不同炒制温度和时间对蒺藜中4个薯蓣皂苷类成分的影响

Effects of Different Stir-Frying Temperature and Time on the Contents of Four Dioscin Components of Tribuli Fructus

目的:建立UHPLC-MS/MS法同时测定蒺藜及炒蒺藜中薯蓣皂苷元、薯蓣皂苷、伪原薯蓣皂苷和原薯蓣皂苷的含量,并研究其在炒制过程中随炒制温度(160、180、200、220℃)和时间(5、10、15、20、25 min)的变化规律。方法:采用Halo~?C(100 mm×2.1 mm,2.7μm)色谱柱;以0.1%甲酸溶液-含0.1%甲酸的乙腈为流动相,梯度洗脱;流速为0.3 mL/min;柱温为30℃;进样量为3μL;采用电喷雾离子源,多反应监测模式,正离子扫描。结果:所测4个成分在相应的浓度范围内与峰面积的线性关系良好(r≥0.9998),平均加样回收率为97.08%~98.15%,RSD为2.57%~3.86%。炒制前后4个成分含量有明显差异,随炒制温度升高和炒制时间延长,薯蓣皂苷元含量从无到有且逐渐升高,薯蓣皂苷和原薯蓣皂苷含量逐渐下降。160℃时,随炒制时间延长,伪原薯蓣皂苷含量持续上升,180、200、220℃时,其含量先升高后降低。结论:该研究建立的UHPLC-MS/MS法适用于薯蓣皂苷元、薯蓣皂苷、伪原薯蓣皂苷和原薯蓣皂苷的同时测定,梳理出这4个成分在蒺藜炒制过程中的变化规律,可为进一步揭示蒺藜的炮制原理提供参考。

Objective:To establish the UHPLC-MS/MS method for simultaneous determination of diosgenin,dioscin,pseudoprotodioscin and protodioscin in Tribuli Fructus and its fried products,and to study the changes of the compounds in Tribuli Fructus with stir-frying temperature(160,180,200,220℃)and time(5,10,15,20,25 min).Methods:A HaloC(100 mm×2.1 mm,2.7μm)column was used.Gradient elution was performed with 0.1%formic acid solution and acetonitrile containing 0.1%formic acid as mobile phase.The flow rate was 0.3 mL/min.The column temperature was 30℃.The injection volume was 3μL.Multiple-reaction monitoring(MRM)was employed with switching electron spray ionization source in positive mode.Results:The linear relationship between the peak area and the concentration range of the four components was good(r≥0.9998).The average recoveries ranged from 97.08%to 98.15%with RSD of 2.57%to 3.86%.The contents of 4 components were obviously different before and after frying.The contents of diosgenin increased gradually with the increase of cooking temperature and cooking time,while the contents of dioscin and protodioscin decreased gradually.At 160℃,the content of pseudoprotodioscin continued to increase with the extension of frying time,at 180,200 and 220℃,its content first increased and then decreased.Conclusion:The UHPLC-MS/MS method established in this study is suitable for simultaneous determination of diosgenin,dioscin,pseudoprotodioscin and protodioscin.The changes of these four components in the frying process of Tribuli Fructus are analyzed,which can provide a reference for further revealing the processing principle of Tribuli Fructus.