Evaluative Research Of The Quality Of Ganmao Qingre Granule

Ganmao Qingre Granule prescription includes Nepeta ear, Mint, Fangfeng, Bupleurum, Basil leaves, Pueraria, Platycodon grandiflorum, Bitter almond, Angelica dahurica, Kudiding and Phragmites, which has the effect of clearing heat and cold, treatmenting chill cold, headache, fever, body cold, body pain, runny nose, cough, and dry throat.It exerted comprehensive treatment effect based on its containing a variety of chemical constituents. The classic methods applied in the quality control of traditional Chinese medicine was only to measure the content of one component, thus this detection method was not sufficreat to fully reflect its quality. Characteristic chromatogram of traditional Chinese medicine makes a comprehensive evaluation to assess the intrinsic quality of traditional Chinese medicine feasible, and could help us to distinguish the corresponding ownership of its components, therefore it became an effective method for the quality control of Chinese medicine. HPLC, GC and TLC was used to study characteristic chromatogram of Ganmao Qingre Granule, initially indentify three components of corresponding peaks and their categories, and finally provide reference for quality evaluation system of Ganmao Qingre Granule.1HPLC specific chromatogramsTo establish the method of detecting the specific chromatograms method of Ganmao Qingre Granule by HPLC:The samples were separated on Agilent ZORBAX SB C18(4.6mm×250mm5μm) column, with mobile phase consisting of methanol (A) and water (B) and at the flow rate of1.0ml·min-1, the UV detection wavelength was250nm, column temperature was30℃, the gradient elution program:A0â†'15min,5%â†'25%;15â†'55 min,25%â†'70%;55â†'75min,70%â†'95%. At first, we optimized the method and then opplied it in different rastrameats and columns. Finally we preliminarily ideatify the components of common peaks based on the results above with Chromatographic Fingerprint Evaluation System Version2.0.The results show that there were10common peaks, the RSD of the relative retention time in precision, stable and repeatability of experiments were less than0.6%, and the RSD of relative peak area was less than5.0%. The standard specific chromatogram was established, and similarity analysis was carried out. The method is simple, rapid and reproducible, which put forward higher requirements for product quality, and provides help for the quality evaluation of Ganmao Qingre Granule.2GC specific chromatogramsTo establish its characteristic chromatogram with GC. Methods: Agilent HP-5capillary column (30m×320mm,0.25μm) was adopted; injection port temperature was300℃;detector temperature was320℃;Temperature program as follows:the initial column temperature was65℃maintained for5min, then1℃·min-1up to75℃maintained for2min,10℃·min-1up to180℃maintained for10min, finally5℃·min-1up to300℃maintained for10min; the carrier gas was nitrogen and its flow rate was3.8ml·min-1; splitless. To provide experimental evidences for the quality control of Ganmao Qingre Granule and analyze the preliminary correspondence relationship between the peak and a particular ingredient.Results:A total of6peaks, shared by all the batches of Ganmao Qingre Granule, were marked in the characteristic Chromatographic Fingerprint. Precision, stability and repeatability meet the requirements. During the analysis of the similarity, there was minor difference in the categories of ingredients from various batches, but the differences in the contents and ratios of various ingredients were obvious. The separation degree of6peaks was satisfactory in the GC Chromatographic Fingerprint. Pulegone and menthol were detected in most batches and their separation degree met the criteria. These results provided a reference to the quality evaluation of the Ganmao Qingre Granule and also offered useful information for the future quality control in the preparation of Traditional Chinese Medicine Patent Prescription.3TLCTo qualitatively detect herbs in Ganmao Qingre Granule prescription by TLC based on the Pharmacopoeia.4Separate decoction and Merging decoction by simulating the Ganmao Qingre Granule preparation, then to contrast the difference of the characteristic chromatogram with HPLC by two ways, which provide reference for Prescription streamlining.Results:The similarity of the two HPLC chromatography has reached more than0.99, indicating that the prescription was stable, effective.