鳗鲡血浆与肌肉中乙胺嘧啶检测方法的建立

Establishment of Detection method of Pyrimethamine in Plasma and Mussels of Eel

通过萃取回收和色谱条件优化,建立了运用高效液相色谱法(HPLC)检测鳗鲡血浆和肌肉中乙胺嘧啶的方法。结果显示:在柱温30℃、检测波长236 nm、流动相乙腈、水、乙酸体积比为15∶85∶0.25和流速1.0 mL/min的色谱条件下,在设置鳗鲡血浆及肌肉中乙胺嘧啶浓度分别在0～5.0μg/mL与0～2.0μg/g范围内,相关曲线方程分别为y=766.68x+9.2511与y=853.96x+8.0956,相关系数(r)分别为0.9998和0.9999。在萃取回收中,向血浆和肌肉样品中分别加入适量无水硫酸钠,再分别用乙腈、三氯甲烷体积比为10∶1和5∶5的混合液萃取两次,回收率均达80%以上,日间、日内变异系数均低于3%,最低检测限可分别达0.005μg/mL和0.005μg/g。

The method of testing of pyrimethamine（PYR） in plasma and muscle of eel was established by optimized extraction parameters and analysis conditions for high-performance liquid chromatography（HPLC）. The results indicated： when the column temperature was 30 ℃, the detecting wavelength was 236 nm, the mobile phase was consisted of acetnonitrile- water-acetic acid（ 15： 85：0.25 v/v/v） and the flow rate was 1.0 mL/min, the equations within 0 - 5.0 μg/mL and 0-2. 0 μg/g of PYR in plasma and muscle of eel were y = 766. 68x ＋ 9. 2511 and y = 853.96x ＋ 8. 0956, correlation coefficients was 0. 9998 and 0. 9999 respectively; after the suitable quality of sodium sulfate anhydrous was firstly added to both plasma and muscle and then two times extractions were respectively performed by mixed solution of acetonitrile-trichlormethane（10：1 v/v）and acetonitrile-trichlormethane（5：5 v/v）, the recover rate were both over 80%, the coefficients of variation were less than 3% and the lowest detective limit was 0. 005 μg/mL（μg/g）.