鸡肌肉组织中醋酸甲羟孕酮高效液相色谱检测方法的建立

Development of high performance liquid chromatography for detection of medroxyprogesterone acetate in the chicken muscle

本研究建立了一种鸡肌肉组织中醋酸甲羟孕酮残留的高效液相色谱检测方法。样品经乙腈提取,正己烷去除蛋白和脂类物质,通过中性氧化铝小柱净化,采用乙腈洗脱后氮气吹干,流动相复溶后过滤上高效液相色谱仪测定。醋酸甲羟孕酮在20~500 ng/mL的浓度范围内呈线性相关,相关系数为0.999。该方法对醋酸甲羟孕酮在空白鸡肌肉组织中的定量限为0.01μg/kg;醋酸甲羟孕酮在0.01、0.02、0.04μg/kg的浓度添加水平上,其平均回收率在70.0%~90.5%,批间、批内相对标准偏差均小于10%。此方法准确、可靠,适用于鸡肌肉组织中醋酸甲羟孕酮的残留检测。

In this study,a high performance liquid chromatography（HPLC） method was established to detect medroxyprogesterone acetate residues in chicken muscle.The sample were extracted by ethyl acetate,defatted with n-hexane,purified with neutral alumina solid-phase extraction cartridges,eluted with acetonitrile and then dried with nitrogen.The residue was dissolved with the mobile phase and then filtered.It was then analyzed with HPLC.A good linearity of the calibration curves for medroxyprogesterone acetate was obtained within the ranges of 20 to 500 ng/mL（R2= 0.9996）.The limit detection of the method was 0.01μg/kg.The average recoveries of medroxyprogesterone acetate were 70.0 to 90.5%at three spiked levels（0.01,0.02 and 0.04 μg/kg） with the intra and inter RSD lower than 10%.The method is accurate and reliable,and could be applied to detect medroxyprogesterone acetate residue in the chicken muscle.