Evaluation Of Safety & Quality Of Effervescent Adherent Suppository Of Chinese Medicinal Herb "Gong Dejian"

The research group prepared the Chinese medicinal herb compound "Gong Dejian", an effervescent adhesive suppository which is applied in the treatment of bovine endometritis according to the pharmacological action of Chinese medicinal herb. In this research, in order to effectively control its quality, improve the safety of its application and provide the scientific basis for its large-scale production, the bacteriostatic effect in vitro and quality standard were explored, and the safety of the preparation was evaluated using modern detection techniques.The quality of the suppository was evaluated based on the China Veterinary Pharmacopoeia (2010 edition). The suppository mainly contains six kinds of Chinese medicinal herbs:Coptis, Scutellaria, Motherwort, Gardenia, Phellendron, and Betel nut. Tartaric acid and sodium bicarbonate were used as the the acid-base source, polyethylene glycol (PEG-4000 and PEG-6000) as the stroma, hydroxypropyl cellulose (HPC) as the adhesion agent, microcrystalline cellulose as the filler. Effervescent adhesive suppository of "Gong Dejian" was successfully manufactured with modern new pharmaceutical techniques, the weight variation, pH and foaming capacity of the preparation were detected. The results showed that the weight variation was within 5%, average pH value was 4.50±0.050, and average foaming capacity value was 23±3.0 mL. The results were in accordance with China Veterinary Pharmacopoeia(2010 edition). Thin-layer chromatography(TLC) methods of identification of Coptis, Scutellaria, Phellodendron, Gardenia, Motherwort and Betel nut were established. The results showed that the colour spots of the sample corresponded with that of the standard medicines while the negative control displayed no interference on the chromatograph. These methods were verified to be simple, convenient, sensitive, specific and repetitive. High performance liquid chromatographic (HPLC) method was established to determine the three active components of Berberine hydrochloride, Gardenia, and Baicalin, The methods underwent testing. Chromatographic conditions include:Wondasil C18 for Herbal Medicine(5 μm,4.6 X 250 mm) chromatographic column, acetonitrile (A)-0.2% phosphoric acid solution (B) as the mobile phase, gradient elution (0-12 min,5%A-18%A; 12～20 min,18%A～21%A; 20～30 min,21%A～25%A; 30～43 min,25%A～30%A), column temperature 30℃, detection wavelength 240 nm, sampling volume 10 μL, current speed 1.0 mL/min; The results showed that the sampling size of Geniposide, Berberine hydrochloride and Baicalin in 0.1～0.5 μg (r=0.9972),0.1～0.5 μg (r=0.9978) and 0.15～0.75 μg (r=0.9957) respectively reflected good linear relations with peak area. The average recovery rate was 97.99%(RSD%=0.19),98.23%(RSD%=0.19),101.01%(RSD%=0.25) respectively. These results showed that the methods were good at instrument precision, stability, reproducibility, specificity and were suitable for quality control of "Gong Dejian" effervescent adhesive suppository.Bacteriostatic test of "Gong Dejiair" effervescent adhesive suppository on Staphylococcus aureus, Escherichia coli and Streptococcus were conducted in vitro. The medicine was double diluted to 0.1,0.05,0.025,0.0125,0.006,0.003,0.0015,0.00075,0 g/mL and the medicine mediums were made. The inoculated concentration of the bacteria was 104cfu/mL. The results showed that the MIC of "Gong Dejian" effervescent adhesive suppository on Staphylococcus aureus, Escherichia coli and Streptococcus was 1.5,6 and 25 mg/mL respectively.The safety evaluation of "Gong Dejian" effervescent adhesive suppository were as follows:In the eye irritation test of rabbits, medicine was introduced into the left as an irritant whereas the right eye being the control. Results showed that none of the rabbits displayed any irritation reactions. In the vaginal mucosa irritation test of rabbits, after continuous intravaginal administration of medicine for 7 days, the rabbits’physiological activities were observed, and then the rabbits were necropsied and the vaginal tissue examined. Pathological sections were made. Results showed "Gong Dejian" effervescent adhesive suppository had no stimulation to the vaginal mucosa of rabbits. In the cutaneous hypersensitivity test of rabbits, none of the rabbits in the group of "Gong Dejian" nor physiological saline group showed any allergic symptoms. All of the positive control group of 2,4-dinitrochlorobenzene showed allergic symptoms, which indicates "Gong Dejian" effervescent adhesive suppository could not induce cutaneous hypersensitivity on rabbits.