Bidny Sergei, Gago Kim, David Mark, Duong Thanh, Albertyn Desdemona, Gunja Naren
Forensic and Analytical Science Service, NSW Health Pathology, Sydney, Australia
Forensic and Analytical Science Service, NSW Health Pathology, Sydney, Australia.
J Anal Toxicol. 2015 Apr;39(3):219-24. doi: 10.1093/jat/bku175. Epub 2015 Jan 16.
A rapid, highly sensitive and specific analytical method for the extraction, identification and quantification of nine rodenticides from whole blood has been developed and validated. Commercially available rodenticides in Australia include coumatetralyl, warfarin, brodifacoum, bromadiolone, difenacoum, flocoumafen, difethialone, diphacinone and chlorophacinone. A Waters ACQUITY UPLC TQD system operating in multiple reaction monitoring mode was used to conduct the analysis. Two different ionization techniques, ES+ and ES-, were examined to achieve optimal sensitivity and selectivity resulting in detection by MS-MS using electrospray ionization in positive mode for difenacoum and brodifacoum and in negative mode for all other analytes. All analytes were extracted from 200 µL of whole blood with ethylacetate and separated on a Waters ACQUITY UPLC BEH-C18 column using gradient elution. Ammonium acetate (10 mM, pH 7.5) and methanol were used as mobile phases with a total run time of 8 min. Recoveries were between 70 and 105% with limits of detection ranging from 0.5 to 1 ng/mL. The limit of quantitation was 2 ng/mL for all analytes. Calibration curves were linear within the range 2-200 ng/mL for all analytes with the coefficient of determination ≥0.98. The application of the proposed method using liquid-liquid extraction in a series of clinical investigations and forensic toxicological analyses was successful.
已开发并验证了一种用于从全血中提取、鉴定和定量九种灭鼠剂的快速、高灵敏度和特异性分析方法。澳大利亚市售的灭鼠剂包括杀鼠灵、华法林、溴敌隆、溴鼠灵、敌鼠、氟鼠灵、二氟杀鼠酮、敌鼠钠盐和氯鼠酮。使用在多反应监测模式下运行的沃特世ACQUITY UPLC TQD系统进行分析。研究了两种不同的电离技术，即ES+和ES-，以实现最佳灵敏度和选择性，结果采用电喷雾电离在正模式下对敌鼠和溴敌隆进行MS-MS检测，对所有其他分析物在负模式下进行检测。所有分析物均用乙酸乙酯从200 μL全血中提取，并在沃特世ACQUITY UPLC BEH-C18柱上使用梯度洗脱进行分离。乙酸铵（10 mM，pH 7.5）和甲醇用作流动相，总运行时间为8分钟。回收率在70%至105%之间，检测限为0.5至1 ng/mL。所有分析物的定量限均为2 ng/mL。所有分析物在2-200 ng/mL范围内校准曲线呈线性，决定系数≥0.98。所提出的液-液萃取方法在一系列临床研究和法医毒理学分析中的应用是成功的。
