HPLC-MS/MS法同时测定育发液中的9种禁限用物质

Simultaneous determination of 9 restricted substances in hair growth liquid by HPLC-MS/MS

目的:建立可同时检测育发液中斑蝥素、米诺地尔、雌三醇、雌二醇、睾酮、甲睾酮、雌酮、己烯雌酚、黄体酮9种禁限用物质的液相色谱-质谱法。方法:采用CAPCELL PAK MG C_(18)色谱柱(150 mm×2 mm,5μm)进行分离,以10 mmol·L^(-1)醋酸铵溶液(A)-乙腈(B)为流动相进行梯度洗脱(0~12 min,90%A→50%A;12~13 min,50%A→60%A;13~20 min,60%A;20~30 min,60%A→20%A;30~31 min,20%A→90%A;31~40 min,90%A),流速0.2 m L·min^(-1);使用加热电喷雾离子源,正负离子检测,采用选择反应监测扫描方式。结果:斑蝥素、米诺地尔、雌三醇、雌二醇、睾酮、甲睾酮、雌酮、己烯雌酚、黄体酮质量浓度分别在1~100、0.001~0.05、0.25~25、15~300、0.05~5、0.1~5、0.06~3、0.02~2、0.05~5μg·m L^(-1)范围内均有良好的线性关系(r≥0.998);最低检出浓度分别为1.25、0.001、0.25、15、0.05、0.1、0.08、0.02、0.05μg·g^(-1),最低定量浓度分别为5、0.005、1.25、75、0.25、0.5、0.3、0.1、0.25μg·g^(-1)。在12批育发液样品中检出2批非法添加了米诺地尔。结论:本研究建立的方法可同时定性定量检测育发液中性激素、斑蝥素与米诺地尔等9种禁限用物质。

Objective：To establish a method for the simultaneous determination of 9 restricted substances（cantharidin,minoxidil,estriol,estradiol,testosterone,methyltestosterone,estrone,diethylstilbestrol and progesterone）in hair growth liquid by using high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS）.Methods：The HPLC separation was performed on a CAPCELL PAK MG C_（18） column（150 mm×2 mm,5 μm）.The mobile phase was 10 mmol·L^（-1） ammonium acetate solution（A）-acetonitrile（B）for gradient elution（0^（-1）2 min,90%A → 50%A;12^（-1）3 min,50%A → 60%A;13-20 min,60%A;20-30 min,60%A → 20%A;30-31 min,20%A → 90%A;31-40 min,90%A）,and the flow rate was 0.2 m L·min^（-1）.Ion source was heat assisted electrospray ion source（HESI）.Positive ion and negative ion were used for detection with a scanning mode of selective reaction monitoring（SRM）.Results：A good linear correlation was observed over the range of 1^（-1）00 μg·m L^（-1） for cantharidin,0.001-0.05 μg·m L^（-1） for minoxidil,0.25-25 μg·m L^（-1） for estriol,15-300 μg·m L^（-1） for estradiol,0.05-5 μg·m L^（-1） for testosterone,0.1-5 μg·m L^（-1） for methyltestosterone,0.06-3 μg·m L^（-1） for estrone,0.02-2 μg·m L^（-1） for diethylstilbestrol,0.05-5 μg·m L^（-1） for progesterone（r ≥0.998）.The limits of detection of 9 restricted substances were 1.25 μg·g^（-1）,0.001 μg·g^（-1）,0.25 μg·g^（-1）,15 μg·g^（-1）,0.05 μg·g^（-1）,0.1 μg·g^（-1）,0.08 μg·g^（-1）,0.02 μg·g^（-1） and 0.05 μg·g^（-1）,respectively.The limit of quantification of 9 restricted substances were 5 μg·g^（-1）,0.005 μg·g^（-1）,1.25 μg·g^（-1）,75 μg·g^（-1）,0.25 μg·g^（-1）,0.5 μg·g^（-1）,0.3 μg·g^（-1）,0.1 μg·g^（-1） and 0.25 μg·g^（-1）,respectively.Totally,2 of 12 tested hair growth liquid samples contained illegally added minoxidil.Conclusion：The method can be used for simultaneous qualitative and quantitative determination of 9 restricted substances（sex hormones,cantharidin and minoxidil）in hair growth liquid.