基于UPLC-QTOF-MS和多元统计分析的赤芝道地性研究  

Geoherbalism of Ganoderma lucidum:Based on UPLC-QTOF-MS and Multivariate Statistical Analysis

作　　者：张国亮 王晓彤 王汉波 赵建霞 李振宇 俞飞龙 李振皓 ZHANG Guo-liang;WANG Xiao-tong;WANG Han-bo;ZHAO Jian-xia;LI Zhen-yu;YU Fei-long;LI Zhen-hao(Zhejiang Shouxiangu Botanical Drug Institute Co.,Ltd.,Hangzhou 310000,China;Longevity Valley Botanical Co.,Ltd.,Wuyi 321200,China;Zhejiang Key Agricultural Enterprise Institute of Shouxiangu Rare Herb Products,Wuyi 321200,China;Zhejiang Engineering Research Center of Rare Medicinal Plant,Wuyi 321200,China)

机构地区：[1]浙江寿仙谷植物药研究院有限公司,浙江杭州310000 [2]浙江寿仙谷医药股份有限公司,浙江武义321200 [3]寿仙谷珍稀药材产品省级重点农业企业研究院,浙江武义321200 [4]浙江省珍稀植物药工程技术研究中心,浙江武义321200

出　　处：《中国现代中药》2022年第6期979-989,共11页Modern Chinese Medicine

基　　金：浙江省科技计划项目(2019C02100);浙江省重点农业企业研究院项目(2017Y20001);浙江省农业(食用菌)新品种选育重大科技专项(2021C02073)。

摘　　要：目的:研究不同产地、不同栽培方式对赤芝中三萜类成分的影响,为赤芝的道地性研究提供依据。方法:采用超高效液相色谱-四级杆飞行时间质谱法(UPLC-QTOF-MS)对120批赤芝样品进行化学成分分析。将赤芝样品按照浙产/非浙产、浙产段木/非浙产段木、浙产段木/浙产野生、不同发育时期进行模式分类,应用正交偏最小二乘法-判别分析(OPLS-DA)对赤芝样品进行多元统计分析,用变量重要性投影(VIP)值、t检验筛选出组间差异显著的指标性成分。结果:从供试样品中鉴定出13种响应高、分离度好的灵芝酸类成分。筛选发现,浙产赤芝中灵芝酸D、灵芝烯酸D显著高于非浙产赤芝,可作为浙产赤芝的标志性成分;浙产段木赤芝中灵芝酸D、灵芝酸B、灵芝酸C1、灵芝酸G的含量显著高于非浙产段木赤芝;段木赤芝中灵芝酸A等6个成分均显著高于野生赤芝;随着赤芝子实体逐渐成熟(现蕾期→开伞期→成熟期→采摘期),多数灵芝酸含量呈现下降趋势,但部分样品采摘期(喷孢后)灵芝酸含量较成熟期(喷孢前)高,值得进一步研究。结论:浙产与非浙产赤芝成分差异较明显,且浙产段木赤芝与非浙产段木赤芝成分也存在明显差异,可能与生态环境、种植方式有关。Objective:To explore the influence of cultivation method and producing area on triterpenoids in Ganoderma lucidum and thereby to lay a basis for studying the geoherbalism of G.lucidum.Methods:Ultra-performance liquid chromatography coupled to quadrupole time of flight mass spectrometry(UPLC-QTOF-MS) was employed for chemical identification of 120 batches of G.lucidum samples.To be specific,samples from Zhejiang and other producing areas,samples cultivated with wood log from Zhejiang and other areas,wild samples from Zhejiang and samples cultivated with wood log from Zhejiang,and samples at four different developmental stages were respectively compared.Orthogonal partial least squares discriminant analysis(OPLS-DA) was used for multivariate statistical analysis of the 120 batches of samples.Components with significant difference between groups were screened based on variable importance in projection(VIP) value and t test.Results:A total of13 compounds with high response and satisfying separation were identified from the samples.Content of ganoderic acid D and ganoderenic acid D was significantly higher in the samples from Zhejiang than that in samples from other producing areas,and thus the two can be used as chemical markers of G.lucidum from Zhejiang.The content of ganoderic acid D,ganoderic acid B,ganoderic acid C1and ganoderic acid G in samples cultivated with wood log from Zhejiang was significantly higher than that in samples cultivated with wood log from other places.Six compounds including ganoderic acid A showed higher content in samples cultivated with wood log from Zhejiang than in wild samples from Zhejiang.Moreover,as the fruiting body gradually matured(budding stage→cap growth stage→mature stage→harvest stage),most of the ganoderic acids showed a downward trend,but some samples had higher level of ganoderic acids at the harvest stage(after spore ejection) than at mature stage(before spore ejection),which was worthy further study.Conclusion:Significant difference in components was found between

关 键 词：赤芝 道地性 超高效液相色谱-四级杆飞行时间质谱法 正交偏最小二乘法-判别分析 灵芝酸D 灵芝烯酸D 

分 类 号：R284[医药卫生—中药学]