关防风HPLC特征指纹图谱研究及4个色原酮类成分一测多评法分析研究

Study on fingerprint and multi-component quantitation by one marker method of Radix Saposhnikoviae from northeast

目的:建立指纹图谱及一测多评法测定关防风中4个化学成分含量。方法:采用Agilent ZORBAX Eclipse XDB C18(2)色谱柱(4.6 mm×250 mm,5μm),以甲醇(A)-水(B)为流动相,梯度洗脱(0～10min,20%A→40%A;10～30 min,40%A→65%A;30～40 min,65%A;40～60 min,65%A→80%A;60～70min,80%A),流速1.0 mL·min^(-1),检测波长254 nm,柱温30℃;采用高效液相色谱法测定12批关防风的指纹图谱,建立指纹图谱共有模式。以5-O-甲基维斯阿米醇苷为内参物,建立升麻素、升麻素苷、亥茅酚苷的相对校正因子,并进行含量计算,实现一测多评。同时采用外标法测定12批关防风中4个色原酮类成分的含量,比较计算值与实测值的差异,验证一测多评法的准确性。结果:12批关防风指纹图谱标定了共20个共有峰,指认了其中7个化学成分。12批样品指纹图谱与对照指纹图谱的相似度均大于0.90。建立的相对校正因子重现性良好,采用校正因子计算的含量值与实测值之间无显著差异(P>0.05);通过聚类分析,栽培品和野生药材能有效区分。结论:所建立的方法准确、可行,可用于关防风中4个化学成分的定性定量分析。

Objective:To establish fingerprint and quantitative analysis of multi-components by single marker(QAMS)method for 4 components of Radix Saposhnikoviae from northeast. Methods:The analysis was carried out on an analytical column Agilent ZORBAX Eclipse XDB C18(2)(4.6 mm×250 mm,5 μm)with gradient elution by methanol(A)-water(B)(0-10 min,20%A → 40%A;10-30 min,40%A → 65%A;30-40 min,65%A;40-60 min,65%A → 80%A;60-70 min,80%A),at the detection wavelength of 254 nm and a flow rate of 1.0 mL·min-1;the column temperature was 30 ℃;12 batches of Radix Saposhnikoviae from northeast were determined by HPLC and a common mode of fingerprint has been established. A QAMS method was developed to determine prim-O-glucosylcimifugin,cimifugin,5-O-methylvisammioside and sec-O-glucosylhamaudol. 4’-O-β-Dglucosyl-5-O-methylvisamminol was selected as internal reference;the relative correction factors(RCF)of other three components to the internal reference were calculated. 12 batches of Radix Saposhnikoviae from northeast were determined by both external standard method and QAMS. Results:There were 20 common peaks in fingerprints of 12 batches of Radix Saposhnikoviae from northeast and seven of them were identified. The similarities of fingerprints of 12 batches of samples were above 0.90. The established correcting factors had a good reproducibility.No significant differences(P>0.05)were found between the quantitative results of external standard method and QAMS. The cultivated could be distinguished effectively from wild medicinal materials by cluster analysis.Conclusion:The developed method is accurate,feasible,and can be used for the qualitative and quantitative analysis of Radix Saposhnikoviae from northeast.