ABSTRACT
Objective: To establish the composition-activity relationship (CAR) model based on study of chemical composition and relating proliferation inhibitory rate of Corydalis soxicola and recognize the anti-hepatic fibrosis active compounds of C. soxicola. Methods: Nine orthogonal C. soxicola extract were analyzed by HPLC, and 21 characteristic peaks were profiled. Anti-hepatic fibrosis activity was investigated by MTT assays on HSC-T6, and the potential active components were identified by scores plot and variable importance in projection (VIP) values by means of orthogonal partial least squares (OPLS) analysis, and the activities of identified components were verified by MTT and flow cytometry. LDH kit was used to detect the effect of dehydrocavidine, palmatine, and berberine on LDH activity. Results: The results showed that six peaks including peaks 13, 14, 16, 18, 19, and 20 were significantly related to anti-hepatic fibrosis activity among nine orthogonal extract from C. soxicola. Peaks 18, 19, and 20 were characterized as dehydrocavidine, palmatine, and berberine, respectively. MTT assay showed that dehydrocavidine, palmatine, and berberine with various concentration significantly inhibited the proliferation of HSC-T6 cells. It was also found in flow cytometry that the apoptotic rates of dehydrocavidine, palmatine, and berberine (0.10 mg/mL) on HSC-T6 were 42.12%, 42.22%, and 36.73%, respectively, which were obviously higher than that of control (1.69%, P < 0.01). No obvious cytotoxic effect was found when the final concentration of dehydrocavidine, palmatine, and berberine were less than 0.15, 0.10, and 0.10 mg/mL, respectively. Conclusion: In this study, it was for the first time found that dehydrocavidine, palmatine, and berberine in C. soxicola extract can effectively inhibit the proliferation and induce apoptosis of HSC-T6 based on composition-activity relationship, and there was no obvious cytotoxic effect of the effective concentration, which showed that they may be the active components with potential anti-hepatic fibrosis effect and have a certain security in the application in C. soxicola. At the same time, it also suggests that the research ideas based on CAR can provide an effective method for the identification of active components in natural plants.
ABSTRACT
OBJECTIVE:To establish the quality standard of Corydalis tomentella. METHODS:The medicinal material was identified in respects of property,microscopic characteristics and TLC. The contents of moisture,total ash and water soluble ex-tract were determined. The content of dehydrocavidine was determined by HPLC. The determination was performed on Agilent C18 column with mobile phase consisted of acetonitrile-phosphate buffer solution(containing 20 mmol/L monopotassium phosphate,10 mmol/L diethylamine,0.1% phosphoric acid)(28:72,V/V)at the flow rate of 1.0 mL/min. The detection wavelength was set at 347 nm,and the column temperature was 35 ℃. The sample size was 10 μL. RESULTS:The original plant is perennial herbs. The me-dicinal material often shrank into a ball. The main root was in conical shape;the rhizomes and roots were distinctly chapped;leaves curled and broken,and flowers were yellowish-white. The pollen grains were round-like in shape;square and columnar crys-tals were found;there were a large number of nonglandular hairs. The bordered pit,threaded and reticulate catheter were found;wood fiber was also found. TLC spots were clear and well-separated. The content of moisture were 7.5%-18.5%,total ash were 20.5%-26.2%,and extract were 29.9%-46.4%. The linear range of dehydrocavidine were 0.04008-2.4048 μg(r=0.9999);RSDs of precision,stability and reproducibility tests were lower than 2.0%. The recoveries were 95.6%-102.5%(RSD=2.3%,n=9). CONCLUSIONS:The established standard can be used for quality evaluation of C. tomentella.
ABSTRACT
OBJECTIVE: To establish a method of HPLC fingerprint to determine the contents of primary alkaloids in Corydalis saxicola Bunting cells. METHODS: The determination was performed on a Dikma Diamonsil C18 column (4.6 mm×250 mm, 5 μm). The mobile phase was acetonitrile and water (40:60) supplemented with 3.4 g·L-1 KH2PO4 and 1.7 g·L-1 SDS. The chromatogram was monitored at 345 nm. The flow rate was 1 mL·min-1, and the column temperature was kept at 25°C. RESULTS: The linear regression equations for dehydrocavidine and berberine wereY=3645.2X+47.61 (r=0.9989) and Y=4172.7X+73.52 (r=0.9992), and the average recoveries were 97.9% and 98.2%, respectively. The intra- and inter-day RSDs of these alkaloids were lower than 3%. CONCLUSION: An effective method with good precision was established for the determination of the alkaloids in cultured C. saxicolacells. The Results of resemblances evaluation of the fingerprints were satisfactory. This method can be used as the quality and quantity control of C. saxicola cells.