福建灵芝高效薄层色谱鉴别及其在溯源中的应用

Identification of Ganoderma lucidum from Fujian by high performance thin layer chromatography and its application in traceability

目的本研究旨在建立福建灵芝高效薄层色谱方法,并对比福建产区与其他产区薄层的异同点,为灵芝药材在薄层溯源中的应用提供实验依据。方法采用Merk HPTLC Silica gel 60预制板,氯仿-乙腈-甲醇-甲酸(13∶1.5∶0.2∶0.2)为展开剂,3%硫酸乙酸乙酯溶液显色,在紫外366 nm下检视,采用对照品比对鉴别薄层色谱条带。使用斑点条带数字化表征,对不同产区样品进行聚类分析。结果高效薄层色谱共分离灵芝药材中17个条带,用对照品比对鉴定了其中11个成分,包括灵芝烯酸C(Rf 5=0.31)、灵芝酸C2(Rf_(6)=0.33)、灵芝酸I(Rf_(7)=0.35)、灵芝酸G(Rf_(8)=0.41)、灵芝酸A(Rf_(9)=0.44)、灵芝酸B(Rf_(10)=0.46)、灵芝内酯B(Rf_(11)=0.53)、3β,7β,15β-三羟基-11,15-二羰基-羊毛甾烷-8-烯-24→20内酯(Rf_(12)=0.56)、灵芝烯酸D(Rf_(13)=0.61)、灵芝酸D(Rf_(14)=0.63)和20(21)-脱氢赤芝酸A(Rf_(15)=0.65)。福建灵芝共鉴别10个条带,其中未知成分(Rf_(1)=0.14;Rf_(2)=0.17)、灵芝酸C2和灵芝烯酸D 4个条带是其差异标志物。不同产区灵芝在薄层特征图谱上存有差异。聚类分析当欧式距离为25时,灵芝样本可以被区分为福建产区和其他产区两类。结论建立的高效薄层图谱方法简便、特征明显,可应用于福建灵芝的质量追溯,可为福建灵芝的道地追溯提供一种新的高效薄层色谱技术手段。

Objective To establish the high performance thin layer chromatography(HPTLC)method of Ganoderma lucidum from Fujian,to identify differences between Fujian and other production areas,which provided an experimental basis for the application to Ganoderma lucidum traceability.Methods HPTLC was conducted on Merk silica gel 60 plate,with the TLC solvent system of chloroform-acetonitrile-methanol-formic acid(13∶1.5∶0.2∶0.2).3%sulfuric in ethyl acetate as color developing agent,which was observed under ultraviolet lamp at 366 nm.Cluster analysis was performed on samples from different production areas using the digital characterization of spot bands.Results A total of_(1)7 components were separated by HPTLC,11 components were confirmed by reference substances,including ganoderenic acid C(Rf 5=0.31),ganoderic acid C2(Rf_(6)=0.33),ganoderic acid I(Rf_(7)=0.35),ganoderic acid G(Rf_(8)=0.41),ganoderic acid A(Rf_(9)=0.44),ganoderic acid B(Rf_(10)=0.46),ganolactone B(Rf_(11)=0.53),3β,7β,15β-trihydroxy-11,15-dicarbonyl-lanostane-8-en-24→20 lactone(Rf_(12)=0.56),ganoderenic acid D(Rf_(13)=0.61),ganoderic acid D(Rf_(14)=0.63)and 20(21)-dehydrolucidenic acid A(Rf_(15)=0.65).A total of_(10)bands were identified from Ganoderma lucidum in Fujian.Four bands components(Rf_(1)=0.14,Rf_(2)=0.17),ganoderic acid C2 and ganoderenic acid D(Rf_(14)=0.63)were the differential components between Fujian and other different producing areas by HPTLC.Cluster analysis divided the Ganoderma lucidum into 2 groups at 25 euclidean distance.Conclusion HPTLC method is simple and has obvious characteristics,could be applied to the quality traceability of Fujian Ganoderma lucidum,and providing a new HPTLC technique for the authentic traceability of Fujian Ganoderma lucidum.