基于UPLC-Q-Exactive Orbitrap MS结合诊断离子过滤技术快速分析石榴皮的鞣质类成分  

Rapid Analysis of Tannins from Pomegranate Peel Based on UPLC-Quadrupole/Exactive Orbitrap Mass Combined with Diagnostic Ions Filter

作　　者：刘月程 戴衍朋[1,2] 王变利 丁晓彦[1] 蒋海强[3] 周倩[1,2] LIU Yuecheng;DAI Yanpeng;WANG Bianli;DING Xiaoyan;JIANG Haiqiang;ZHOU Qian(Shandong Academy of Chinese Medicine,Jinan 250014,Shandong,China;The Key Unit for Research of Technique and Principle of Honey-processing and Carbonizing of SATCM,Jinan 250014,Shandong,China;Innovative Institute of Chinese Medicine and Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,Shandong,China)

机构地区：[1]山东省中医药研究院,山东济南250014 [2]国家中医药管理局中药蜜制和制炭技术与原理重点研究室,山东济南250014 [3]山东中医药大学中医药创新研究院,山东济南250355

出　　处：《中华中医药学刊》2023年第5期71-79,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81603299);山东省自然科学基金重点项目(ZR2020KH021);山东省重大科技创新工程项目(2020CXGC010505-04);山东省重点研发计划(软科学)项目(2021RZA07034)。

摘　　要：目的应用UPLC-Q-Exactive Orbitrap MS结合诊断离子过滤技术,对石榴皮鞣质类化学成分进行快速鉴定。方法色谱条件为Agilent ZORBAX SB-Aq C18色谱柱(3.0 mm×100 mm,1.8μm);以乙腈(A)-0.05%甲酸水(B)为流动相进行梯度洗脱(0~0.5 min,95%B;0.5~7 min,95%→75%B;7~15 min,75%B;15~30 min,75%→5%B;30~31 min,5%B;31~31.1 min,5%→95%B;31.1~35 min,95%B),流速0.3 mL·min-1,柱温30℃;质谱分析采用加热电喷雾离子源(H-ESI),正、负离子模式扫描,扫描模式为Full MS/dd-MS2,扫描范围m/z 100~1500,碰撞能20、40、60 eV。进而采用质谱数据采集-诊断离子过滤(DIF)-化合物推测及验证的研究流程对鞣质类化合物结构进行鉴定。结果石榴皮中共鉴定出61种鞣质类化合物,含鞣花酸鞣质类43种,没食子酸鞣质类10种,缩合鞣质类8种,其他鞣质类前体化合物8种。其中12种鞣质类成分首次在石榴皮中发现,包括二没食子酰葡萄糖苷及其同分异构体、三没食子酰葡萄糖苷及其同分异构体、(表)没食子儿茶素没食子酸酯、Methyl-3,4,5-trihydroxy-2-(2,3,7,8-tetrahydroxy-5,10-dioxo-5,10-dihydrochromeno[5,4,3-cde]chromen-1-yl)benzoat、(7S,8R,10R,11R,17R)-2,3,15,16,17-pentahydroxy-8-(hydroxymethyl)-5,13-dioxo-5,7,8,10,11,13-hexahydro-1,6,9,12-tetraoxa-7,11-methanocyclotrideca[def]fluoren-10-yl 3,4,5-trihydroxybenzoate、4-Rhamnopyranosyl-ellagic acid、四没食子酰葡萄糖苷及其同分异构体。结论该方法可系统、准确、快速地对石榴皮的鞣质类成分进行表征,为石榴皮药效物质研究和质量评价的进一步研究奠定基础。Objective To analyze tannins in pomegranate peel by ultra-high performance liquid chromatography-quadrupole/exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS)combined with diagnostic ion filtration technology.Methods The methanol extract of pomegranate peel was analyzed on a Agilent ZORBAX SB-Aq C18 column(3.0 mm×100 mm,1.8μm)with the mobile phase of acetonitrile(A)and 0.05%formic acid aqueous solution(B)for gradient elution(0~0.5 min,95%B;0.5~7 min,95%→75%B;7~15 min,75%B;15~30 min,75%→5%B;30~31 min,5%B;31~31.1 min,5%→95%B;31.1~35 min,95%B),the flow rate was 0.3 mL·min-1,and the column temperature was 30℃.Mass spectrometric data was obtained in positive and negative ion modes by heated electrospray ion source(H-ESI),the scanning range was m/z 100~1500 and the collision energies were 20,40,60 eV.The tannins in pomegranate peel were analyzed by the process of mass spectrometry data collection-diagnostic ion filter(DIF)-mass spectrometry identification and verification.Results A total of 61 tannins were identified,including 43 ellagic acid tannins,10 gallic acid tannins,8 condensed tannins and 8 other compounds.Among them,12 tannins were found in pomegranate peel for the first time,including bis-Galloylglucose/Isomer,tri-Galloyl-glucose/Isomer,(-)-Epicatechin gallate/(-)-Catechin gallate,Methyl-3,4,5-trihydroxy-2-(2,3,7,8-tetrahydroxy-5,10-dioxo-5,10-dihydrochromeno[5,4,3-cde]chromen-1-yl)benzoate,4-Rhamnopyranosyl-ellagic acid,(7S,8R,10R,11R,17R)-2,3,15,16,17-pentahydroxy-8-(hydroxymethyl)-5,13-dioxo-5,7,8,10,11,13-hexahydro-1,6,9,12-tetraoxa-7,11-methanocyclotrideca[def]fluoren-10-yl 3,4,5-trihydroxybenzoate and tetragalloylglucose/isomer.Conclusion The tannins in pomegranate peel can be identified accurately and rapidly by this method.This paper can provide a reference for the further research on pharmacodynamic substances and quality evaluation of pomegranate peel.

关 键 词：石榴皮 超高效液相色谱-四级杆-静电场轨道阱高分辨质谱(UPLC-Q-Exactive Orbitrap MS) 诊断离子过滤 鞣质类化合物 

分 类 号：R284.1[医药卫生—中药学]