中药山茱萸炮制前后特征化学成分的分析

目的: 确定可用于分析和鉴别炮制前山萸肉和炮制后酒萸肉的特征成分。建立HPLC法同时测定山茱萸样品中没食子酸、5-羟甲基糠醛(5-HMF)、莫诺苷、当药苷、马钱苷和山茱萸新苷的含量。方法: 采用Phenomenex Gemini C₁₈ 110Å(4.6 mm×250 mm,5 μm)色谱柱,以乙腈(A)-0.3%磷酸水(B)为流动相,梯度洗脱(0~20 min,7%A;20~50 min,7%A→20%A;50~65 min,20%A),流速1 mL · min^-1,检测波长240 nm,柱温35 ℃。将15份山萸肉和10份酒萸肉样品中6种化学成分含量测定数据采用SPSS 10.0软件进行组间t检验统计处理。将酒萸肉中含量较高的4种成分进行雷达图分析。结果: 方法学验证结果显示,没食子酸、5-HMF、莫诺苷、当药苷、马钱苷和山茱萸新苷进样量分别在8.174~653.92 μg(r=0.999 5),48.12~3 849.6 μg(r= 0.999 5),44.92~3 593.6 μg(r=1.000 0),9.088~727.042 μg(r=1.000 0),41.36~3 308.8 μg(r=1.000 0)和8.608~688.64 μg(r=0.999 9)范围内与色谱峰面积呈良好的线性关系;平均加样回收率(n=9)均在97.26~103.1%之间,RSD均小于2.8%。样品测定分析结果显示,山萸肉和酒萸肉样品中,除当药苷含量无显著差异外,其余5种成分没食子酸、5-HMF、莫诺苷、马钱苷、山茱萸新苷含量均有显著差异。炮制品酒萸肉中没食子酸、5-HMF含量高于生品山萸肉,但莫诺苷、马钱苷、山茱萸新苷含量低于山萸肉。雷达图分析结果,马钱苷和没食子酸含量呈相对稳定分布,莫诺苷和5-HMF呈 现相反的变化趋势,莫诺苷含量越低,则5-HMF含量越高。结论: 建立的方法符合方法学验证要求。5-HMF和莫诺苷差异显著,可用于鉴别山萸肉和酒萸肉。

Objective: To determine the characteristic chemical components that can be used to analyze and identify crude and wine processed Fructus Corni.An HPLC method was established for the simultaneous determination of gallic acid, 5-hydroxymethylfufural(5-HMF), morroniside, sweroside, loganin and cornuside in Fructus Corni. Methods: The determination was performed on a Phenomenex Gemini C₁₈ 110Å column(4.6 mm×250 mm, 5 μm)with a gradient elution(0-20 min, 7%A;20-50 min, 7%A→20%A;50-65 min, 20%A)consisting of acetonitrile and 0.3% phosphoric acid aqueous solution.The temperature was set at 35 ℃ with a flow rate of 1 mL · min^-1 and the detective wavelength was set at 240 nm.The content determination results of 6 chemical components from 15 unprocessed samples and 10 wine processed Fructus Corni samples were treated with SPSS 10.0 software by t-test between designed groups.The 4 highest contents in Fructus Corni processed with wine were analyzed by relative plot. Results: The linear ranges of the six components were 8.174-653.92 μg(r=0.999 5), 48.12-3 849.6 μg(r=0.999 5), 44.92-3 593.6 μg(r=1.000 0), 9.088-727.042 μg(r=1.000 0), 41.36-3 308.8 μg(r= 1.000 0)and 8.608-688.64 μg(r=0.999 9), respectively.The average recoveries(n=9)of them were 97.26-103.1% with RSDs of recoveries less than 2.8%.The results showed that the contents of 5 components(gallic acid, 5-HMF, morroniside, loganin, and cornuside)had significant difference other than sweroside between crude and wine processed ones.The wine processed samples had higher contents of gallic acid and 5-HMF than crude samples, however, the contents of morroniside, loganin, and cornuside were lower than those in the crude ones.The relative plot showed that contents of loganin and gallic acid had relatively stable distributions.The 5-HMF and morroniside had inversed relation, namely, the lower content of morroniside was, the higher content of 5-HMF existed. Conclusion: The established method met the methodology validation requirement.The significant difference of 5-HMF and morroniside can be used to identify unprocessed and wine processed Fructus Corni.