甲基纤维素滴眼液使用和存放过程中的微生物监测分析

Microbial monitoring analysis of methylcellulose eye drops during use and storage

背景 目前大多数滴眼剂为多剂量包装,为保证滴眼液在使用中的安全性,对滴眼液瓶开启后使用过程中的微生物相关研究很有必要. 目的 研究甲基纤维素（MC）滴眼液开启后在社区常温、社区冷藏及病区常温3种环境下使用和存放时微生物污染情况及保持无菌的时间. 方法 将MC滴眼液按照模拟试验的温度、湿度和污染指数等环境的不同分为社区常温组、社区冷藏组和病区常温组,每个环境组取含或不含防腐剂尼泊金乙酯的MC滴眼液各200支,其中分为启封药物和不启封药物.由专业人员开启滴眼液瓶并标注开启时间,模拟点眼操作每次1滴,每日3次,每次药液空气中暴露时间为5～10s,用后分别在相应环境中存放.分别于药瓶开启后1、2、3、4、5、6、7、8、9和10 d的8：00取样,由同一检验员进行细菌培养和真菌培养,并通过全自动微生物分析系统进行菌种鉴定.结果 药瓶开启后社区常温组、社区冷藏组和病区常温组无防腐剂启封药细菌阳性率均在30％左右,而未启封药物未见微生物生长.药瓶开启后10 d,社区常温组、社区冷藏组和病区常温组无防腐剂的启封药细菌培养阳性率分别为30％、32％和36％,而含防腐剂药物的启封药细菌阳性率分别为15％、19％和23％,组内比较差异均有统计学意义（x2=6.452、4.448、4.063,均P＜0.05）;根据Scheffe法计算社区常温组与社区冷藏组、病区常温组与社区冷藏组以及病区常温组与社区常温组间无防腐剂启封药细菌阳性率差值的95％可信区间（CI）分别为-0.166 ～ 0.126、-0.110 ～0.190和-0.088 ～0.208,而含防腐剂启封药物细菌阳性率差值的95％ CI分别为-0.159～0.079、-0.089～0.169和-0.043 ～0.203,尚不能认为不同环境间差异有统计学意义.培养的微生物经鉴定分别为藤黄微球菌、鲁氏不动杆菌、枯草芽孢杆菌、抗辐射不动杆菌、香味菌和木糖葡萄球菌.结论 防腐剂尼泊金乙酯可以降低MC滴眼液使用过程中的微生物污染率;滴眼液的常见使用环境对MC滴眼液的微生物污染情况无明显影响.MC滴眼液污染的菌群均属于大气、土壤等环境菌种,并非眼部常见致病菌。

Background It is imperative for the microbial monitor after opening the bottle of eyedrops in order to ensure the safety during use of ophthalmic solutions with multi-dose packaging.Objective This study was to research the microbiological properties and sterile duration of methylcellulose （MC） eye drops in three common environmental conditions,including room temperature condition of community,refrigeration condition of community and room temperature condition of hospital.Methods MC eye drops were assigned to the community room temperature group,community refrigeration group and hospital room temperature group,and 200 bottles of MC eye drops with or without ethylparaben were collected in each group,including sealed or unsealed drugs at average.The containers of all the eye drops were opened and the opening times were record.The drugs was admistered 1 drop for 3 times per day,with the opening period for 5-10 seconds.Then the drugs were preserved in different environments based on grouping.Microbial isolation and purification were performed by the same lab technician at 8：00 from 1 through 10 days after opening of drugs with automatic microbial analyzer.Results In the unsealed MC eye drops without ethylparaben,the bacterial positive rates were about 30% in the community room temperature group,community refrigeration group and hospital room temperature group,but no microbial colony was seen in the sealed eye drops.Ten days after opening of containers,the bacterial cultured rates were 30%,32% and 36% in the eye drops without ethylparaben in the community room temperature group,community refrigeration group and hospital room temperature group,and those in the eye drops with ethylparaben were 15%,19% and 23%,respectively,showing significant differences between the eye drops with and without ethylparaben （x2 =6.452,4.448,4.063,all at P〈0.05）.The 95% confidence interval （CI） of difference values of intergroup bacterial rates were-0.166-0.126,-0.110-0.190 and-0.088-0.208 between the community room temperature group and the community refrigeration group,between the hospital room temperature group and the community refrigeration group,between the hospital room temperature group and the community room temperature group respectively in the unsealed eye drops without ethylparaben,and those in the unsealed eye drops with ethylparaben were-0.159-0.079,-0.089-0.169 and-0.043-0.203 respectively,indicating insignificant differences among the groups.Cultured bacteria were identified as Micrococcus luteus,Acinetobacter lwoffii,Bacillus subtilis,Acinetobacter radioresistens,Myroides and Staptococcus xylosus.Conclusions Ethylparaben can reduce the contamination rate of microorganisms after opening of MC eye drops.Three environmental conditions do not play an influence on microbial contamination of MC eye drops after opening.The bacteria of contaminated eye drops appear to be common microorganisms in atmosphere and soil,rather than eye common pathogens.