超高效液相色谱法测定血浆中毒死蜱及其主要代谢产物

Detecting chlorpyrifos and its metabolitein plasma by UPLC

目的建立血浆中毒死蜱（CPF）及其主要代谢物3,5,6-三氯-2-吡啶醇（TCP）的超高效液相色谱（UPLC）测定方法。方法取0.50 m L待测血浆经乙酸乙酯萃取,氮气吹干,乙腈定容,采用C18柱分离,以乙腈∶水为流动相梯度洗脱,在二极管阵列检测器检测波长为298 nm（0.0~2.0 min）和289 nm（2.0~5.0 min）的紫外光谱下,经UPLC仪测定。结果 CPF和TCP在质量浓度为1.00~50.00 mg/L时线性关系良好,相关系数均为0.999 9,检出限均为0.30 mg/L,定量限均为1.00 mg/L。加标回收率分别为87.20%~103.08%和86.20%~99.28%,批内相对标准偏差（RSD）分别为5.28%~6.17%和2.32%~4.43%,批间RSD分别为6.62%~7.53%和3.55%~5.24%。样品在4℃冰箱内可保存7 d。结论本方法准确可靠,简便实用,可用于测定血浆中CPF和TCP。

Objective To establish a simple and sensitive method for simultaneous detection of chlorpyrifos（ CPF）,and its metabolite 3,5,6-trichloro-2-pyridinol（ TCP） in plasma samples by ultra performance liquid chromatography（ UPLC）.Methods The 0. 5 m L of blood sample was extracted by ethyl acetate,then separated by C18 column using acetonitrile /water as a mobile phase and detected by diode-array detector under the ultraviolet spectrum of 298 nm（ 0. 0-2. 0 min） and289 nm（ 2. 0-5. 0 min） through UPLC. Results The method showed that a good linear range was 1. 00-50. 00 mg / L of both CPF and TCP,and the correlation coefficients was 0. 999 9. The limit of detection was 0. 30 mg / L and the lower limits of quantitation was 1. 00 mg / L of both CPF and TCP. The recovery rates of CPF and TCP were 87. 20 %-103. 08 %and 86. 20 %-99. 28 %,respectively. The within-run relative standard deviation（ RSD） of CPF and TCP was 5. 28 %-6. 17 % and 2. 32 %-4. 43 %,and the between-run RSD was 6. 62 %-7. 53 % and 3. 55 %-5. 24 %. Samples can be kept in4 degrees Celsius refrigerator for 7 days. Conclusions The proposed method is simple,reliable and sensitive. It can be applied for simultaneous detection of CPF and its metabolite TCP in plasma samples.