Study On Chemical Constituents And Quality Analysis Of Zhuang Medicine Dallergia Hancai Benth

Objective:（1）To study the chemical constituents of ethyl scetate fraction of the Dallergia hancai Benth（2）UV,IR and TLC were used to identify the stems and leaves of Dallergia hancai Benth,and the water content,ash content and extractives of the stems and leaves of Dallergia hancai Benth was determined.（3）To establish the fingerprint of stems and leaves of Dalbergia hancei Benth by HPLC.（4）To determine quercetin and protocatechuic acid in the stems of Dalbergia hancei Benth by HPLC and optimize the extraction technology of total flavonoids from the stems of Dalbergia hancei Benth.Methods:（1）Separation and purification of the constituents of ethyl scetate fraction of the stems of Dalbergia hancei Benth.Which were isolated and purified by repeated silica gel column chromatography,gel column chromatography,and also were recrystallized by different solvents.Their chemical constituents of ethyl acetate were identified by spectral data analysis.（2）According to the Pharmacopoeia of the Peple’s Republic of China（2015 edition）,the stems and leaves of Dalbergia hancei Benth were qualitatively identified by UV,IR and TLC,and the water,ash and extract contents of the stems and leaves of Dalbergia hancei Benth were determined.（3）To establish fingerprint of stems and leaves of Dalbergia hancei Benth collected from different habitats by HPLC.（4）Quercetin and protocatechuic acid in stems of Dalbergia hancei Benth collected from different habitats were determined by HPLC,and extraction technology of total flavonoids from stems of Dalbergia hancei Benth was optimized by central composite design-response surface.Results:（1）Six compounds were isolated and identified from the ethyl acetate fraction from the stems of Dalbergia hancei Benth as:（1）octacosanoic acid,（2）triacontanoic acid,（3）beta-sitosterol,（4）ethyl gallate,（5）protocatechuic aldehyde and（6）quercetin.The above six compounds were separated from Dalbergia hancei Benth for the first time.（2）To determine the identification methods of UV,IR and TLC.To determine the water,ash and extract contents of the stems and leaves of Dalbergia hancei Benth systematically,and determine the detection range tentatively.（3）The fingerprint of stems and leaves of Dalbergia hancei Benth was established by HPLC.Quercetin was used as reference peak in the stems of Dalbergia hancei Benth and rutin as reference peak in the leaves of Dalbergia hancei Benth.All the batches of the sample similarity were over 0.90.The stems and leaves of Dalbergia hancei Benth from different habitat were clustered into 4 groups and 3 groups respectively.（4）Quercetin showed a good linear relationship in a range of 0.2570-1.0210μg and protocatechuic acid showed a good linear relationship in a range of0.2570-1.0210μg.The average recovery of quercetin was 98.10% and the average recovery of protocatechuic acid was 97.93%.The RSD of other methodologies were less than 3%.The quercetin and protocatechuic acid in 10 batches of stems of Dalbergia hancei Benth ranged from39.69μg/g to 62.60μg/g and 13.58μg/g to 32.69μg/g respectively.Central composite design-response surface was used to optimize the extraction technology of total flavonoids from the stems of Dalbergia hancei Benth.The optimum extraction technology was as follow: ratio of liquid to material 59mL·g^-1,extraction tim-e 104 min and the concentration of ethanol 50%.The extraction rate of total flavonoids was 6.60%.Methodologically,rutin showed a good linear relationship in a range of 8.64 μg/ml-43.20μg/ml and the average recovery was 97.6%.The RSD of other methodologies were less than 3%.Conclusion:（1）In this study,the six chemical constituents of ethyl acetate fraction from the stems of Dalbergia hancei Benth were studied,which provided the theoretical basis of chemical constituent in other parts of stems of Dalbergia hancei Benth.（2）A preliminary study was made on the quality analysis of the stems and leaves of Dalbergia hancei Benth.The qualitative identification of the stems and leaves of Dalbergia hancei Benth by UV,IR and TLC and the determination of the water content,ash content and extractives of the stems and leaves of Dalbergia hancei Benth could provide a scientific basis for its quality control and the formulation of quality standards in the future.（3）The HPLC fingerprint of the stems and leaves of Dalbergia hancei Benth was established to provide the relevant theoretical reference for the quality evaluation of Dalbergia hancei Benth.（4）To establish a preliminary method for the determination of quercetin and protocatechuic acid in the stems of Dalbergia hancei Benth by HPLC and optimize the extraction technology of total flavonoids from the stems of Dalbergia hancei Benth,which can be used for quality evaluation of Dalbergia hancei Benth.