基于HPLC-MS/MS的小粒咖啡花酚类活性成分分析

李倩; 刘兴勇; 米艳华; 尹本林; 陈泽历; 王雪娇; 罗刚; 宋易霖; 刘振环

DOI:10.12161/j.issn.1005-6521.2025.24.020

食品研究与开发:2025,46(24):156-164

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基于HPLC-MS/MS的小粒咖啡花酚类活性成分分析

李倩1，刘兴勇1，米艳华1，尹本林1，陈泽历1，王雪娇2，罗刚3，宋易霖4，刘振环1*

(1.云南省农业科学院质量标准与检测技术研究所,云南昆明 650205；2.云南省农业科学院农业经济信息研究所,云南昆明 650205；3.宁洱县植保植检站,云南普洱 665199；4.保山市农业机械化技术推广站,云南保山 678000)

Analysis of Active Phenols in Coffea Arabica Flowers Based on HPLC-MS/MS

LI Qian1,LIU Xingyong1,MI Yanhua1,YIN Benlin1,CHEN Zeli1,WANG Xuejiao2,LUO Gang3,SONG Yilin4,LIU Zhenhuan1*

(1.Quality Standard and Testing Technique Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；2.Agricultural Economics and Informations Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；3.Ning´er Plant Protection and Inspection Station,Pu´er 665199,Yunnan,China；4.Station of Baoshan Agricultural and Mechanizitional Extension,Baoshan 678000,Yunnan,China)

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投稿时间：2024-06-11

中文摘要: 为促进咖啡副产物的开发利用，建立高效液相色谱-串联质谱同时测定咖啡花中13种活性化合物的方法，明确其活性成分含量及抗氧化活性。使用Kinetex C18色谱柱（100 mm×4.6 mm，2.6 μm），甲醇（A）-0.3%甲酸水溶液（B）为流动相，梯度洗脱，柱温30 ℃，流速1.0 mL/min，电喷雾离子源，多反应监测模式正、负离子同时扫描。对提取溶剂、提取时间、扫描模式进行优化，并进行含量分析、相关性分析、抗氧化活性评价。结果表明，最优的提取溶剂为40%乙醇溶液，最佳提取时间为8 min，13种化合物在45 min内完成分析，化合物相关系数为0.997 3～0.999 8，回收率范围为92.5%～104.6%。活性成分含量由大到小分别为葫芦巴碱、咖啡因、绿原酸、苹果酸、异绿原酸A、柠檬酸、琥珀酸、异绿原酸C、新绿原酸、隐绿原酸、异绿原酸B、芦丁、咖啡酸。活性化合物之间具有显著相关性，除琥珀酸与咖啡因呈显著负相关外，其他活性成分之间为正相关，绿原酸与新绿原酸、异绿原酸A相关性系数分别为0.93、0.92；异绿原酸B与异绿原酸C相关性系数为0.92。绿原酸、新绿原酸、异绿原酸A、咖啡酸、异绿原酸B、芦丁为与抗氧化活性相关性系数排名前6的化合物，相关性系数分别为0.81、0.80、0.68、0.65、0.57、0.55。该研究为挖掘咖啡花活性成分及其抗氧化活性提供参考。

中文关键词: 高效液相色谱-串联质谱咖啡花酚类活性化合物抗氧化活性

Abstract:To promote the development and utilization of coffee by-products,a method was designed to detect 13 active compounds in coffee flowers by high-performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS）,determining the content of active ingredients and antioxidant activity of coffee flowers.The Kinetex C18 chromatographic column（100 mm × 4.6 mm,2.6 μm） was used.With methanol （A）-0.3% formic acid aqueous solution （B） as mobile phase,gradient elution was performed.The flow rate was 1.0 mL/min,and the column temperature was set at 30 ℃.An electrospray ionization（ESI） and simultaneous scanning of positive and negative ions in multi-reaction monitoring （MRM） mode were conducted.The extraction solvent,extraction time,and scanning mode were optimized,and quantitative analysis,correlation analysis,and antioxidant activity evaluation were carried out.The optimal extraction solvent and time were confirmed as 40% ethanol solution and 8 min.The analysis of 13 compounds was completed in 45 minutes.The correlation coefficients of compounds were between 0.997 3 and 0.999 8,and the recovery rates ranged from 92.5% to 104.6%.The content of active compounds from large to small was trigonelline,caffeine,chlorogenic acid,malic acid,isochlorogenic acid A,citric acid,succinic acid,isochlorogenic acid C,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid B,ludin,and caffeic acid.There were significant correlations among the active compounds.Except for the significant negative correlation between succinic acid and caffeine,the other active compounds were positively correlated.The correlation coefficients of chlorogenic acid with neochlorogenic acid and with isochlorogenic acid A were 0.93 and 0.92,respectively.The correlation coefficient between isochlorogenic acid B and isochlorogenic acid C was 0.92.Chlorogenic acid,neochlorogenic acid,isochlorogenic acid A,caffeic acid,isochlorogenic acid B,and rutin were identified as the top six compounds most correlated with antioxidant activity,with correlation coefficients of 0.81,0.80,0.68,0.65,0.57,and 0.55,respectively.This research offered a reference for understanding the active components and antioxidant activities of coffee flowers.

keywords: high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） coffee flower phenol active compound antioxidant activity

文章编号：202524020 中图分类号： 文献标志码：

基金项目:云南省基础研究专项（202401AT070048）；云南省农业科学院质量标准与检测技术研究所所长基金（ZBS202302）；云南省农业科学院科研预研专项（2023KYZX-17）；云南省重大科技专项（农业）计划项目（202102AE090051）；云南省科技厅创新引导与科技型企业培育计划（202304AL030013）

作者	单位
李倩	1.云南省农业科学院质量标准与检测技术研究所,云南昆明 650205；
刘兴勇	1.云南省农业科学院质量标准与检测技术研究所,云南昆明 650205；
米艳华	1.云南省农业科学院质量标准与检测技术研究所,云南昆明 650205；
尹本林	1.云南省农业科学院质量标准与检测技术研究所,云南昆明 650205；
陈泽历	1.云南省农业科学院质量标准与检测技术研究所,云南昆明 650205；
王雪娇	2.云南省农业科学院农业经济信息研究所,云南昆明 650205；
罗刚	3.宁洱县植保植检站,云南普洱 665199；
宋易霖	4.保山市农业机械化技术推广站,云南保山 678000
刘振环	1.云南省农业科学院质量标准与检测技术研究所,云南昆明 650205；

Author Name	Affiliation
LI Qian	1.Quality Standard and Testing Technique Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；
LIU Xingyong	1.Quality Standard and Testing Technique Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；
MI Yanhua	1.Quality Standard and Testing Technique Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；
YIN Benlin	1.Quality Standard and Testing Technique Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；
CHEN Zeli	1.Quality Standard and Testing Technique Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；
WANG Xuejiao	2.Agricultural Economics and Informations Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；
LUO Gang	3.Ning′er Plant Protection and Inspection Station,Pu′er 665199,Yunnan,China；
SONG Yilin	4.Station of Baoshan Agricultural and Mechanizitional Extension,Baoshan 678000,Yunnan,China
LIU Zhenhuan	1.Quality Standard and Testing Technique Research Institute,Yunnnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China；

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