Sarani Behnaz, Rahmani Mashaallah, Abbasian Ahmad Reza
Department of Chemistry, Faculty of Sciences, University of Sistan and Baluchestan, Zahedan, Iran.
Department of Materials Engineering, Faculty of Engineering, University of Sistan and Baluchestan, Zahedan, Iran.
Sci Rep. 2025 Aug 12;15(1):29585. doi: 10.1038/s41598-025-14975-0.
This study employed a quick, efficient, sensitive, and easy-to-use micro-extraction process to monitor biomarker hippuric acid (BIHA) in urine samples as a bioindicator of exposure to toluene with an HPLC-DAD instrument. The independent variables for the micro-extraction process of hippuric acid were examined, while the response from the HA monitoring acted as the dependent variable. Initial screening and parameter optimization were performed using the multivariate models. The desirability function aimed to maximize the extraction recovery of hippuric acid while minimizing solvent volume and extraction time. The best conditions identified were: 152 µL of extractant solvent, extraction time 253 s (4.2 min), a salt amount of 3.9%W/V, and pH = 2.7 with Desirability = 1. Achieving Desirability = 1 reflects a perfect compromise of all goals. The suggested model showed strong validation, evidenced by adjusted R² = 0.9888, high recovery rates, low error in confirmation runs (1.37-2.09%), and consistent predicted values. The model demonstrated satisfactory recovery rates, emphasizing its potential for analyzing urine samples as an indicator of toluene exposure. The detection limit was established at 0.15 ng mL for hippuric acid, with a calibration curve linear of 1 to 900 ng mL. Assessments of samples conducted with this model established its validity and resulted in adequate efficiency. The precision assessment of the technique (as %RSD) was found to be in the range of 2.11-4.25%.
本研究采用了一种快速、高效、灵敏且易于使用的微萃取方法，使用高效液相色谱 - 二极管阵列检测器（HPLC - DAD）仪器监测尿液样本中的生物标志物马尿酸（BIHA），作为甲苯暴露的生物指标。研究考察了马尿酸微萃取过程中的自变量，而马尿酸监测的响应作为因变量。使用多变量模型进行了初步筛选和参数优化。合意函数旨在最大化马尿酸的萃取回收率，同时最小化溶剂体积和萃取时间。确定的最佳条件为：萃取剂溶剂152微升，萃取时间253秒（4.2分钟），盐含量3.9%W/V，pH = 2.7，合意度 = 1。达到合意度 = 1反映了所有目标的完美折衷。所建议的模型显示出强大的有效性，调整后的R² = 0.9888、回收率高、验证运行中的误差低（1.37 - 2.09%）以及预测值一致均证明了这一点。该模型显示出令人满意的回收率，强调了其作为甲苯暴露指标分析尿液样本的潜力。马尿酸的检测限确定为0.15纳克/毫升，校准曲线在1至900纳克/毫升范围内呈线性。用该模型对样本进行的评估确定了其有效性并具有足够的效率。该技术的精密度评估（以%RSD计）在2.11 - 4.25%范围内。
