液相色谱-质谱联用法(HPLC-MS/MS)快速测定凉茶中的5种罂粟壳类生物碱

Rapid Determination of Five Alkaloids in Poppy Shell in Herbal Tea by HPLC-MS/MS

目的:建立液相色谱-质谱联用法(HPLC-MS/MS)快速检测凉茶中非法添加的吗啡、可待因、蒂巴因、罂粟碱、那可丁等5种罂粟壳类生物碱。方法:采用Agilent Poroshell 120 EC-C;(2.1 mm×100 mm,2.7μm)色谱柱分离,含0.1%乙酸的5 mmol/L乙酸铵溶液-甲醇为流动相梯度洗脱,在电喷雾离子源正离子模式下,采用多反应监测(MRM)方式测定,外标法定量。结果:吗啡、可待因的线性范围为2.5~100 ng/mL,蒂巴因、罂粟碱、那可丁的线性范围为0.5~20 ng/mL,5种罂粟壳类生物碱的线性相关系数均大于0.995(R>0.995)。检出限吗啡、可待因为0.8μg/g,罂粟碱、蒂巴因、那可丁为0.2μg/g。5种生物碱成分的3个添加水平在液体基质中回收率为84.5%~105.1%,RSD不大于6.3%(n=6),固体基质中回收率为72.5%~112.3%,RSD不大于7.1%(n=6)。应用方法对150批凉茶样品进行了抽检,其中6批检出。结论:该方法操作简便,准确可靠,适用于快速分析测定凉茶中的5种罂粟壳类成分。

Objective: A HPLC-MS/MS method was developed for rapid determination of five alkaloids in Poppy Shell in herbal tea, including morphine, codeine, thebaine, papaverine, and narcotine. Method: Agilent Poroshell 120 EC-C;(2.1 mm×100 mm,2.7 μm)was applied with the mobile phase consisting of 5 mmol/L ammonium acetate containing 0.1 %acetic acid-methanol by gradient elution. The electrospray ion source was operated in the positive ion mode using the multireaction monitoring(MRM) method, and the results were quantified by the external standard method. Result: The concentration range was 2.5~100 ng/mL in morphine and codeine, 0.5~20 ng/mL in the others. The correlation coefficients(R) were all above 0.995. The detection limit was 0.8 μg/g in morphine and codeine, 0.2 μg/g in the others. The average recoveries ranged from 84.5% to 105.1% in liquid samples, with relative stand deviations(RSDs) all below 6.3%(n=6),ranged from 72.5% to 112.3% in solid samples, with relative stand deviations(RSDs) all below 7.1%(n=6). 150 Batches of samples bought from markets were screened, and 6 batches showed positive for these analytes. Conclusion: This method was simple, accurate, could be used for the determination of five alkaloids in Poppy Shell added in herbal tea.