分散微固相萃取-超高效液相色谱-高分辨质谱法测定火锅底料中罂粟壳生物碱

Determination of poppy shell alkaloids in hot pot condiment by ultra-high performance liquid chromatography-high resolution mass spectrometry coupled with dispersive micro solid-phase extraction

目的建立火锅底料中5种罂粟壳生物碱的超高效液相色谱-高分辨质谱检测方法。方法样品经基于强阳离子交换吸附剂(PCX)的分散微固相萃取(DMSPE)提取和净化后,以0.1%甲酸水溶液-0.1%甲酸乙腈为流动相进行梯度洗脱,HSS T3色谱柱(2.1 mm×100 mm, 1.8μm)分离,高分辨质谱平行反应监测模式采集,外标法定量。结果在给定的浓度范围内,5种罂粟壳生物碱呈良好的线性关系(R^(2)>0.999);方法检出限为0.1~0.3μg/kg,定量限为0.4~1.0μg/kg。火锅底料在1、10、100μg/kg不同加标水平中的回收率在78.5%~104.2%之间,日内精密度(RSDr)在1.8%~6.8%之间,日间精密度(RSD_(R))在4.9%~13.7%之间。结论本方法采用以PCX为吸附剂的DMSPE净化技术,在保证有效净化的基础上,简单快速、成本较低,方法灵敏度高、重现性好,可应用于固体和半固体火锅底料中5种罂粟壳生物碱的定性定量检测。

Objective To develop an analytical method for determination of 5 kinds of poppy shell alkaloids in hot pot condiment by ultra-high performance liquid chromatography-high resolution mass spectrometry. Methods The samples were extracted and cleaned up based on dispersive micro solid-phase extraction(DMSPE) using strong cationic exchange adsorbent(PCX). The chromatographic separation of alkaloids was performed on an HSS T3 column(2.1 mm×100 mm, 1.8 μm) using 0.1% formic acid aqueous solution and acetonitrile with 0.1% formic acid as the mobile phase with gradient elution. The alkaloids were analyzed by high resolution mass spectrometry using parallel reaction monitoring mode and the external standard method was selected for quantitative analysis. Results Five kinds of alkaloids showed a good linearity in a certain concentration range with relative coefficient of R^(2)>0.999. Limits of detections(LODs) were in the range of 0.1-0.3 μg/kg, and limits of quantitations(LOQs) were in the range of 0.4-1.0 μg/kg. At the spiked levels of 1, 10, 100 μg/kg, the recoveries of alkaloids in hot pot condiment were from 78.5% to 104.2%, with the intra-day precision(RSDr) of 1.8%-6.8% and the inter-day precision(RSDR) of 4.9%-13.7%. Conclusion Using DMSPE cleanup technique with PCX as adsorbent, this method was simple, rapid, low-cost, high sensitivity, and had good reproducibility on the basis of ensuring effective cleanup effect. It could be applied to the qualitative and quantitative determination of five poppy shell alkaloids in hot pot condiment.