Improved real-world UHPLC-MS/MS iohexol analysis; thawed samples, improved calibration curve, and additional matrices.

IF 1.8 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Bioanalysis Pub Date : 2025-03-01 Epub Date: 2025-02-07 DOI:10.1080/17576180.2025.2462429

A Mireille A Wessels, Lenneke A T Junier, Daan Touw, Jasper Stevens

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引用次数: 0

Abstract

Background: Plasma clearance of iohexol is used to measure glomerular filtration rate, for which a UHPLC-MS/MS analytical method was previously developed. In real-world conditions, samples may be thawed on arrival and sampled in unvalidated matrices, prompting the need for an improved validation. We aim to improve the method for iohexol determination in plasma with enhanced stability testing, optimized calibration curves, and partial validation in additional matrices.

Methods: Stability testing was conducted up to 9 weeks at room temperature, 4°C, 37°C, and at 37°C with daily two-hour exposure to 55°C. Quintuplicate QC samples were analyzed on 3 days, comparing results from eight-point and two-point calibration curves. Matrix comparison was performed on quintuplicate QC samples in serum, heparin plasma, urine, EDTA whole blood, and heparin whole blood.

Results: The method improvements were all compliant with the requirements for bioanalytical methods issued by the US FDA and European Medicines Agency.

Conclusion: Human EDTA plasma samples can be stored up to 9 weeks at room temperature, 4°C, 37°C, and 37°C with 55°C daily temperature spikes. The samples can be analyzed using a two-point calibration curve and are partial validated for serum-, heparin plasma-, urine-, EDTA whole blood-, and lithium whole blood iohexol samples.

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改进的UHPLC-MS/MS碘己醇分析方法解冻的样品，改进的校准曲线，和额外的基质。

背景：血浆碘己醇清除率用于测定肾小球滤过率，此前开发了UHPLC-MS/MS分析方法。在实际条件下，样品可能在到达时解冻，并在未经验证的基质中取样，这促使需要改进验证。我们的目的是通过增强稳定性测试、优化校准曲线和在附加基质上的部分验证来改进血浆中碘己醇的测定方法。方法：稳定性试验在室温、4°C、37°C和37°C下进行长达9周，每天暴露于55°C下2小时。在3天内分析5个QC样品，比较8点和2点校准曲线的结果。对血清、肝素血浆、尿液、EDTA全血、肝素全血中的5份QC样品进行基质比较。结果：改进后的方法均符合美国FDA和欧洲药品管理局颁布的生物分析方法要求。结论：人EDTA血浆样品可在室温、4°C、37°C和37°C条件下保存9周，每日温度峰值为55°C。样品可以使用两点校准曲线进行分析，并对血清-、肝素-、血浆-、尿-、EDTA全血-和锂全血碘己醇样品进行部分验证。

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

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.