Metronidazole is a chromatographic interference on a routine HPLC-UV assay for thiopurine metabolites.

IF 1 4区医学 Q3 MEDICAL LABORATORY TECHNOLOGY

Annals of Clinical Biochemistry Pub Date : 2025-02-21 DOI:10.1177/00045632251319917

Andrew A Wood, Sophie Rothwell Mason, Anna F Robson

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引用次数: 0

Abstract

Background: Thiopurine metabolites, 6-thioguanine (6TG) and 6-methylmercaptopurine (6MMP), are monitored to aid therapeutic management of thiopurine drugs. At Manchester University NHS Foundation Trust (MFT), thiopurine metabolites are measured by high performance liquid chromatography with ultraviolet detection (HPLC-UV). Whole blood samples are lysed and subjected to hydrolysis with derivatisation of 6MMP before HPLC-UV detection at 304 nm for the 6MMP-derivative and 342 nm for 6TG. For some samples, 6MMP cannot be reported due to a chromatographic interference at 304 nm co-eluting with the 6MMP peak. An investigation was performed to identify the interfering compound.

Methods: Patient medication histories were examined to identify candidate compounds for the interference. Candidate compounds were spiked into blood at supraphysiological concentrations and tested on the assay.

Results: Metronidazole was identified as being prescribed to all patients whose samples demonstrated the interference. Metronidazole and its metabolite, hydroxymetronidazole, were spiked into blood. HPLC-UV analysis of spiked blood demonstrated similar UV absorbance patterns to those seen in patient samples with the interference. Hydroxymetronidazole co-eluted with 6MMP causing interference in the measurement.

Conclusion: Metronidazole and its major metabolite can interfere with 6MMP measurement by HPLC-UV analysis at 304 nm.

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甲硝唑是常规HPLC-UV测定硫嘌呤代谢物的色谱干扰物。

监测硫嘌呤代谢物6-硫鸟嘌呤（6TG）和6-甲基巯基嘌呤（6MMP），以帮助硫嘌呤药物的治疗管理。在曼彻斯特大学NHS基金会信托（MFT），硫嘌呤代谢物通过高效液相色谱与紫外检测（HPLC-UV）进行测量。全血样品被裂解并进行6MMP衍生化水解，然后在304nm处进行6MMP衍生和342nm处进行6TG的HPLC-UV检测。对于某些样品，由于在304nm处与6MMP峰共洗脱的色谱干扰，无法报告6MMP。对干扰物进行了鉴定。方法通过对患者用药史的调查，确定干扰药物的候选化合物。候选化合物以超生理浓度加入血液，并在分析中进行测试。结果甲硝唑对所有样品有干扰的患者均有处方。将甲硝唑及其代谢物羟基甲硝唑加入血液中。高效液相色谱-紫外分析的加标血液显示类似的紫外线吸收模式，这些在患者样品中看到的干扰。羟甲硝唑与6MMP共洗脱导致测量干扰。结论甲硝唑及其主要代谢物可干扰304 nm HPLC-UV测定6MMP。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Annals of Clinical Biochemistry Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry

CiteScore

5.20

自引率

4.50%

发文量

期刊介绍： Annals of Clinical Biochemistry is the fully peer reviewed international journal of the Association for Clinical Biochemistry and Laboratory Medicine. Annals of Clinical Biochemistry accepts papers that contribute to knowledge in all fields of laboratory medicine, especially those pertaining to the understanding, diagnosis and treatment of human disease. It publishes papers on clinical biochemistry, clinical audit, metabolic medicine, immunology, genetics, biotechnology, haematology, microbiology, computing and management where they have both biochemical and clinical relevance. Papers describing evaluation or implementation of commercial reagent kits or the performance of new analysers require substantial original information. Unless of exceptional interest and novelty, studies dealing with the redox status in various diseases are not generally considered within the journal''s scope. Studies documenting the association of single nucleotide polymorphisms (SNPs) with particular phenotypes will not normally be considered, given the greater strength of genome wide association studies (GWAS). Research undertaken in non-human animals will not be considered for publication in the Annals. Annals of Clinical Biochemistry is also the official journal of NVKC (de Nederlandse Vereniging voor Klinische Chemie) and JSCC (Japan Society of Clinical Chemistry).