目的 建立一种用于快速准确检测抗疲劳类保健食品中非法添加9种壮阳类化学药物(甲磺酸酚妥拉明、甲睾酮、司坦唑醇、达那唑、他达拉非、枸橼酸西地那非、爱地那非、伐地那非、硫代艾地那非)的方法。方法 采用UPLC-MS/MS法, 以Waters Acquity BEH-C₁₈柱(100 mm×2.1 mm, 1.7 μm)为色谱柱, 以0.1%甲酸甲醇溶液(A)-0.1%甲酸水溶液(B)为流动相, 梯度洗脱:0～5 min, 50% A;5～7 min, 50%～90% A;7～9 min, 90%～100% A;9～10 min, 100%～50% A;体积流量0.2 mL/min, 进样量5 μL, 柱温40 ℃。选择ESI离子源、正离子扫描、多反应监测(MRM)模式测定9种临床常用的化学药物, 通过比较MRM通道中样品峰与对照品峰的分子离子峰、二级碎片离子峰、色谱保留时间等信息确定添加的化学药物, 并根据外标法以质谱峰面积计算添加药物的准确量。结果 在相应色谱及质谱条件下, 9种化学药物甲磺酸酚妥拉明、甲睾酮、司坦唑醇、达那唑、他达拉非、枸橼酸西地那非、爱地那非、伐地那非、硫代艾地那非的分离度良好, 方法检测限(LOD)均在0.1～0.3 ng/g, 定量限(LOQ)均在0.3～0.9 ng/g, 标准加样回收率均在88.4%～116.3%。68批抗疲劳类保健食品中检出41批添加了化学药, 阳性率为60.3%;样品中检出了枸橼酸西地那非、他达拉非、爱地那非、硫代艾地那非。结论 本法简便、准确, 灵敏度高, 可作为抗疲劳类保健食品中非法添加化学药物的定性定量检测方法。

Objective To establish an accurate method for the determination of nine chemical drugs (phentolamine mesylate, methyltestosterone, stanozolol, danazol, tadalafil, sildenafil citrate, aildenafil, vardenafil, and thioaildenafil) which were illegally added into the antifatigue health foods. Methods The UPLC-MS/MS method was adopted. The samples were extracted with methanol by ultrasonic processing and separated on a Waters Acquity BEH-C₁₈ (100 mm × 2.1 mm, 1.7 μm) column with 0.1% formic acid methanol (A) and 0.1% formic acid water (B) as the mobile phase by gradient elution (0—5 min, 50% A; 5—7 min, 50%—90% A; 7—9 min, 90%—100% A; 9—10 min, 100%—50% A) at a flow rate of 0.2 mL/min. The injection volume was 5 μL. The column temperature was 40 ℃. The positive-ion (ESI⁺) source and MRM mode were used to separate and quantitatively determine the chemicals. The obtained molecular ions, fragment ions, and retention time for MRM channels were used to identify the nine kinds of drugs by comparing with those of reference substances. The obtained peak areas were used to determine the accurate content of the nine chemicals in the antifatigue health foods. Results A good resolution of the nine kinds of chemical drugs, including phentolamine mesylate, methyltestosterone, stanozolol, danazol, tadalafil, sildenafil citrate, aildenafil, vardenafil, and thioaildenafil, was obtained under this UPLC and MS/MS conditions. The limits of detection (LOD) and quantification (LOQ) were in the ranges of 0.1—0.3 ng/g and 0.3—0.9 ng/g. The standard addition recoveries were in the range of 88.4%—116.3%. There were 68 batches of antifatigue health foods, among which 41 batches were added with the chemicals with positive rate of 60.3%. The sildenafil citrate, tadalafil, aildenafil, and hioaildenafil were detected in samples. Conclusion The method is simple, accurate, and has high sensitivity, which can be used for the qualitative and quantitative determination of illegally added chemical drugs in the antifatigue health foods.

浙江省公益性技术应用研究计划资助项目(2013C37103);"十二五"浙江省高校重点学科(药理学)资助项目