高效液相色谱法测定全血中5种抗凝血类灭鼠剂

Determination of five anticoagulant rodenticides in whole blood by high-performance liquid chromatography

目的建立同时测定全血中溴鼠隆、溴敌隆、氟鼠灵、杀鼠灵和鼠得克5种灭鼠剂的高效液相色谱测定方法。方法取0.5 m L全血样品经2.0 mL乙酸乙酯萃取后,采用Diamonsil C18（250.0 mm×4.6 mm×5.0μm）色谱柱分离,以浓度为20 mmol/L的乙酸铵-乙酸缓冲液（p H值=5.5）∶甲醇（2∶8,V/V）为流动相,在二极管阵列检测器检测波长为310 nm的紫外光谱下经高效液相色谱仪测定。结果全血中溴鼠隆、溴敌隆、氟鼠灵、杀鼠灵和鼠得克在质量浓度为0.50～10.00 mg/L线性关系良好,相关系数均大于0.999 00,检出限分别为0.08、0.06、0.09、0.04和0.10 mg/L,定量下限分别为0.79、0.58、0.92、0.45和0.96 mg/L。加标回收率为98.40%～104.00%。批内相对标准偏差为0.61%～7.84%,批间相对标准偏差为1.10%～9.62%。样品在4℃冰箱内可保存14 d。结论本方法操作简单、分离效果好、灵敏度高,具有较好的精密度和准确度,适用于灭鼠剂中毒患者全血样品的检测。

Objective To establish a method for simultaneous detection of 5 anticoagulant rodenticides including brodifacoum, bromadiolone, flocoumafen, warfarin and difenacoum in whole blood by high-performance liquid chromatogarphy. Methods The 0. 5 m L of blood sample was extracted by 2. 0 m L ethyl acetate,then separated by Diamonsil C18 column（ 250. 0 mm × 4. 6 mm × 5. 0 μm） using acid-ammonium acetate（ 20. 0 mmol / L,p H = 5. 5） /methyl alcohol（ 2∶ 8,V / V） as a mobile phase and detected by diode-array detector under the ultraviolet spectrum of 310 nm through high-performance liquid chromatogarphy. Results The good linear range of the 5 anticoagulant rodenticides was0. 50-10. 00 mg / L,and the correlation coefficients were〉 0. 999 00. The detection limits of brodifacoum,bromadiolone,flocoumafen,warfarin and difenacoum were 0. 08,0. 06,0. 09,0. 04 and 0. 10 mg / L and the lower limits of quantitation were 0. 79,0. 58,0. 92,0. 45 and 0. 96 mg / L,respectively. The recovery rate was 98. 40%-104. 00%. The within-run relative standard deviation（ RSD） was 0. 61%-7. 84%,and the between-run RSD was 1. 10%-9. 62%. The samples can be stored for 14 days in the refrigerator at 4 ℃. Conclusion The method has advantages of simple operation,good separating effect,high sensitivity,precision and accuracy,which is suitable for detection of whole blood samples in rodenticide poisoning patients.