猪肌肉与肝脏中卡拉洛尔高效液相色谱检测方法的研究

Determination of Carazolol in Swine Muscle and Liver by High-performance Liquid Chromatography

建立猪肌肉与肝脏中卡拉洛尔残留的高效液相色谱荧光检测法。样品经乙腈提取,提取液经正己烷两次除脂后浓缩净化,用乙腈-水(2∶8,V/V)溶解残留物后加入正己烷,于12 000 r/min下高速离心10 min,取下层液体过膜上机测定。采用Phenomenex Luna C18色谱柱分离,荧光检测器检测,激发波长为241 nm,发射波长为353 nm,流动相为0.1%甲酸乙腈-0.1%甲酸水(18∶82,V/V)。结果显示,卡拉洛尔在1 ng/mL^100 ng/mL浓度范围内线性关系良好,相关系数(r)为0.999 98;添加回收率均在70%以上,相对标准偏差(RSD)均小于10%(n=5);肌肉检测限为1.25μg/kg,定量限为4.5μg/kg;肝脏检测限为2μg/kg,定量限为6.5μg/kg。

A high-performance liquid chromatography method with fluorescence detection was developed for the determination of carazolol in swine muscle and liver.Carazolol was extracted by acetonitrile,and the extract was concentrated and purified after hexane degreasing twice,the drug was finally dissolved in the acetonitrile-water（2∶8,V/V）,and the solution was centrifuged at 12 000 r/min for 10 min after adding hexane.The subnatant could be detected by high-performance liquid chromatography after being filtrated by filtration membrane.The analyte was separated by Phenomenex Luna C18 chromatographic column,detected by fluorescence detection performing at an excitation wavelength of 241 nm and an emission wavelength of 353 nm.The mobile phase was 0.1%acetonitrile/formic acid-0.1%water/formic acid（18∶82,V/V）.Results showed that the standard curve for carazolol was in good linearity in the range of 1-100 ng/mL,the correlation coefficients（r） was 0.99998.The recovery rate for carazolol wasmore than 70%,and the relative standard deviation（RSD） was less than 10%（n=5）.The limit of detection（S/N≥3） and limit of quantification（S/N≥10） by the present method for carazolol were found to be 1.25 μg/kg and 4.5 μg/kg in swine muscle,while 2 μg/kg and 6.5 μg/kg in swine liver respectively.