS对血清7a-羟基-4-胆甾醇-3-one （C4）进行高通量生物分析：为样品有限的应用设计端到端单瓶溶液。

克罗恩病（CD）的特点是慢性回肠/回肠结肠炎症，在某些情况下，可导致胆汁酸吸收不良（BAM）和随后的胆汁酸腹泻（BAD）。虽然BAD在乳糜泻中很常见，但诊断却很困难。在行回肠切除术（IR）的CD患者中，血清7a-羟基-4-胆固醇-3- 1 （C4）升高与BAM引起的腹泻有关，C4是胆汁酸合成中胆固醇衍生的稳定中间体，因此可能具有诊断价值。先前已验证的测定人血清中C4的方法需要100 μL，分析范围为0.5-100 ng/mL，由于有限的血清体积和扩展的分析范围要求（高达1 μg/mL），使其与我们可用的临床试验样品集不兼容。我们在这里提出了一种简化的端到端单瓶方法，采用96孔格式，用于临床样品C4分析，适用于样品有限的环境。

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来源期刊

Journal of pharmaceutical and biomedical analysis 医学-分析化学

CiteScore

6.70

自引率

5.90%

发文量

588

审稿时长

37 days

期刊介绍： This journal is an international medium directed towards the needs of academic, clinical, government and industrial analysis by publishing original research reports and critical reviews on pharmaceutical and biomedical analysis. It covers the interdisciplinary aspects of analysis in the pharmaceutical, biomedical and clinical sciences, including developments in analytical methodology, instrumentation, computation and interpretation. Submissions on novel applications focusing on drug purity and stability studies, pharmacokinetics, therapeutic monitoring, metabolic profiling; drug-related aspects of analytical biochemistry and forensic toxicology; quality assurance in the pharmaceutical industry are also welcome. Studies from areas of well established and poorly selective methods, such as UV-VIS spectrophotometry (including derivative and multi-wavelength measurements), basic electroanalytical (potentiometric, polarographic and voltammetric) methods, fluorimetry, flow-injection analysis, etc. are accepted for publication in exceptional cases only, if a unique and substantial advantage over presently known systems is demonstrated. The same applies to the assay of simple drug formulations by any kind of methods and the determination of drugs in biological samples based merely on spiked samples. Drug purity/stability studies should contain information on the structure elucidation of the impurities/degradants.