目的建立硝酸咪康唑、酮康唑、曲安奈德益康唑3種咪唑類抗真菌乳膏劑的微生物限度檢查方法。方法采用含5%聚山梨酯80的pH 7.0無菌氯化鈉蛋白胨緩沖液制備3種乳膏劑1：10供試液，并稀釋至1：20和1：50，分別加入金黃色葡萄球菌、銅綠假單胞菌、枯草芽孢桿菌、白色念珠菌、黑曲霉接種無菌平皿中，按照《中國(guó)藥典》2015年版四部1105考察平皿法及稀釋-平皿法計(jì)數(shù)方法適用性；分別取3種乳膏劑加入無菌pH 7.0無菌氯化鈉蛋白胨緩沖液和十四烷酸異丙酯兩相系統(tǒng)中萃取，取萃取后的下層水相的上清液薄膜過濾處理后，分別加入5種試驗(yàn)菌，進(jìn)行萃取-薄膜過濾法計(jì)數(shù)法適用性試驗(yàn)；分別取未經(jīng)萃取-薄膜過濾處理和經(jīng)過萃取-薄膜過濾的1：10供試液，接種至不同體積的胰酪大豆胨液體培養(yǎng)基中，按照《中國(guó)藥典》2015年版四部1106進(jìn)行控制菌檢查適用性試驗(yàn)。結(jié)果通過平皿法或稀釋處理，3種乳膏劑的3種細(xì)菌類試驗(yàn)菌回收率即符合要求；3種乳膏劑對(duì)白色念珠菌、黑曲霉均有較強(qiáng)的抑菌作用，無論是平皿法還是稀釋處理后的平皿法，2種試驗(yàn)菌回收率均遠(yuǎn)小于0.5，不符合方法適用性要求。萃取-薄膜過濾法可有效去除3種咪唑類抗真菌乳膏劑對(duì)各試驗(yàn)菌的抑菌作用，3種乳膏劑的試驗(yàn)菌回收率均在0.5~2.0，符合計(jì)數(shù)方法適用性要求。經(jīng)過萃取-薄膜過濾處理的供試液控制菌檢出優(yōu)于未經(jīng)處理的供試液。結(jié)論采用萃取-薄膜過濾法可解決樣品抑菌性的去除和濾膜堵塞的問題，適用于3種咪唑類抗真菌類乳膏劑的微生物限度檢查。

Objective To establish the microbial limit test method for three kinds of imidazole antifungal creams, such as miconazole nitrate, ketoconazole, and triamcinolone econazole. Methods Sterile sodium chloride peptone buffer of pH 7.0 containing 5% polysorbate 80 was used to prepare three kinds of cream 1∶10 for test solution, and dilute to 1∶20 and 1∶50, respectively. Test solution was added with Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans, Aspergillus niger, and inoculated sterile plate. According to the 1105 of Volume four from the "Chinese Pharmacopoeia" 2015 edition, the applicability of plate method and dilution plate method was investigate. Cream was added to a two-phase system of sterile pH 7.0 sterile sodium chloride peptone buffer and isopropyl myristate. Add five kinds of test bacteria and perform the counting method suitability test. The test solution of 1∶10 with and without extraction membrane filtration treatment were inoculated into different volumes of tryptic soy peptone liquid culture medium, according to 1106 of Volume four from the "Chinese Pharmacopoeia" 2015 edition to carry out control bacteria test suitability test. Results Through the plate method or dilution treatment, the recovery rates of three kinds of bacteria in the three creams met the requirements; the three creams had strong antibacterial effect on Candida albicans and Aspergillus niger, and the recovery rates of the two kinds of bacteria were far less than 0.5, which did not meet the applicability requirements of the method. The recovery rate of each test bacteria for the determination of total aerobic microbial count, total combined yeasts and molds count all met the requirements of 0.5 ~ 2.0, and the detection of control bacteria in the test solution after extraction-membrane filtration treatment was better than that of the untreated Test solution. Conclusion The extraction-membrane filtration method can solve the problem of sample antibacterial removal and filter clogging. It is suitable for the microbial limit inspection of three imidazole antifungal creams.

R965.3

陜西省重點(diǎn)研發(fā)項(xiàng)目（2019SF062）