石榴皮抑菌软膏的研制
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摘要
目的:通过对石榴皮抑菌软膏制备工艺、体外抑菌实验、质量标准、稳定性实验和皮肤用药安全性实验等方面的研究,为研制一种新的软膏制剂提供科学依据。方法:1)运用离心实验、耐热和耐寒实验筛选出最佳软膏基质处方。2)进行体外抑菌实验追踪制备工艺。3)用TLC法对石榴皮抑菌软膏进行定性鉴别:采用UV-Vis法进行含量测定。4)进行中试实验,确定工艺路线,依照05版《中国药典》进行检测。5)进行三个月的稳定性考察。6)皮肤用药安全性实验观察对新西兰兔急性毒性和刺激性,对豚鼠的过敏性反应。结果:1)采用耐热、耐寒实验和离心实验考察软膏稳定性,结果均无油水分离现象。2)纸片法和试管稀释法测得石榴皮提取液、浓缩液、冷干粉末及软膏对金黄色葡萄球菌、大肠埃希菌的抑菌圈直径分别在23.5～11.6mm之间,MIC在2500μg/ml～78.1μg/ml之间,MBC在5000μg/ml～156.3μg/ml之间。3)TLC法所得色谱图特征斑点清晰,斑点分离度好;通过分析方法验证,UV-Vis法满足软膏含量测定要求。。4)中试产品质量达到质量标准的要求,检查均符合有关规定。5)经过三个月加速试验稳定性考察样品性状、鉴别、装量、卫生学检查和含量测定等各项指标均无明显变化。6)软膏未产生急性毒性反应;一次或多次给药对动物正常皮肤和破损皮肤均无刺激;致敏后使用,也无过敏反应。结论:1)所制得的5%石榴皮抑菌软膏膏体均匀、细腻,易涂布并无刺激性。2)石榴皮提取液、浓缩液、冷干粉末及软膏均具有较好的体外抑菌活性。3)TLC鉴别检出软膏中石榴皮特征斑点。暂定:软膏的含量限度为没食子酸的含量不得少于0.0667g/g～0.1001g/g,其他指标应符合05版药典的有关要求。4)中试结果表明,该工艺可放大生产,且化学成分转移率较高,工艺合理。5)考察软膏在40℃、相对湿度75%的条件下存放3个月的稳定性,质量基本稳定。6)软膏未产生急性毒性、刺激性和过敏性反应。
Objective: To study the preparation process of pomegranate rind bacteriostatic ointment, antibacterial experiment in vitro, quality standard, stability testing and local application safety test in order to go on further clinic research.Methods: 1) Selected the best prescription ointment matrix with centrifugal test, heat resistance and cold resistance test. 2) The ointment was identified by TLC. The content of the ointment was determined by UV-Vis. 3) The bacteriostasis test in vitro tracked techn ic. 4) The final craft technology was established by mid-experiment. The ointment was determined by relevant rules of ointment according to Chinese pharmacopoeia. 5) Its stability was observed during three months. 6) Local application safety test was observed.It was observed whether the ointment could induce acute toxicity irritation and the allery reaction.Results: 1) To observe the stability of the ointment by cold、hot and centrifuged laboratory experiments. The result showed the ointment was stable. 2) The ointment was identified by TLC. The result showed that the chromatogram of TLC was clear and the resolution was well. After that the quality standard of the ointment was finished. The content of gallic acid in the ointment was determined by UV-Vis, and the average recovery was 99.92% and RSD was 1.02%. 3) Kirby– Bauer method and the tube dilution method were employed for bacteriostasis test in vitro of extract, concentrated liquid, freeze-drying power and ointment of pomegranate rind. The diameters of inhibition zone were 23.5～11.6mm. MIC were 2500～78.1μg/ml and MBC were 5000～156.3μg/ml for Staphylococcus aureus and E.coli. 4) According to the prescriptions of enlarge test was carried out, the mid-experiment drug quality achieved quality standards. The volume variation test and the limit microbe test complied with the general requirements for ointments (appendix1.14). 5) The study of the stability of the ointment was observed by the description, identification, volume variation test and the limit microbe test and assay etc which complied with the general requirements. 6) Neither acute toxic effects nor allergic skin or systematic reactions in intact or injured skin were induced by applications of the oinment. No irritant dermal reaction were observed in intact skin and injured skin. Conclusion: 1) The 5% ointment was uniformity, exquisite, easy to spread on the skin and mucous membrane without thrill. 2) TLC can identified the characteristic spots of the gallic acid in the ointment. According to sample determination result, the pomegranate rind bacteriostatic ointment the content of gallic acid was 0.0667g/g～0.1001g/g. Other requirements must comply with the general requirements for ointments (appendix1.14). 3) Extract, concentrated liquid, freeze-drying power and the ointment have a good antifungal activity in vitro. 4) Through small and medium various technological parameters and quality standard test results showed that the scale-up production was feasible, and the chemical composition and technology transfer rate were high. 5) The quality of the ointment was stability by preserving in 40℃and relative humidity 75% for three months. 6) The oinment was shown to exert no apparent irritant, toxic effects or allergic skin reactions.
Objective: To study the preparation process of pomegranate rind bacteriostatic ointment, antibacterial experiment in vitro, quality standard, stability testing and local application safety test in order to go on further clinic research.Methods: 1) Selected the best prescription ointment matrix with centrifugal test, heat resistance and cold resistance test. 2) The ointment was identified by TLC. The content of the ointment was determined by UV-Vis. 3) The bacteriostasis test in vitro tracked techn ic. 4) The final craft technology was established by mid-experiment. The ointment was determined by relevant rules of ointment according to Chinese pharmacopoeia. 5) Its stability was observed during three months. 6) Local application safety test was observed.It was observed whether the ointment could induce acute toxicity irritation and the allery reaction.Results: 1) To observe the stability of the ointment by cold、hot and centrifuged laboratory experiments. The result showed the ointment was stable. 2) The ointment was identified by TLC. The result showed that the chromatogram of TLC was clear and the resolution was well. After that the quality standard of the ointment was finished. The content of gallic acid in the ointment was determined by UV-Vis, and the average recovery was 99.92% and RSD was 1.02%. 3) Kirby– Bauer method and the tube dilution method were employed for bacteriostasis test in vitro of extract, concentrated liquid, freeze-drying power and ointment of pomegranate rind. The diameters of inhibition zone were 23.5～11.6mm. MIC were 2500～78.1μg/ml and MBC were 5000～156.3μg/ml for Staphylococcus aureus and E.coli. 4) According to the prescriptions of enlarge test was carried out, the mid-experiment drug quality achieved quality standards. The volume variation test and the limit microbe test complied with the general requirements for ointments (appendix1.14). 5) The study of the stability of the ointment was observed by the description, identification, volume variation test and the limit microbe test and assay etc which complied with the general requirements. 6) Neither acute toxic effects nor allergic skin or systematic reactions in intact or injured skin were induced by applications of the oinment. No irritant dermal reaction were observed in intact skin and injured skin. Conclusion: 1) The 5% ointment was uniformity, exquisite, easy to spread on the skin and mucous membrane without thrill. 2) TLC can identified the characteristic spots of the gallic acid in the ointment. According to sample determination result, the pomegranate rind bacteriostatic ointment the content of gallic acid was 0.0667g/g～0.1001g/g. Other requirements must comply with the general requirements for ointments (appendix1.14). 3) Extract, concentrated liquid, freeze-drying power and the ointment have a good antifungal activity in vitro. 4) Through small and medium various technological parameters and quality standard test results showed that the scale-up production was feasible, and the chemical composition and technology transfer rate were high. 5) The quality of the ointment was stability by preserving in 40℃and relative humidity 75% for three months. 6) The oinment was shown to exert no apparent irritant, toxic effects or allergic skin reactions.
引文
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