基于UPLC-Q-TOF MS技术快速鉴定蒸制广佛手化学成分
Rapid Analysis Compositions of Processed Citrus medica L. var. sarcodactylis Swingle by UPLC-Q-TOF MS
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摘要: 利用超高效液相色谱-四极杆-串联飞行时间质谱法（UPLC-Q-TOF MS）分析蒸制广佛手甲醇提取物的化学成分，采用Poroshell 120 EC-C18色谱柱(3.0 mm×150 mm×2.7 μm)分离，以0.1%甲酸-5 mmol/L乙酸铵水溶液乙腈为流动相进行梯度洗脱，在正、负离子模式下采集质谱数据。使用MassHunter Qualitative Analysis B.07.00软件解析各色谱峰的精确质量数、分子式、特征碎片离子，并结合相关文献及安捷伦中药化合物数据库等对蒸制广佛手成分进行分析鉴定，同时研究其质谱裂解机理。结果共鉴定出67个化学成分，包括12个黄酮及黄酮苷、22个香豆素、15个有机酸、6个糖苷类、5个柠檬苦素、7个其他成分；其中有24个化学成分为首次在佛手中检出。本研究较为全面地阐明了蒸制广佛手的化学成分，为其质量标准以及药效物质研究提供了数据支持。
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关键词:
- 蒸制广佛手 /
- 超高效液相色谱-四极杆-串联飞行时间质谱（UPLC-Q-TOF MS） /
- 化学成分鉴定 /
- 裂解机理
Abstract: Citrus medica L. var. sarcodactylis Swingle is an important and widely used herbal medicine, which has been used in China for thousands of years, and has effects of antitumor, immunomodulatory, antioxidative, anti-inflammatory, hypoglycemic, hypolipidemic and antitussive. The processed Citrus medica L. var. sarcodactylis Swingle (PCM), as Lingnan characteristic traditional Chinese sliced herbal medicine, has similar effects but less pungent, is more popular than the raw products in the Lingnan area, especially in Guangdong Provence. The technique of ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF MS) is widely and effectively used for the identification of chemical components from traditional Chinese medicine, which is considered as a forceful analysis technology in this field due to its rapid separation, accurate mass measurement and tandem mass spectrometry. In this study, UPLC-Q-TOF MS technology was used to analyze and identify the chemical components of PCM. The sample was extracted by methanol and the chromatographic separation was performed on a Poroshell 120 EC-C18 column (3.0 mm×150 mm×2.7 μm）. The mobile phase was consisted of eluent A (0.1% formic acid and 5 mmol/L ammonium acetate in water) and eluent B (acetonitrile) with the flow rate of 0.35 mL/min. The column temperature was 30 ℃, the chamber temperature was 25 ℃, and the injection volume was 6 μL at negative mode and 4 μL at positive mode, respectively. Electrospray ionization (ESI) was applied for scanning under positive and negative ion modes with the scanning range of m/z 100-1 000. The main chromatographic peaks in the total ion chromatogram (TIC) were characterized at positive and negative ion modes. Based on accurate mass, fragment ions, retention behaviors, previous reports, network database, reference standards and fragmentation pathways of compounds, a total of 67 components were identified, including 12 flavonoids and flavonoid glycosides, 20 coumarins, 15 organic acids, 6 glycosides, 5 limonin, and 7 other compounds. 24 compounds were first discovered from Citrus medica L. var. sarcodactylis Swingle, including quinic acid, malic acid, citric acid, protocatechuic acid, sucrose, catechol, esculetol, luteolin-6, 8-di-C-glucosid, 4-hydroxybenzaldehyde, eriocidin, chrysoeriol-6, 8-di-C-glucoside, diosmetin-6-C-glucoside, p-hydroxycinnamic acid, scopoletin and its isomers, azelaic acid, isofraxidin and its isomers, deacetylnomilinic acid, nomilinic acid, heraclenin, ricinoleic acid, (E, E)-9-oxooctadeca-10, 12-dienoic acid. This method is suitable for the rapid detection of compounds for PCM with easy operation, high sensitivity and excellent precision. The chemical substances of PCM were comprehensively clarified. The result provides valuable information for its quality control and pharmacological application. -
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