HPLC-MS/MS法同时测定育发液中的9种禁限用物质

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 C₁₈色谱柱（150 mm×2 mm，5 μm）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 mL·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、0.05~5 μg·mL^-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、0.05 μg·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、0.25 μg·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 C₁₈ column (150 mm×2 mm,5 μm).The mobile phase was 10 mmol·L^-1 ammonium acetate solution(A)-acetonitrile(B)for gradient elution (0-12 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),and the flow rate was 0.2 mL·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 1-100 μg·mL^-1 for cantharidin,0.001-0.05 μg·mL^-1 for minoxidil,0.25-25 μg·mL^-1 for estriol,15-300 μg·mL^-1 for estradiol,0.05-5 μg·mL^-1 for testosterone,0.1-5 μg·mL^-1 for methyltestosterone,0.06-3 μg·mL^-1 for estrone,0.02-2 μg·mL^-1 for diethylstilbestrol,0.05-5 μg·mL^-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.