目的 建立基于对照制剂的三萜指纹图谱结合化学计量学分析的方法,测定89批样品以评价灵芝制剂的质量。方法 样品经甲醇超声提取,采用Agilent TC C18(2)色谱柱(4.6 mm×250 mm,5 μm),以乙腈-0.02%磷酸水溶液进行梯度洗脱,检测波长为254 nm,并使用主成分分析和聚类热图分析方法对数据进行统计。结果 建立的指纹图谱共计56个特征峰。样品指纹图谱经与对照制剂比对,发现基本上所有企业都使用赤芝生产灵芝制剂。经主成分分析、聚类热图分析和相似度分析,可将各企业样品分为三类,第一类特征峰种类和峰面积与对照制剂较一致,质量较好;第二类特征峰种类与对照制剂较一致但峰面积偏小,质量一般;第三类与对照制剂差异很大,为问题样品。区分各类样品的强特征峰为峰26、31、24、32和18。结论 该法全面、准确,具有专属性,可用于灵芝制剂三萜成分的整体质量评价。
Abstract
OBJECTIVE To establish a method combining triterpenoids fingerprint with chemometric analysis based on reference drug, and determine 89 batches of samples to evaluate the quality of Ganoderma preparations. METHODS The samples were extracted by ultrasonic with methanol, chromatography was performed on Agilent TC C18(2) column (4.6 mm×250 mm, 5 μm) with gradient elution with acetonitrile and 0.02% phosphoric acid aqueous solution. The detection wavelength was set at 254 nm. Principal component analysis and HCA heatmap were used for data analysis. RESULTS The chromatographic fingerprint has 56 characteristic peaks. By comparing the samples with the reference drug, it was found that Ganoderma preparations were produced using Ganoderma lucidum by almost all manufacturers. Principal component analysis, HCA heatmap analysis and similarity analysis divided the samples into three categories. The first type had characteristic peaks and peak areas which were consistent with the reference drug, and the quality was better. The second type had characteristic peaks which consistent with the reference drug, but the peak areas were smaller and the quality was medium. And the third type was different from the reference drug, which was the problem sample. The strong characteristic peaks for classification of different samples were peak 26, peak 31, peak 24, peak 32 and peak 18. CONCLUSION The method is comprehensive, accurate and specific, and it can be used for comprehensive quality evaluation of triterpenoids in Ganoderma preparations.
关键词
高效液相色谱法 /
灵芝 /
三萜 /
指纹图谱 /
对照制剂
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Key words
high performance liquid chromatography /
Ganoderma /
triterpenoid /
fingerprint /
reference drug
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中图分类号：
R284
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