Determination of amentoflavone in Selaginella medicinal herbs from different areas and study on HPLC fingerprint characteristics of Selaginella plants

摘要

Objective To determine the content of amentoflavone in different areas of Selaginella medicinal materials and to establish a method to analyze HPLC fingerprint characterisitics of 10 plants from Selaginella.Method HPLC was applied for establishment of fingerprints,which were used to evaluate and distinguish the different species of Selaginella.Compare their different levels of differences in medicines Selaginella Methods HPLC was used with Agilent zorbax C18 analyticalcolum (4.6 mm×250 mm,5μm).The mobile phase consisted of methanol(A) and phosphoric acid solution (0.1%)in water (B)using a gradient program.The gradient condition elution had the following profile,: 0→17min with methanol 64%(A), 17→45min with methanol 64%→90%(A), 45→55nun with methanol 90%→64%(A).The column temperature was used at room temperature(25C);The flow rate 1.0ml/minDAD detector was set at 330nm for aqcuiring chromatograms.Absorb respectively theabove-mentioned reference solution lOyl and the sample solution 20p.l and then they were drawn into theliquid chromatography. Result I inportant mutual fingerprint peaks were found in the lO plants ofSelagineILa species and 4 peaks can be used a(main frngerprint peaks".The dates of these peaks can usedfor assessment of phylogenetic relation among species and evaluation of quality.According to thecalculation of amentoflavone n(number of plates~3000.Examination of linear relationship:Amentoflavone showed a good linearity in the range of 0.0667-0.1366 hr9.The average recocery was98%and RSD was 0.4%(n=5) .The peak area of the vertical axis, the reference substance of amentotlavonesample size for the abscissa.the standard curve.Regression equation:Y=64490x-0.1639 R=l.Precision Test:measured the average peak area is 4424.5 RSD=1.4%(n=5).Stability Test: measured the average peak areais 4185.8,RSD~.95%.(n=5).Repeatability Test: Calculated for the average content of amentoflavone is1.8%RSD=O.I%(n=5).Conclusion The quantitative of amentotlavone could be as Selaginellatramariscina (Beauv).Spring a quality control reference of Selaginetlaceae.Aamentoflavone indifferent places, quite different in different species of Selaginella The method is accurate and reliable,reproducible, and to be better control the intrinsic quality of Selaginella medicinal materials provide ascientific basis.The fingerprint chromatogram of entire herb constituents of Selaginellaceae Selaginellatramariscina(Beauv.)Spring was developed by HPLC method for the first time.HPLC-performance liquidchromatography paired with photodiode-array detector was analyzed as a method based on the consistentchromatographic features among 10 batches of authentic samples.The HPLC fingerprint of Selaginellatramariscina has been established for the first time.According to the chromatographic condition a goodfingerprint of Selaginella tramariscina was obtained.All chromatographic peaks showed good resolution inthe liquid chromatogram.This work has been done systematically by other scholars.The unique propertiesof this HPLC-DAD fingerprint were successfully applied to analyze and differentiate samples fromdifferent fegions.The resultshowed that a comprehensive quality evaluation of Chinese herb Selaginellatramariscina, which is critical to assess and repressent overall quality.