目的 使用赤芝对照提取物(CZERS)、紫芝对照提取物(ZZERS)对所收集18批次灵芝药材进行高效薄层色谱(HPTLC)以及高效液相色谱(HPLC)指纹图谱分析。方法 使用HPTLC指纹图谱方法,高效硅胶预制板,以氯仿-乙腈-甲醇-甲酸=13∶2∶0.5∶0.5 3次展开和环己烷-乙酸乙酯-甲醇-甲酸=15∶5∶0.5∶0.5 2次展开的方法分别分析灵芝三萜酸类和甾醇类成分。使用HPLC指纹图谱方法,Kromasil 100-5 C18柱(4.6 mm×250 mm,5 μm);流动相:A-乙腈,B-0.02%磷酸;梯度洗脱程序:0~40 min,29%→33%A,40~70 min,33%→65%A,70~105 min,65%→100%A;105~120 min,100%A;检测波长244 nm(DAD检测器);流速1.0 mL·min-1;进样量10 μL;柱温25 ℃。结果 使用对照提取物(ERS)和指纹图谱分析方法,可以区分赤芝,无柄灵芝和紫芝。不同种类和生长方式的灵芝成分存在差异。结论 灵芝品种繁多,分野生和人工培养,而人工培养的培养基又有不同,导致不同灵芝个体间成分存在差异。使用特定品种的ERS和指纹图谱分析方法更加适合灵芝多成分整体质量控制的需求。
Abstract
OBJECTIVE To carry out HPTLC and HPLC fingerprint analysis of 18 batches of Ganoderma samples using two kinds of reference substance of Ganoderma extract, G. lucidum Extract Reference Substance(CZERS) and G. sinense Extract Reference Substance(ZZERS). METHODS HPTLC Fingerprint was used to analyze triterpene acids and sterols in Ganoderma with chloroform-acetonitrile-methanol-formic acid (13∶2∶0.5∶0.5, develop 3 times) and cyclohexane-ethyl acetate-methanol-formic acid (15∶5∶0.5∶0.5, develop 2 times) respectively. HPLC Fingerprint analysis was conducted using Kromasil 100-5 C18 column (4.6 mm×250 mm, 5 μm) kept at 25 ℃. Mobile phase A was acetonitrile and B was 0.02% phosphoric acid; gradient elution procedure was as follows: 0-40 min, 29%→33% A; 40-70 min, 33%→65%A; 70-105 min, 65%→100%A; 105-120 min, 100% A; flow rate was 1.0 mL·min-1. DAD detector was adopted with detection wavelength set at 244 nm. The injection volume was 10 μL. RESULTS By using ERS and fingerprint analysis, G. lucidum, G. sessile and G. lucidum could be distinguished. The components of G. lucidum in different species and growth patterns were different. CONCLUSION There are many varieties of G. lucidum, which can be divided into wild and artificial cultures, and the culture media of artificial culture are different, which leads to the difference of individual components of different G. lucidum. Fingerprint analysis based on ERS of specific varieties are more suitable for the overall quality control of G. lucidum.
关键词
对照提取物 /
赤芝对照提取物 /
紫芝对照提取物 /
HPTLC指纹图谱 /
HPLC指纹图谱 /
三萜酸 /
多次展开
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Key words
G. lucidum Extract Reference Substance /
G. sinense Extract Reference Substance /
HPTLC fingerprint /
HPLC fingerprint /
triterpene acid /
multiple development
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中图分类号：
R284
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参考文献
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脚注
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基金
广东省中医药局名优中成药二次开发项目资助(20174007)
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