| Yinlian capsule, consisted of L. japonica Thunb, T. chinensis Bunge, and other medicinal materials is one of traditional Chinese medicine recipe for clearing away heat and toxic materials, relieving inflammation and alleviating pain. The method of preparation and quality control, antibacterial and anti-inflammatory activity of Yinlian capsule were studied in this dissertation.The extraction progress for Yinlian extracts from medicinal material was investigated by single-factor and orthogonal experiment methods with the content of the total flavonoids as index. Four factors were studied in this experiment, including the times of extraction, time of reflux, solvent kinds and consumption. The result showed that the optimal extracting condition was 60% ethanol consummated 12 times the amount of material, and three times for one hour each time.The purification progress for Yinlian extacts was established with macroporous adsorption resin. The static and dynamic adsorption tests were employed to investigate effects of separation and purification for the total flavonoids from Yinlian extracts with the recovery rate of the total flavonoids as index. The result showed that the static adsorption capacity of HPD-300 type resin was 37.0 mg·g-1, the static elution ratio was 84.8%. And the optimal purifying condition was as follows:The Yinlian extracts solution of 30.0 mL (the total flavonoids 0.490 mg-mL-1) was loaded for 2 times on the resin column (2×20 cm) with a speed of 4 times volume of the resin-h-1. The impurity was washed away with 2 times volume of the resin waters, and the speed of 12 times volume of the resin-h"1. And then, the totle flavonoids were eluated with 40% ethanol. And the eluant volume was 6 times volume of the resin. The eluting rate was 12 times volume of the resin-h"1. Compared with DM301, D101, DA201 and AB-8, The resin HPD-300 is better for using for separation and purification the total flavonoids from Yinlian extracts. The recovery of flavonoids was more than 85%.The mold process of Yinlian capsule was researched by determination of the angle of repose, hygroscopic rate, and bulk density. The result showed that the optimal mold condition was as follows:the soft material, consisted of Yinlian extract, lactose, microcrystalline cellulose(52:24:24), was screened with 20 mesh for palletizing with 80% ethanol as wetting agent, and dried under 60℃. The secondary screens removed the fines with 60 mesh. And grains during 20-60 mesh were filled into No.0 capsule. The Yinlian capsule was maken.An UV method was established to determine the content of the total flavonoids. The linear ranges was 0.0106-0.0634 mg-mL-1 (r=0.9997). The average recovery (n=6) was 98.8% and with RSD of 2.7%. The content range of herbs and Yinlian extrants was 109.7-113.2 mg·g-1,524.6-539.7 mg-g-1, respectively. This method is simple, fast, stable, and fulfills quantitative analysis requirements.To develop an RP-HPLC method for simultaneous determining chlorogcnic acid, orientin, vitexin, linarin and luteolin in Yinlian extract. The linear ranges of chlorogcnic acid, orientin, vitexin, linarin and luteolin were 0.290-11.6 ug-mL-1 (r=0.9998),0.350-14.0μg-mL-1 (r=0.999 7),0.320-12.8μg-mL-1 (r=0.9997),0.280-11.2μg-mL-1 (r=0.9995),0.110-4.40μg-mL-1 (r= 0.9999), respectively. The average recoveries (n=6) of chlorogcnic acid, orientin, vitexin, linarin and luteolin was 99.4%,105.6%,102.4%,103.2%,92.4% and with RSD of 2.6%,2.1%,2.8%, 1.4%,1.5%, respectively. The content ranges of chlorogcnic acid, orientin, vitexin, linarin and luteolin were 28.5-31.4,39.2-41.8,10.8-11.5,39.8-42.3,2.40-2.58 mg·g-1, respectively. Though methodology verification, this method fulfils quantitative analysis requirements.A quality control method of Yinlian capsule for identifying and determining chlorogcnic acid, orientin, and linarin was established by TLC and RP-HPLC. The sepetation of orientin and linarin were achieved on polyamide thin-layer plate, with mobile phase of 36% acetic acid and ethylacetate:butanone:formic acid:water (5:3:1:1). The linear ranges of chlorogcnic acid, orientin, and linarin were 2.50-40.0μg-mL-1 (r=0.9998),2.70-43.2μg-mL-1 (r= 0.9996),4.64-74.2μg-mL-1 (r=0.9998), respectively. The average recoveries (n=6) of chlorogcnic acid, orientin, and linarin was 95.6%,102.5%,101.6% with RSD of 1.8%,2.4%,2.2%, respectively. The content ranges of chlorogcnic acid, orientin, and linarin were 2.74-2.82,7.36-7.62,12.7-13.3 mg·capsule-1. This method is fast, simple, reproducible, suitable as quality standards for Yinlian capsule.To establish a method for studying the antibacterial activity of yinlian extracts by Drug-containing plate and Cup-plate. Compared with the short-Bacillus and E. Coli., The of antibacterial effect of yinlian extracts on the Staphylococcus aureus is found more sensitive, and the MIC of the total flavonoids in yinlian extracts to Staphylococcus aureus is 6.58 mg-mL-1.The anti-inflammatory activity of Yinlian capsule was investigated by xylene-induced mouse ear edema model. The experimental data indicated that Yinlian capsule had significant inhibitory effects on edema in the ears of mice induced by xylene (P< 0.01). The ear oedema of high, medium and low groups was 15.9±3.93 mg,20.4±5.74 mg,24.1±4.03 mg, respectively. This experiment provides pharmacological basis for development and utilization for Yinlian capsule. |
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