Department of Chemistry, Science and Research Branch, Islamic Azad University, Kerman, Iran.
Department of Chemistry, Kerman Branch, Islamic Azad University, Kerman, Iran.
Arh Hig Rada Toksikol. 2020 Mar 1;71(1):48-55. doi: 10.2478/aiht-2020-71-3292.
Diazinon poisoning is an important issue in occupational, clinical, and forensic toxicology. While sensitive and specific enough to analyse diazinon in biological samples, current methods are time-consuming and too expensive for routine analysis. The aim of this study was therefore to design and validate a simple dispersive liquid-liquid microextraction (DLLME) for the preparation of urine samples to be analysed for diazinon with high performance liquid chromatography with diode-array detector (HPLC-DAD) to establish diazinon exposure and poisoning. To do that, we first identified critical parameters (type and volume of extraction and disperser solvents, pH, surfactant, and salt concentrations) in preliminary experiments and then used central composite design to determine the best experimental conditions for DLLME-HPLC-DAD. For DLLME they were 800 µL of methanol (disperser solvent) and 310 µL of toluene (extraction solvent) injected to the urine sample rapidly via a syringe. The sample was injected into a HPLC-DAD (C18 column, 250×4.6 mm, 5 μm), and the mobile phase was a mixture of acetonitrile and buffer (63:37 v/v, pH 3.2; flow rate: 1 mL/ min). Standard calibration curves for diazinon were linear with the concentration range of 0.5-4 µg/mL, yielding a regression equation Y=0.254X+0.006 with a correlation coefficient of 0.993. The limit of detection and limit of quantification for diazinon were 0.15 µg/mL and 0.45 µg/mL, respectively. The proposed method was accurate, precise, sensitive, and linear over a wide range of diazinon concentrations in urine samples. This method can be employed for diazinon analysis in routine clinical and forensic toxicology settings.
敌百虫中毒是职业、临床和法医毒理学中的一个重要问题。虽然现有的分析方法对于生物样本中的敌百虫检测足够灵敏和特异，但对于常规分析来说，这些方法既耗时又昂贵。因此，本研究旨在设计并验证一种简单的分散液-液微萃取（DLLME）方法，用于制备尿液样品，以便使用高效液相色谱-二极管阵列检测器（HPLC-DAD）进行分析，从而建立敌百虫暴露和中毒的检测方法。为此，我们首先在初步实验中确定了关键参数（萃取和分散溶剂的类型和体积、pH 值、表面活性剂和盐浓度），然后使用中心复合设计确定 DLLME-HPLC-DAD 的最佳实验条件。对于 DLLME，我们快速地将 800µL 甲醇（分散溶剂）和 310µL 甲苯（萃取溶剂）注入尿液样品中，然后将样品注入 HPLC-DAD（C18 柱，250×4.6mm，5µm），流动相为乙腈和缓冲液的混合物（63:37v/v，pH3.2；流速：1mL/min）。敌百虫的标准校准曲线在 0.5-4µg/mL 的浓度范围内呈线性，回归方程为 Y=0.254X+0.006，相关系数为 0.993。敌百虫的检出限和定量限分别为 0.15µg/mL 和 0.45µg/mL。该方法在尿液样品中敌百虫浓度较宽的范围内具有准确性、精密度、灵敏度和线性。该方法可用于常规临床和法医毒理学中敌百虫的分析。
