Study On The Quality Of Different Parts Of Prunella Vulgaris

[Objective]A comparative study on the chemical constituents of different parts of Prunella vulgaris was done to find out the characteristic components of Prunella vulgaris medicinal part, a method was established for content determination of the main active ingredient and characteristic components to provide the basis for the improvement of the quality standard of Prunella vulgaris; The content of moisture and ash in spica Prunellae was determined to investigat the different origin of medicinal materials storage quality and provide reference for safety and efficacy of Prunella vulgaris medicine; The study of HPLC fingerprint for17batches of Prunella vulgaris is to establish a control atlas of Prunella vulgaris and provide the basis for the full establishment of quality standard of Prunella vulgaris.[Methods]1、The study on the chemical constituents of different parts of Prunella vulgaris:(1) The HPLC method was used, chromatography conditions were Agilent Eclipse XDB-C18(4.5mm×300mm,6μm) column with gradient mobile phase of acetonitrile-l.0%acetic acid, UV detection wavelength was300nm with the flow rate of0.8mL/min, the sample injection was10μL(2) With70%ethanol reflux extraction, salviaflaside and rosmarinic acid separated by column chromatography and got the structure of the two substances through the analytical detection of physicochemical properties and pop data; The HPLC method was used to establish a method for determination of both, chromatography conditions were Agilent Eclipse XDB-C18(4.5mm×300mm,6μm) column with gradient mobile phase of acetonitrile(A)-1.0%acetic acid(B) and the gradient was10%A to27%A.UV detection wavelength was320nm with the flow rate of1.2mL/min, the sample injection was10μL(3) Ursolic acid was determined by UPLC.The assay was performed on a ACQUITY UPLC BEH C18(2.3mmx150mm,1.5μm) column and eluted with a mobile phase consisted of methanol-1.0%acetic acid (80%)at a flow rate of0.4ml/min.The detection wavelength was21lnm.The sample injection was1μL2、The content determination of moisture and ash in spica Prunellae: according to water determination method(drying) in Appendix IXH for moisture determination,ash detection in Appendix IXK of the2010edition of "Chinese Pharmacopoeia"for total ash and acid insoluble ash determinations.3、The HPLC method was used to establish a control atlas, chromatography conditions were Agilent Eclipse XDB-C18(4.5mm×300mm,6μm) column with gradient mobile phase of acetonitrile(A)-1.0%acetic acid(B).UV detection wavelength was291nm with the flow rate of0.8mL/min, the sample injection was10μL.[Results]1、The comparison by HPLC fingerprints of different parts of7batches of Prunella vulgaris. indicated that salviaflaside is the characteristic component of Prunella vulgaris medicinal parts.The chromatographic peak is almost not detected in other parts, and the main active ingredient,rosmarinic acid,had the highest content in various parts with the largest peak area.The standard curve was in good linearity over the range of0.0046to0.12μg (r2=0.9996), the recovery of salviaflaside was100.1%and RSD was1.71%. The content of salviaflaside was in the range of0.14%o to0.45%o.The standard curve was in good linearity over the range of0.051to.28μg (r2=0.9993),the recovery of rosmarinic acid was99.96%and RSD was1.08%. The content of rosmarinic acid was in the range of1.63%to4.93%.The standard curve was in good linearity over the range of0.1025to0.41mg (r2=0.9993),the recovery of ursolic acid was99.75%and RSD was1.58%. The content of ursolic acid was in the ranse of0.26%to0.52%. 2、The moisture contents of7batches of spica Prunellae:the highest content was13.80%, the lowest was11.41%and the average was12.63%The total ash contents:the highest was16.79%, the lowest was8.01%and the average was10.44%. The acid insoluble ash contents:the highest was8.16%, the lowest was6.03%and the average was6.98%.3、HPLC fingerprints of17samples of Prunella vulgaris were established.13common peaks were selected as the fingerprint peaks in all samples. Among the obtained fingerprints, most of the detected peaks were separated effectively.17samples had high similarities.[Conclusion]1、The HPLC method is simple, sensitive and accurate,which can be used as an effective method for the quality control standard of Prunella vulgaris. The UPLC method is rapid and accurate and can be used as a quick method for determination of ursolic acid.2、The contents from different origins vary greatly.Chinese medicinal materials on every market should be tested for Standardization to ensure the safety and effectiveness of the medicine quality.3、The HPLC fingerprint method of17batches of Prunella vulgaris is of desirable accuracy and repeatability and can be considered as one of the important evidences for evaluating the quality and varieties identification of Prunella vulgaris.