Kuijpers E A, den Hartigh J, Savelkoul T J, de Wolff F A
Department of Clinical Chemistry, Pharmacy and Toxicology, Leiden University Hospital, The Netherlands.
J Anal Toxicol. 1995 Nov-Dec;19(7):557-62. doi: 10.1093/jat/19.7.557.
A high-performance liquid chromatographic method with ultraviolet (UV) and fluorescence detection was developed for the analysis of one indandione and four hydroxycoumarin anticoagulant rodenticides in human serum. The superwarfarin rodenticides, chlorophacinone, bromadiolone, difenacoum, brodifacoum, and difethialone, can be identified and quantitated simultaneously with this method. After adding a buffer (pH 5.5), the anticoagulants were extracted from serum with chloroform-acetone. The organic phase was separated and evaporated to dryness, and the residue was subjected to chromatographic analysis. The anticoagulants were separated by reversed-phase chromatography and detected by UV absorption at 285 nm and by fluorescence at an excitation wavelength of 265 nm and an emission wavelength of 400 nm. Extraction efficiencies from 55 to 131% were obtained. The within-run precision ranged from 2.0 to 7.1% for UV detection and from 0.0 to 4.8% for fluorescence detection. Between-run precision ranged from 1.3 to 16.0% for UV detection and from 1.8 to 9.0% for fluorescence detection. The anticoagulants can be quantitated at serum concentrations down to 3-12 ng/mL for fluorescence detection and down to 20-75 ng/mL for UV detection. No interferences were observed with the related compounds warfarin and vitamin K1.
建立了一种采用紫外（UV）和荧光检测的高效液相色谱法，用于分析人血清中的一种茚二酮和四种羟基香豆素类抗凝血杀鼠剂。使用该方法可同时鉴定和定量超级杀鼠灵类杀鼠剂氯鼠酮、溴敌隆、敌鼠、溴鼠灵和二氟隆。加入缓冲液（pH 5.5）后，用氯仿 - 丙酮从血清中提取抗凝血剂。分离有机相并蒸发至干，残渣进行色谱分析。抗凝血剂通过反相色谱分离，在285 nm处通过紫外吸收以及在激发波长265 nm和发射波长400 nm处通过荧光进行检测。提取效率为55%至131%。批内精密度对于紫外检测为2.0%至7.1%，对于荧光检测为0.0%至4.8%。批间精密度对于紫外检测为1.3%至16.0%，对于荧光检测为1.8%至9.0%。对于荧光检测，抗凝血剂在血清浓度低至3 - 12 ng/mL时可进行定量，对于紫外检测，低至20 - 75 ng/mL时可进行定量。未观察到与相关化合物华法林和维生素K1的干扰。
