Development And Application Of Xinfeng Capsule Characteristic Fingerprint By High Performance Liquid Chromatography

Xinfeng capsule, a hospital traditional Chinese herbal formula of The First Affiliated Hospital of Anhui University of Chinese Medicine(Anhui medicine system Z20050062) consists of four herbs, which are astragalus, myotonin, thunder god vine and centipede, and has been used in the treatment of RA about 15 years. Clinical studies have identified that Xinfeng capsule has unique advantages and less side effects as one of the ideal Traditional Chinese Medicine(TCM) on rheumatoid arthritis therapy.Objective:To establish High Performance Liquid Chromatography(HPLC) characteristic fingerprint for Xinfeng capsule, match characteristic peaks with formula herbs,Meanwhile quantitative analysis of the main active ingredients in the preparation,provide a reference for the scientific evaluation of its quality.Methods:1.Consulting literature material, screened the best extraction method of Xinfeng capsule. 2. Study chromatogram conditions, mobile phase, detection wavelength, estab-lish Xinfeng capsule alcohol position fingerprint using HPLC- DAD method. For fingerprint analysis, several peaks were selected as the common peaks, the similarities of 10 batches of Xinfeng capsules were calculated. 3. By the reference of astragalus,myotonin, thunder god vine and centipede, contrast analysis of medicinal materials and preparations, study the correlation between the 4 herbs and preparations.4.Quantitative determination of main active components in Xinfeng Capsule.Results:According to the document retrieval results, summarized the optimal extraction method of Xinfeng Capsule fingerprint was methanol extraction. All separation steps were carried out on an Agilent Eclipse plus C18 column(4.6 mm×250 mm, 5 μm). The mobile phase was composed of 1% phosphoric acid aqueous solution(A) and acetonitrile(B) in gradient elution mode. The flow rate of the mobile phase was kept at1 ml/min and the gradient of acetonitrile(B) was changed as follows: 0-10 min, 5%;10-30 min, 5-15%; 30-55 min, 15-25%; 55-75 min, 25-35%; 75-95 min, 35-45%;95-110 min, 245-65%; 110-130 min, 65-90%; 130-140 min, 90-95%; 140-145 min,95-100%; and 145-155 min, 100%. During fingerprint analysis, the wavelength was set at 210 nm for exhibiting the vast majority of chromatographic peaks. The column temperature was kept at 35℃and the sample injection volume was 10 μl.The similarity index of ten batches Xinfeng capsule was in a range of 0.911-0.990,The main chromatographic peaks from Radix Astragali and Tripterygium wilfordii, and coix seed and centipede in fewer ingredients. The average content of formononetin,calycosin glycoside, calycosin separately for 0.0434 mg/g, 0.1171 mg/g, 0.0717mg/g, according to the calculation of the average content of 80%, request content limit of Xinfeng Capsule for formononetin shall not be less than 0.0347 mg/g,calycosin glycoside shall not be less than 0.0937 mg/g, calycosin shall not be less than0.0573 mg/g.Conclusion:Xinfeng capsule alcohol position characteristic spectrum with good method precision, good repeatability, quantitative analysis method for multi index is reasonable and feasible, and can be used for the quality control of Xinfeng capsule. This method can be used for the quality control of Xinfeng capsule, can offer the reference for the study of other compound preparation.