UPLC/Q-TOF-MS 和 PLS-DA 在鸦片生物碱检测及分类判别中的应用

目的建立了鸦片中吗啡、可待因、蒂巴因、罂粟碱、那可汀5种常量生物碱和牛心果碱、劳丹宁、劳丹素等12种痕量生物碱同时分离鉴定的超高效液相色谱-高分辨四极杆飞行时间串联质谱（UPLC/Q-TOF）方法。方法采用Agilent Eclipse Plus C18RRHD（2.1 mm×100 mm,1.8μm）色谱柱,以10 mM甲酸铵溶液和乙腈体系梯度洗脱,使用ESI离子源,正离子模式下采集数据。结果借助UPLC的快速分离和Q-TOF-MS测定的精确分子量和二级碎片信息,实现了17种生物碱的检测。应用SIMCA-P软件对78份国产鸦片和200份缅甸产鸦片进行偏最小二乘法判别分析（partial least squaresdiscriminate analysis,PLS-DA）,比较不同地区鸦片样品中生物碱的差异,建立了区分国产鸦片和缅甸产鸦片的判别方法。结论所建立的方法用于鸦片样品中常量和痕量生物碱的检测及鸦片样品的产地判别,具有快速、高效、灵敏、准确的特点。

Determination of Opium Alkaloids and Discrimination of Sample Origins by UPLC/Q-TOF and PLS-DA

Objective An ultra-performance liquid chromatography coupled to quadrupole-time of flight mass spectrometry （UPLC/Q-TOF） method was developed for the simultaneous determination of 5 major opium alkaloids, i.e. morphine, codeine, thebaine, papaverine, noscapine, and 12 trace alkaloids, such as reticuline, laudanidine, laudanosine, etc. Method The chromatographic separation was performed on an Agilent Eclipse Plus C18 RRHD （2.1 mm×100 mm, 1.8 μm） column with a gradient elution of acetonitrile-lO mM ammonium formate. The mass spectrometer equipped with electrospray ionization source was used as detector in the positive ion mode. Results A method for the rapid separation and determination of 17 alkaloids by UPLC/Q-TOF was established. The differences of alkaloids between opium samples from different origins were found and a discrimination model was built by partial least squares-discriminate analysis （PLS-DA） with SIMCA-P software based on the data set of these 17 alkaloids variables in 78 domestic and 200 Myanmar opium samples. Conclusion The established method is of rapidness, high accuracy, good selectivity and sensitivity. It is suitable for the simultaneous determination of alkaloids and the discrimination of sample origins.