救必应酸的单体制备及RP—HPLC法测定救必应药材及精制品中3种活性成分的含量

　　中圖分类号 R284.1 文献标志码 A 文章编号 1001-0408（2018）03-0326-04
　　DOI 10.6039/j.issn.1001-0408.2018.03.09
　　摘要目的：制备高纯度的救必应酸并测定救必应药材及精制品中3种活性成分紫丁香苷、长梗冬青苷、救必应酸的含量。方法：采用醇提法制备救必应皂苷，以甲醇和氢氧化钠进一步水解，并经重结晶后得到救必应酸单体。采用反相高效液相色谱（RP-HPLC）法测定救必应药材及精制品1和精制品2中紫丁香苷、长梗冬青苷、救必应酸的含量。色谱柱为Agilent XDB C18，流动相为水-乙腈（梯度洗脱），柱温为30 ℃，流速为1.2 mL/min，检测波长为210 nm，进样量为10 μL。结果：所制救必应酸纯度达99%以上。紫丁香苷、长梗冬青苷、救必应酸进样量分别在0.18～3.22、0.59～10.36、0.20～3.53 μg范围内与各自峰面积积分值呈良好线性关系（r均为0.999 9），精密度、稳定性（24 h）、重复性试验的RSD均小于2.0%（n=6～7），平均加样回收率为96.08%～100.81%（RSD≤1.98%，n=6）。紫丁香苷、长梗冬青苷、救必应酸的含量在救必应药材中分别为1.61%、7.26%、1.29%（RSD≤1.08%，n=3），在精制品1中分别为0、82.59%、4.18%（RSD≤1.67%，n=3）；在精制品2中分别为0、73.29%、7.41%（RSD≤1.15%，n=3）。结论：所制救必应酸纯度高、制备方法简单且安全环保。建立的RP-HPLC法简单、可靠，可满足快速测定救必应药材及其精制品含量的要求。
　　关键词救必应；紫丁香苷；长梗冬青苷；救必应酸；制备；反相高效液相色谱法
　　ABSTRACT OBJECTIVE： To prepare high-purity rotundic acid， and to determine the contents of syringoside， peduncloside and rotundic acid in Ilicis rotundae and its refined products. METHODS： I. rotundae saponins was prepared by ethanol extraction method and hydrolyzed with methanol and sodium hydroxide； rotundic acid monomer was obtained after recrystallization. The contents of syringoside， peduncloside and rotundic acid in I. rotundae， its refined product 1 and its refined product 2 were determined by RP-HPLC. The determination was performed on an Agilent XDB C18 column with mobile phase consisted of water-acetonitrile （gradient elution） at the flow rate of 1.2 mL/min. The column temperature was 30 ℃， and the detection wavelength was set at 210 nm. The sample size was 10 μL. RESULTS： The purity of rotundic acid exceeded 99%. The linear ranges of syringing， peduncloside and rotundic acid were 0.18-3.22， 0.59-10.36 and 0.20-3.53 μg， respectively （r=0.999 9）. RSDs of precision， stability （24 h） and reproducibility tests were all lower than 2.0% （n=6-7）. The average recoveries were 96.08%-100.81%（RSD≤1.98%，n=6）. The contents of syringoside， peduncloside and rotundic acid in I. rotundae were 1.61%， 7.26%， 1.29% （RSD≤1.08%， n=3）； those of refined product 1 were 0， 82.59%， 4.18% （RSD≤1.67%， n=3）； those of refined product 2 were 0， 73.29%， 7.41% （RSD≤1.15%，n=3）， respectively. CONCLUSIONS： Prepared rotundic acid has high purity and is simple in preparation， safe and environmentally-friendly. RP-HPLC method that established is simple and reliable， and can meet the requirements for rapid analysis of I. rotundae and its refined products.
　　KEYWORDS Ilicis rotundae； Syringoside； Peduncloside； Rotundic acid； Preparation； RP-HPLC