救必應酸的單體制備及RP—HPLC法測定救必應藥材及精制品中3種活性成分的含量

　　中圖分類號 R284.1 文獻標志碼 A 文章編號 1001-0408（2018）03-0326-04
　　DOI 10.6039/j.issn.1001-0408.2018.03.09
　　摘 要 目的：制備高純度的救必應酸并測定救必應藥材及精制品中3種活性成分紫丁香苷、長梗冬青苷、救必應酸的含量。方法：采用醇提法制備救必應皂苷，以甲醇和氫氧化鈉進一步水解，并經重結晶后得到救必應酸單體。采用反相高效液相色譜（RP-HPLC）法測定救必應藥材及精制品1和精制品2中紫丁香苷、長梗冬青苷、救必應酸的含量。色譜柱為Agilent XDB C18，流動相為水-乙腈（梯度洗脫），柱溫為30 ℃，流速為1.2 mL/min，檢測波長為210 nm，進樣量為10 μL。結果：所制救必應酸純度達99%以上。紫丁香苷、長梗冬青苷、救必應酸進樣量分別在0.18～3.22、0.59～10.36、0.20～3.53 μg范圍內與各自峰面積積分值呈良好線性關系（r均為0.999 9），精密度、穩(wěn)定性（24 h）、重復性試驗的RSD均小于2.0%（n=6～7），平均加樣回收率為96.08%～100.81%（RSD≤1.98%，n=6）。紫丁香苷、長梗冬青苷、救必應酸的含量在救必應藥材中分別為1.61%、7.26%、1.29%（RSD≤1.08%，n=3），在精制品1中分別為0、82.59%、4.18%（RSD≤1.67%，n=3）；在精制品2中分別為0、73.29%、7.41%（RSD≤1.15%，n=3）。結論：所制救必應酸純度高、制備方法簡單且安全環(huán)保。建立的RP-HPLC法簡單、可靠，可滿足快速測定救必應藥材及其精制品含量的要求。
　　關鍵詞 救必應；紫丁香苷；長梗冬青苷；救必應酸；制備；反相高效液相色譜法
　　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

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