UPLC-MS/MS法分析旋覆花蜜炙后化學成分的變化

呂渭升 位翠杰 潘曉君 楊文惠 何民友 陳向東 孫冬梅 魏梅 李振雨

中圖分類號 R284 文獻標志碼 A 文章編號 1001-0408（2021）20-2478-07

DOI 10.6039/j.issn.1001-0408.2021.20.08

摘 要 目的：比較旋覆花蜜炙前后的化學成分差異。方法：采用超高效液相色譜-串聯質譜技術（UPLC-MS/MS）。色譜柱為Waters ACQUITY UPLC BEH C18，流動相為0.1%甲酸溶液-乙腈（梯度洗脫），流速為0.3 mL/min，柱溫為30 ℃，進樣量為5 ?L。采用電噴霧離子源，以正離子模式掃描，一級質譜掃描范圍為m/z 70～1 050，二級質譜掃描范圍為m/z 50～1 050，歸一化碰撞能量為40、60 eV，質譜圖類型為峰形圖，鞘氣流速為35 arb，輔助氣流速為10 arb，噴霧電壓為3.80 kV，透鏡電壓為50 V，加熱溫度為350 ℃，毛細管溫度為350 ℃。采用Qual Browser 4.1.39.1軟件并參考在線高分辨數據庫mzCloud、中藥成分高分辨質譜本地數據庫OTCML，同時結合相關文獻對化合物進行鑒定。采用SIMCA 14.1軟件進行主成分分析（PCA）、正交偏最小二乘法-判別分析（OPLS-DA），以變量重要性投影（VIP）值大于1為標準篩選旋覆花蜜炙前后的差異性成分。結果：從旋覆花及其蜜炙品中共鑒定出29個共有化學成分，包括1-咖啡酰奎寧酸、綠原酸、3，5-二咖啡酰奎寧酸等5個酚酸類成分，槲皮素、木犀草素、狹葉依瓦菊素等12個黃酮類成分，以及1-O-乙酰旋覆花內酯、二大花旋覆花內酯B、1-O-乙?；?6-O-異丁?；蠡ㄐ不▋弱サ?2個倍半萜內酯類成分。PCA結果顯示，旋覆花與蜜旋覆花分別位于得分圖的兩側。OPLS-DA結果顯示，有7個成分的VIP值大于1，分別為峰19（大花旋覆花素）、峰6（槲皮萬壽菊苷）、峰1（1-咖啡?？鼘幩幔?、峰21（紫花牡荊素）、峰20（tomentosin）、峰13（菠葉素）、峰3（瑞香素）。結論：經蜜炙后，旋覆花的化學成分含量有所改變，且均有一定程度的降低。大花旋覆花素、槲皮萬壽菊苷、1-咖啡酰奎寧酸、tomentosin、紫花牡荊素、菠葉素、瑞香素可能為旋覆花及其蜜炙品的差異性成分。

關鍵詞 旋覆花;蜜旋覆花;超高效液相色譜-串聯質譜法;主成分分析;正交偏最小二乘法-判別分析;化學成分

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.