Study On Quality Control Of Bawei Longzuan Granules

Objective:To establish a TLC identification method of Bawei Longzuan granules for oddalia asiatica,Zanthoxylum nitidum,Orientvine,Sargentodoxa cuneate,Ficus hirta and Bauhinia championii.To establish a HPLC-UV method for the simultaneous determination of the contents of seven components（Gallic acid、Protocatechuic acid、Sinomenine、Salidroside、（+）-Catechin、Hesperidin、Psoralen）in Bawei Longzuan granules and to determine the fingerprint of Bawei Longzuan granules.This study aims to further improve the quality evaluation system and quality control level of Bawei Longzuan Granules,and to provide experimental foundation and scientific basis for the establishment of a more comprehensive and objective quality control standard of Bawei Longzuan Granules.Methods:（1）There chloroform-methanol-concentrated ammonia solution（30:1:0.1）,cyclohexane-ethyl acetate-methanol-isopropanol-concentrated ammonia solution（20:5:3:1:0.1）,chloroform-acetone-formic acid（16:5:0.1）,xylene-ethyl acetate-methanol-water（2:4:2:1）,n-hexane-ethyl acetate（8:2）,and petroleum ether（60～90℃）-chloroform-ethyl acetate-methanol（7.5:4:1.5:0.75）were used as a developing agent,Qualitative analyses of six effective components of Toddalia asiatica;Zanthoxylum nitidum;Orientvine;Sargentodoxa cuneata;Ficus hirta Vahl and Bauhinia championii（Benth.）Benth were carried out,and the specificity and durability of the established method were investigated by thin layer chromatography（TLC）.（2）The HPLC-UV was used to determine seven components（Gallic acid、Protocatechuic acid、Sinomenine、Salidroside、（+）-Catechin、Hesperidin、Psoralen）and fingerprint of Bawei Longzuan granules.The contents were determined by HPLC method on Phenomenex Gemini C₁₈column（4.6 mm×250 mm,5μm）,and the detection wavelengths were set at 270 and 245 nm（0～50min:270nm,50～60min:245nm）,gradient elution,with Acetonitrile and 0.1%phosphoric acid as the mobile phase;the column temperature was30℃,the flow rate was 1.0 m L/min;Results:（1）TLC was use to distinguish six effective components of Toddalia asiatica;Zanthoxylum nitidum;Orientvine;Sargentodoxa cuneata;Ficus hirta Vahl and Bauhinia championii（Benth.）Benth in Bawei Longzuan granules.The results showed that the established TLC identification method had strong specificity,the spots were clearly visible,and the negative control had no interference,which could be used as a qualitative identification method for Bawei Longzuan granules.（2）The HPLC-UV method was use to establish simultaneously determine the content of seven components of gallic acid、protocatechuic acid、sinomenine、salidroside、（+）-catechin、hesperidin、psoralen.And be used for the determination of the characteristic fingerprint of Bawei Longzuan granules and showed the quantitative results as follows:the calibration curves of each solvent showed a good linear relation between the logarithm of peak area and the logarithm of sample size.The linear range of gallic acid,protocatechuic acid,sinomenine,salidroside,（+）-catechin、hesperidin,psoralen 0.2784～0.9744μg,0.0348～0.1218μg,0.8184～2.8644μg,1.0256～3.5896μg,0.4016～1.4056μg,0.0972～0.3402μg,0.0284～0.0994μg;The average recovery were 104.1%、102.1%、102.5%、99.6%、103.2%、99.2%、98.5%,RSD were 1.3%、1.0%、1.4%、1.7%、1.4%、0.92%、1.3%（n=6）.The fingerprints of 10 batches were determined,chromatographic peaks of sinomenine were used as reference peaks to establish a common pattern of fingerprints of Bawei Longzuan granules.By analyzing the fingerprints of 10 batches of Bawei Longzuan granules,a total of 12 common peaks were generated on the control maps,and comparing with the drug maps and comparing with the HPLC and UV spectra of the reference materials,and 7 common peaks were identified.The chemical composition determines the attribution of each common peak.Comparing and analyzing the fingerprint data of 10 batches of Bawei Longzuan granules,and referencing standard to calculate the degree of similarity,the results showed that the similarity is up to 0.989.Conclusion:（1）The TLC method of Identification was established,and can be used to identify 6 kinds of medicinal materials in Bawei Longzuan Granules.his method is simple and special in operation,the results are stable with a good repeatability and showed no interference.These can be well applied to control the quality of Bawei Longzuan Granules.（2）The fingerprint of Bawei Longzuan Granules in this study was combined with the determination method of the main components can be qualitative and quantitative.The fingerprints of Bawei Longzuan Granules have established strong specificity and pertinence.The results of the methodological investigation showed that the method has good repeatability and stability,and the content determination results are accurate.The method provides a scientific basis for the establishment of the quality control method of Bawei Longzuan Granules,and provides an experimental basis and scientific basis for the establishment of a more objective and effective quality standard of Bawei Longzuan Granules.