Department of Pathology and Microbiology, School of Medicine, University of Zambia, Lusaka 10101, Zambia.
Centre for Human Metabolomics, North-West University, Potchefstroom 2531, South Africa.
Molecules. 2023 Aug 7;28(15):5927. doi: 10.3390/molecules28155927.
The aim of this study was to improve the extraction method for urinary organic acids by miniaturizing and automating the process. Currently, manual extraction methods are commonly used, which can be time-consuming and lead to variations in test results. To address these issues, we reassessed and miniaturized the in-house extraction method, reducing the number of steps and the sample-to-solvent volumes required. The evaluated miniaturized method was translated into an automated extraction procedure on a MicroLab (ML) Star (Hamilton Technologies) liquid handler. This was then validated using samples obtained from the ERNDIM External Quality Assurance program. The organic acid extraction method was successfully miniaturized and automated using the Autosampler robot. The linear range for most of the thirteen standard analytes fell between 0 to 300 mg/L in spiked synthetic urine, with low (50 mg/L), medium (100 mg/L), and high (500 mg/L) levels. The correlation coefficient (r) for most analytes was >0.99, indicating a strong relationship between the measured values. Furthermore, the automated extraction method demonstrated acceptable precision, as most organic acids had coefficients of variation (CVs) below 20%. In conclusion, the automated extraction method provided comparable or even superior results compared to the current in-house method. It has the potential to reduce solvent volumes used during extraction, increase sample throughput, and minimize variability and random errors in routine diagnostic settings.
本研究旨在通过小型化和自动化处理过程来改进尿液有机酸的提取方法。目前，通常采用手动提取方法，但该方法耗时且可能导致测试结果出现差异。为了解决这些问题，我们重新评估并小型化了内部提取方法，减少了所需的步骤和样品与溶剂的体积。评估后的小型化方法被转化为 Hamilton Technologies 公司的 MicroLab（ML）Star 上的自动化提取程序。然后使用 ERNDIM 外部质量保证计划获得的样本对其进行验证。有机酸提取方法使用 Autosampler 机器人成功地实现了小型化和自动化。在加标合成尿液中，大多数标准分析物的 13 种标准分析物的线性范围在 0 至 300mg/L 之间，低（50mg/L）、中（100mg/L）和高（500mg/L）水平。大多数分析物的相关系数（r）大于 0.99，表明测量值之间存在很强的相关性。此外，自动化提取方法的精密度可接受，因为大多数有机酸的变异系数（CV）低于 20%。总之，与当前的内部方法相比，自动化提取方法提供了相当甚至更好的结果。它有可能减少提取过程中使用的溶剂体积，增加样品通量，并最大限度地减少常规诊断环境中的变异性和随机误差。
