HPLC法同时测定3种红景天药材中5种化学成分的含量

　　中圖分类号 R284 文献标志码 A 文章编号 1001-0408（2018）18-2515-05
　　DOI 10.6039/j.issn.1001-0408.2018.18.16
　　摘 要 目的：建立同时测定大花红景天、狭叶红景天和长鞭红景天药材中没食子酸、红景天苷、酪醇、咖啡酸、对香豆酸含量的方法，并比较3种红景天药材中5种化学成分的含量差异。方法：采用高效液相色谱法测定含量，色谱柱为WondaSil C18，流动相为乙腈-0.3%磷酸溶液（梯度洗脱），检测波长为275 nm，柱温为25 ℃，流速为1.0 mL/min，进样量为10 μL；采用GraphPad Prism 5.0软件对含量测定结果进行单因素方差分析和Tukey’s多重比较。结果：没食子酸、红景天苷、酪醇、咖啡酸、对香豆酸的检测进样量线性范围分别为0.020 48～0.614 4 ?g（r=0.999 7）、0.118～3.54 ?g（r=0.999 5）、0.010 84～0.325 2 ?g（r=0.999 7）、0.008 48～0.254 4 ?g（r=0.999 5）、0.004 1～0.123 ?g（r=0.999 5）；精密度、稳定性、重复性试验的RSD均小于2%；加样回收率分别为99.62%～104.92%（RSD=1.68%，n=9）、96.55%～100.69%（RSD=1.67%，n=9）、98.91%～103.39%（RSD=1.48%，n=9）、100.93%～104.98%（RSD=1.37%，n=9）、97.71%～103.89%（RSD=1.99%，n=9）。大花红景天中没食子酸、红景天苷、酪醇和咖啡酸含量均显著高于长鞭红景天，大花红景天中的红景天苷、咖啡酸含量均显著高于狭叶红景天，差异均有统计学意义（P<0.05，P<0.01或P<0.001）。结论：该方法简便可行、结果准确，可用于同时测定大花红景天、狭叶红景天和长鞭红景天药材中没食子酸、红景天苷、酪醇、咖啡酸、对香豆酸的含量。3种常见红景天药材中的化学成分含量存在较大差异，大花红景天中红景天苷的含量最高。
　　关键词 高效液相色谱法；单因素方差分析；Tukey’s多重比较；红景天；没食子酸；红景天苷；酪醇；咖啡酸；对香豆酸；含量
　　ABSTRACT OBJECTIVE： To establish a method for simultaneous determination of gallic acid， salidroside， tyrosol， caffeic acid and p-coumaric acid in Rhodiola crenulata， R. kirilowii and R. fastigiata， and to compare the content difference of 5 chemical components in 3 kinds of R. rosea. METHODS： HPLC method was adopted. The separation was performed on WondaSil C18 column with mobile phase consisted of acetonitrile-0.3% phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 275 nm， and the column temperature was 25 ℃. Sample size was 10 μL. GraphPad Prism 5.0 software was used for single factor variance analysis and Tukey’s multiple comparison of content determination results. RESULTS： The linear ranges of gallic acid， salidroside， tyrosol， caffeic acid and p-coumaric acid were 0.020 48-0.614 4 ?g（r=0.999 7）， 0.118-3.54 ?g（r=0.999 5）， 0.010 84-0.325 2 ?g（r=0.999 7）， 0.008 48-0.254 4 ?g（r=0.999 5） and 0.004 1-0.123 ?g（r=0.999 5）， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2%. The recoveries were 99.62%-104.92%（RSD=1.68%，n=9）， 96.55%-100.69%（RSD=1.67%，n=9）， 98.91%-103.39%（RSD=1.48%，n=9）， 100.93%-104.98%（RSD=1.37%，n=9）， 97.71%-103.89%（RSD=1.99%，n=9）， respectively. The contents of gallic acid， salidroside， tyrosol and caffeic acid in R. crenulata were higher than R. fastigiata； the contents of salidroside and caffeic acid in R. crenulata were significantly higher than R. kirilowii， with statistical significance （P<0.05，P<0.01 or P<0.001）. CONCLUSIONS： The method is simple， feasible and accurate， and can be used for simultaneous determination of gallic acid， salidroside， tyrosol， caffeic acid and p-coumaric acid in R. crenulata， R. kirilowii and R. fastigiata. There was great difference in the contents of chemical components in 3 kinds of common R. rosea， the content of salidroside is the highest in R. crenulata.