Mudiam Mohana Krishna Reddy, Singh Mahendra Pratap, Chowdhuri Debapratim Kar, Murthy Ramesh Chandra
Indian Institute of Toxicology Research (Council of Scientific & Industrial Research), Analytical Chemistry Section, Mahatma Gandhi Marg, Lucknow, India.
J AOAC Int. 2010 Sep-Oct;93(5):1595-9.
A simple, rapid, and solvent-free method for quantitative determination of benzene, toluene, and Xylene in exposed Drosophila larvae was developed using headspace solid-phase microextraction (HS-SPME) coupled to GC/MS. Larvae fed on standard Drosophila food mixed with benzene, toluene, and Xylene for 48 h were homogenized in Milli-Q water. Extraction of benzene, toluene, and Xylene was performed at 65 degrees C for 30 min on the SPME fiber (silica-fused). Subsequently, the fiber was desorbed in the GC injection port, followed by GC/MS analysis in the selected-ion monitoring mode. An external calibration curve was used for the quantification of benzene, toluene, and Xylene in the exposed organism. Recoveries were in the range of 78-82% (intraday) and 76-81% (interday) in larvae, and 91-96% (intraday) and 87-92% (interday) in the diet. LOD with an S/N of 3:1 and LOQ with an S/N of 10:1 were in the range of 0.01-0.023 and 0.034-0.077 microg/L, respectively. Percent RSD values for benzene, toluene, and Xylene were in the range of 0.50-0.81 (intraday) and 0.89-1.23 (interday) for retention time, and 2.16--3.85 (intraday) and 2.99-4.95 (interday) for peak concentration, showing good repeatability. This method was sensitive enough to quantitate benzene, toluene, and Xylene in small exposed organisms like Drosophila larvae. The SPME/GC/MS method developed may have wider applications in various in vivo toxicological studies.
开发了一种简单、快速且无溶剂的方法，用于定量测定暴露的果蝇幼虫中的苯、甲苯和二甲苯，该方法采用顶空固相微萃取（HS-SPME）与气相色谱/质谱联用。以混合有苯、甲苯和二甲苯的标准果蝇食物喂养幼虫48小时后，将其在超纯水（Milli-Q水）中匀浆。在65℃下，使用SPME纤维（石英纤维）对苯、甲苯和二甲苯进行30分钟的萃取。随后，将纤维在气相色谱进样口中解吸，然后在选择离子监测模式下进行气相色谱/质谱分析。采用外标曲线对暴露生物体中的苯、甲苯和二甲苯进行定量。幼虫体内苯、甲苯和二甲苯的日内回收率在78 - 82%之间，日间回收率在76 - 81%之间；食物中的日内回收率在91 - 96%之间，日间回收率在87 - 92%之间。信噪比为3:1时的检测限（LOD）在0.01 - 0.023 μg/L范围内，信噪比为10:1时的定量限（LOQ）在0.034 - 0.077 μg/L范围内。苯、甲苯和二甲苯保留时间的日内相对标准偏差（RSD）百分比在0.50 - 0.81之间，日间在0.89 - 1.23之间；峰浓度的日内RSD百分比在2.16 - 3.85之间，日间在2.99 - 4.95之间，显示出良好的重复性。该方法灵敏度足以对果蝇幼虫等小型暴露生物体中的苯、甲苯和二甲苯进行定量。所开发的SPME/GC/MS方法可能在各种体内毒理学研究中有更广泛的应用。
