QuEChERS-超高效液相色谱-串联质谱法测定鹿鞭中13种类固醇激素

Simultaneous determination of 13 hormones in Testis et Penis Cervi using QuEChERS-ultra-high performance liquid chromatography-tandem mass spectrometry

该文基于QuEChERS-超高效液相色谱-串联质谱法测定了鹿鞭中13种类固醇激素(诺龙、雄烯二酮、甲睾酮、睾酮、炔诺酮、甲羟孕酮、孕酮、己烯雌酚、己烷雌酚、雌二醇、雌三醇、可的松、氢化可的松)的含量。样品经甲醇提取,QuEChERS前处理方法净化,经ACQUITY BEH C_(18)色谱柱(2.1 mm×150 mm, 3.5μm)分离,分别在电喷雾正、负离子模式下以多反应监测模式(multiple reaction monitoring, MRM)进行测定。结果显示,不同时期梅花鹿鹿鞭与马鹿鹿鞭及睾丸中激素含量存在差异。发情期的梅花鹿鹿鞭中睾酮和氢化可的松分别为10.88、12.82μg·kg^(-1),显著高于生茸期梅花鹿鹿鞭中睾酮和氢化可的松质量分数(分别为1.05、0.73μg·kg^(-1))。马鹿鹿鞭中孕酮检出量较高(6.07μg·kg^(-1)),显著高于梅花鹿鹿鞭。马鹿睾丸中孕酮最高达27.46μg·kg^(-1),是马鹿鹿鞭中的4.5倍。该检测方法前处理过程简单,灵敏度、准确度及精密度均符合要求,适用于动物源性食品中激素的检测。

A reliable QuEChERS-ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) analysis method was developed for the simultaneous determination of 13 steroid hormones(nrolone, androstenedione, methyltestosterone, testosterone, norethindrone, medroxyprogesterone, progesterone, diethylstilbestrol, hexan-stilbestrol, estradiol, estrotriol, cortisone, hydrocortisone) in Testis et Penis Cervi. The samples were extracted with methanol and purified by QuEChERS. Subsequently, the samples were separated by ACQUITY BEH C_(18)column and detected in the multiple reaction monitoring(MRM) mode under electrospray ionization in the positive and negative ion modes, respectively. Significant differences in the content of thirteen steroid hormones in Testis et Penis Cervi between the sika deer at different periods and the red deer were observed. The content of testosterone(10.88 μg·kg^(-1)) and hydrocortisone(12.82 μg·kg^(-1)) in Testis et Penis Cervi derived from rutting sika deer was significantly higher than the content of testosterone(1.05 μg·kg^(-1)) and hydrocortisone(0.73 μg·kg^(-1)) from antler growth stage. The content of progesterone in Testis et Penis Cervi derived from red deer was 6.07 μg·kg^(-1), significantly higher than that from sika deer. The content of progesterone in the testicle of red deer reached 27.46 μg·kg^(-1), 4.5 times greater than that in the penis of red deer. The sensitivity, accuracy, and precision of the method can meet the detection requirements, and the developed method is suitable for the measurement of hormones in animal-derived food.