高分辨液质联用技术评价半夏及其炮制品硫熏前后的品质

Quality Evaluation of Pinelliae Rhizoma and Its Processed Products Before and After Sulfur Fumigation Based on UPLC-Q-TOF-MS

目的建立评价半夏及其炮制品硫熏前后品质的超高效液相色谱-四极杆-飞行时间高分辨质谱(UPLC-Q-TOF-MS)法。方法色谱柱为Waters HSS T3柱(100 mm×2.1 mm,1.7µm),流动相为甲醇-乙腈(1∶1,V/V)-0.1%甲酸溶液(梯度洗脱),在正、负离子监测模式下采集半夏及其炮制品硫熏前后的化合物数据,与在Chemspider,mzCloud,Metaboanalyst数据库及相关文献中检索到的化合物进行比对。应用Sciex OS软件对采集到的硫熏样品数据进行分析,找到含硫化合物的特征性中性丢失或特征性碎片离子,判定半夏及其炮制品硫熏后的含硫产物。结合偏最小二乘判别分析(PLS-DA)法对半夏及其炮制品进行分类。结果半夏及其炮制品中共检出254种化合物,主要为氨基酸、核苷及其衍生物、有机酸等小分子化合物,以及皂苷、生物碱、木质素衍生物等天然产物。半夏炮制品(姜半夏、法半夏、清半夏)硫熏后均可检出31种含硫化合物,其中6-姜辣素仅在姜半夏中检出,大豆苷和甘草酸仅在法半夏中检出。在不同程度硫熏半夏中,含硫量变化趋势基本符合严重硫熏、超限度>稍熏、略超限度>未熏的变化趋势。结论建立的UPLC-Q-TOF-MS法可快速、全面地分析对半夏及其炮制品中的化学成分。硫熏会影响半夏及其炮制品的品质,在半夏加工过程中应合理、规范使用硫熏技术,避免过度硫熏对半夏药材质量的影响。

Objective To establish an ultra-performance liquid chromatography-quadrupole/time-of-flight high-resolution mass spectrometry(UPLC-Q-TOF-MS)method for evaluating the quality of Pinelliae Rhizoma and its processed products before and after sulfur fumigation.Methods The chromatographic column was Waters HSS T3 column(100 mm×2.1 mm,1.7µm),the mobile phase was methanol acetonitrile(1∶1,V/V)-0.01 formic acid solution(gradient elution).Compound data of Pinelliae Rhizoma and its processed products before and after sulfur fumigation were collected under positive and negative ion monitoring modes,and those were compared with compounds retrieved from Chemspider,mzCloud,Metaboanalyst databases,and relevant literature.The Sciex OS software was used to analyze the collected data of the sample processed by sulfur fumigation,identify characteristic neutral loss or characteristic fragment ions of sulfur-containing compounds,and determine the sulfur-containing products of Pinelliae Rhizoma and its processed products after sulfur fumigation.The Pinelliae Rhizoma and its processed products were classified by the partial least squares-discriminant analysis(PLS-DA)method.Results A total of 254 compounds were identified from Pinelliae Rhizoma and its processed products,and the main compounds identified were small molecule compounds such as amino acids,nucleosides and their derivatives,organic acids,and natural products such as saponins,alkaloids and lignin derivatives.A total of 31 sulfur-containing compounds could be detected in all the processed products of Pinelliae Rhizoma(Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine,Pinelliae Rhizoma Praeparatum,and Pinelliae Rhizoma Praeparatum cum Alumine)after sulfur fumigation,among which 6-gingerol was found only in Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine,Daidzein and glycyrrhizin were found only in Pinelliae Rhizoma Praeparatum.In Pinelliae Rhizoma processed with different degrees of sulfur fumigation,the trend of peak area changes in sulfur content was basically in line with the trend of severe sulfur fumigation,exceeding the limit>slightly fumigation,and slightly exceeding the limit>no fumigation.Conclusion The established UPLC-Q-TOF-MS method can rapidly and comprehensively analyze the chemical compounds of Pinelliae Rhizoma and its processed products.Sulfur fumigation can affect the quality of Pinelliae Rhizoma and its processed products.During the processing of Pinelliae Rhizoma,sulfur fumigation technology should be used in a reasonable and standardized manner to avoid the impact of excessive sulfur fumigation on the quality of Pinelliae Rhizoma.