UPLC-MS/MS法分析旋覆花蜜炙后化学成分的变化

吕渭升 位翠杰 潘晓君 杨文惠 何民友 陈向东 孙冬梅 魏梅 李振雨

中圖分类号 R284 文献标志码 A 文章编号 1001-0408（2021）20-2478-07

DOI 10.6039/j.issn.1001-0408.2021.20.08

摘 要 目的：比较旋覆花蜜炙前后的化学成分差异。方法：采用超高效液相色谱-串联质谱技术（UPLC-MS/MS）。色谱柱为Waters ACQUITY UPLC BEH C18，流动相为0.1%甲酸溶液-乙腈（梯度洗脱），流速为0.3 mL/min，柱温为30 ℃，进样量为5 ?L。采用电喷雾离子源，以正离子模式扫描，一级质谱扫描范围为m/z 70～1 050，二级质谱扫描范围为m/z 50～1 050，归一化碰撞能量为40、60 eV，质谱图类型为峰形图，鞘气流速为35 arb，辅助气流速为10 arb，喷雾电压为3.80 kV，透镜电压为50 V，加热温度为350 ℃，毛细管温度为350 ℃。采用Qual Browser 4.1.39.1软件并参考在线高分辨数据库mzCloud、中药成分高分辨质谱本地数据库OTCML，同时结合相关文献对化合物进行鉴定。采用SIMCA 14.1软件进行主成分分析（PCA）、正交偏最小二乘法-判别分析（OPLS-DA），以变量重要性投影（VIP）值大于1为标准筛选旋覆花蜜炙前后的差异性成分。结果：从旋覆花及其蜜炙品中共鉴定出29个共有化学成分，包括1-咖啡酰奎宁酸、绿原酸、3，5-二咖啡酰奎宁酸等5个酚酸类成分，槲皮素、木犀草素、狭叶依瓦菊素等12个黄酮类成分，以及1-O-乙酰旋覆花内酯、二大花旋覆花内酯B、1-O-乙酰基-6-O-异丁酰基大花旋覆花内酯等12个倍半萜内酯类成分。PCA结果显示，旋覆花与蜜旋覆花分别位于得分图的两侧。OPLS-DA结果显示，有7个成分的VIP值大于1，分别为峰19（大花旋覆花素）、峰6（槲皮万寿菊苷）、峰1（1-咖啡酰奎宁酸）、峰21（紫花牡荆素）、峰20（tomentosin）、峰13（菠叶素）、峰3（瑞香素）。结论：经蜜炙后，旋覆花的化学成分含量有所改变，且均有一定程度的降低。大花旋覆花素、槲皮万寿菊苷、1-咖啡酰奎宁酸、tomentosin、紫花牡荆素、菠叶素、瑞香素可能为旋覆花及其蜜炙品的差异性成分。

关键词 旋覆花;蜜旋覆花;超高效液相色谱-串联质谱法;主成分分析;正交偏最小二乘法-判别分析;化学成分

Variations of Chemical Components in Inula japonica by UPLC-MS/MS before and after Honey-frying

LYU Weisheng1，2，WEI Cuijie1，2，PAN Xiaojun1，2，YANG Wenhui1，2，HE Minyou1，2，CHEN Xiangdong1，2，SUN Dongmei1，2，WEI Mei1，2，LI Zhenyu1，2（1. Guangdong e-fong Pharmaceutical Co.， Ltd.， Guangdong Foshan 528244， China; 2. Guangdong Provincial Key Laboratory of TCM Formula， Guangdong Foshan 528244， China）

ABSTRACT   OBJECTIVE： To compare the chemical components differences of Inula japonica before and after honey-frying. METHODS： UPLC-MS/MS method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C18 column with mobile phase consisted of 0.1% formic acid-acetonitrile （gradient elution） at the flow rate of 0.3 mL/min. The column temperature was set at 30 ℃， and sample size was 5 ?L. The electrospray ion source was scanned by positive ion mode. The first order mass spectrometry scanning range was m/z 70-1 050， the second order mass spectrometry scanning range was m/z 50-1 050， and the normalized collision energy was 40， 60 eV ; mass spectrum type was the peak figure， the flow rate of sheath gas was 35 arb， the auxiliary airflow speed was 10 arb， the spray voltage was 3.80 kV， the S-lens voltage was 50 V， the heating temperature was 350 ℃， and the capillary temperature was 350 ℃. The components were identified by Qual Browser 4.1.39.1 software， referring to the online high-resolution database mzCloud and local database OTCML of high-resolution mass spectrometry of TCM， and combined with relevant literature. The principal component analysis （PCA） and orthogonal partial least squared-discriminant analysis （OPLS-DA） of I. japonica before and after honey-fried were performed by using SIMCA 14.1 statistical software， and variable importance projection （VIP） value greater than 1 was used as the standard to screen the differential components before and after honey-frying. RESULTS： A total of 29 common chemical components were identified from I. japonica and honey-fried I. japonica， including 5 phenolic acids as 1-caffeoylquinic acid， chlorogenic acid and 3，5-dicaffeoylquinic acid， 12 flavonoids as quercetin， luteolin and evamectin， as well as 12 sesquiterpene lactones as 1-O-acetylinula diester， inula bicolor lactone B and 1-O-acetyl-6-O-isobutyryl inulin. The results of PCA showed that I. japonica and honey-fried I. japonica were located on both sides of the score diagram respectively. The results of OPLS-DA showed that the VIP values of 7 components were greater than 1， which were peak 19 （britanin）， peak 6 （quercetagitrin）， peak 1 （1-caffeoylquinic acid）， peak 21 （vitexicarpin）， peak 20 （tomentosin）， peak 13 （spinacetin） and peak 3 （daphnetin）. CONCLUSIONS： After honey-fried， the content of chemical components of I. japonica changed and decreased to a certain extent. Britanin， quercetagitrin， 1-caffeoylquinic acid， tomentosin，vitexicarpin， spinacetin and daphnetin may be the differential components of I. japonica and honey-fried I. japonica.