| This dissertation includes four part:summary of the related literatures;studies on the chemical constituents of Phyllanthus emblica;studies on the extraction technology of the polyphenol;establishment of methods to control the quality of polyphenol fraction.Part 1:summary of the related literaturesStudies on chemical constituents and pharmacological activities of Phyllanthus emblica L.in recent years were reviewed.83 references were cited.Part 2:studies on the chemical constituents of Phyllanthzts emblica.Several separation and purification methods and determination technology(such as UV,FAB-MS,1HNMR,13CNMR) were used to study the chemical constituents of Phyllanthus emblica.6 polyphenol compounds and 1 flavonoid glycoside were separated,and 6 of them were identified,including gallic acid,ellagic acid,mucic acid 2-O-gallate,1-O-galloyl-β-D-glucose,chebuligic acid,quercetin 3-O-rhamnoside.Part 3:studies on the extraction technology of the polyphenol.The extraction technology of polyphenol fraction of Phyllanthztsemblica L.has been studied.L9(34) orthogonal test has been used to evaluate the best extraction technology,including the best extraction solvent,amount of the solvent,hours and number of times,of polyphenol extraction which was evaluated by the extraction efficiency of total phenol as well as the indicative constituents including诃黎勒酸,gallic acid,mucic acid 2-O-gallate.Finally,the best technology is that using D3 times dosage A2%ethanol,to extract the medical material C3 time,B3 hours every time. The extraction technology described above was tested and verified.The preparation and determination results showed that the established extraction technologies were steady and repeated.Part 4:Establishment of methods to control the quality of polyphenol constituent of phyllanthus emblica.The FeCl3-K3[Fe(CN)6]2 colorimetry was established to determine the total phenol in the polyphenol extractive of Phyllanthus emblica L.The content of total phenol was determined with FeCl3-K3[Fe(CN)6]2 reaction colorimetry,and selected gallic acid as reference substance.Gallic acid had a liner relationship with the absorbance in the range of 0.204～2.040μg/ml and the regression equation was as follows:y=0.5338x+0.0047(r=0.9992).The average recovery for gallic acid was 101.57%,RSD=2.65%(n=6).The FeCl3-K3[Fe(CN)6]2 colorimetry was established to determine the total phenol in the polyphenol extraction of Phyllanthus emblica L.The content of total phenol was determined with FeCl3-K3[Fe(CN)6]2 reaction colorimetry,and selected gallic acid as reference substance.Gallic acid had a liner relationship with the absorbance in the range of 0.204～2.040μg/ml and the regression equation was as follows:y=0.5338x+0.0047(r=0.9992).The average recovery for gallic acid was 101.57%,RSD=2.65%(n=6).After the investigating of several factors,the method was selected as followed:comminutes it into powder which is minute enough to go through 80 mesh cribblel,weighed it about 50mg,adds 50%ethanol in it accurately, weigh it,ultrasonic extract with 100MHz for 30 min,weigh again and then filter, determine the content of total phenol with FeCl3-K3[Fe(CN)6]2 colorimetry;and a casein absorbing method was used to determine the content of tannins.Gallic acid had a liner relationship with the absorbance in the range of 0.204～2.040μg/ml and the regression equation was as follows:y=0.5338x+0.0047(r=0.9992).The content of total phenol of crude drugs was 17.60%,the content of tannins of crude drugs was 7.07%,40.17%to occupy in total phenol.The preparation and determination results showed that the established extraction technologies were steady and repeated.Referring to the method in Pharmacopoeia of China,a tungsten molybdophosphate-casein colorimetric method was used for the determination of tannin in the medical material of Phyllanthus emblica L.with gallic acid as reference substance.Gallic acid had a liner relationship with the absorbance in the range of 1.0072～10.072μg/ml and the regression equation was as follows:y=0.0986x+ 0.0105(r=0.9990).The content of total phenol of crude drugs was 22.67%,the content of tannins of crude drugs was 9.57%,42.25%to occupy in total phenol.The content of total phenol,tannins and the percentage was higher than the FeCl3-K3[Fe(CN)6]2 colorimetry.HPLC methods were established for the determination of three polyphenol,i.e. chebuligic acid,gallic acid,mucic acid 2-O-gallate in the medical material and polyphenol extraction of Phyllanthus emblica L.The linear ranges of chebuligic acid,gallic acid,mucic acid 2-O-gallate were:0.0728～0.364μg,0.1804～0.902μg,0.3872～1.1616μg,respectively.The regression equations of these three polyphenol were as follows:y=2E+09x-489.43,r=0.9996,chebuligic acid;y=2E+09x+51.095, r=0.9996,gallic acid;y= 6E+08x-88530,r=0.9996,mucic acid 2-O-gallate。The average recoveries for three polyphenol were 99.92%,100.06%,99.85%,the RSD were 2.45%,2.35%,2.21%,respectively.Determined the contents of chebuligic acid,gallic acid,mucic acid 2-O-gallate in the polyphenol extraction of Phyllanthus emblica L.under 275nm. |
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