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.