盐酸奥洛他定滴眼液中抑菌剂的评价

目的：考察不同厂家不同批次盐酸奥洛他定滴眼液中抑菌剂苯扎氯铵的抑菌效力及细胞毒性，探索既满足抑菌效力又保证细胞毒性较低的抑菌剂最低浓度。方法：采用《中华人民共和国药典》（简称《中国药典》）2015年版四部附录〈1121抑菌效力检查法〉进行抑菌效力试验，以无抑菌剂的盐酸奥洛他定滴眼液为对照，考察含不同浓度抑菌剂滴眼液的抑菌效力情况。同时以HConEpic人结膜上皮细胞为研究对象，考察不同浓度的苯扎氯铵溶液，含不同浓度苯扎氯铵的盐酸奥洛他定滴眼液，以及不同厂家不同批号的盐酸奥洛他定滴眼液的细胞毒性。结果：11批样品均符合《中国药典》2015版四部〈1121抑菌效力检查法〉要求，通过不同浓度抑菌剂的考察，证明含0.005%苯扎氯铵的盐酸奥洛他定滴眼液的抑菌效力结果即可满足药典要求。盐酸奥洛他定无明显的细胞毒性，添加不同浓度的苯扎氯铵后，细胞毒性与浓度呈明显的正相关（0.01% > 0.005% > 0.003% > 0.001%）。结论：含0.005%苯扎氯铵的盐酸奥洛他定滴眼液抑菌效力即可满足药典要求，而含0.005%苯扎氯铵的细胞毒性比含0.01%的细胞毒性降低较多，因此，有必要降低盐酸奥洛他定滴眼液中苯扎氯铵浓度。

Objective: To investigate the antibacterial activity and cytotoxicity of the bacteriostatic benzalkonium chloride in different batches of olopatadine hydrochloride eye drops from different manufacturers, and to explore the lowest concentration of bacteriostatic agent that meets both bacteriostatic efficacy and low cytotoxicity.Methods: The antibacterial efficacy test was carried out according to the 1121 bacteriostatic efficacy test in the Pharmacopoeia of the People's Repulic of China 2015 edition (ChP 2015).The olopatadine hydrochloride eye drops without bacteriostatic agent were used for the control.And the antibacterial efficacy of the eye drops containing different concentrations of bacteriostatic agents were investigated.At the same time, HConEpic human conjunctivalepithelial cells were used as research objects to investigate the different concentrations of benzalkonium chloride in olopatadine hydrochloride eye drops, with different batches or different manufacturers.Results: All samples were in compliance with the requirements of the 1121 bacteriostatic efficacy test in ChP 2015.The antibacterial efficacy of olopatadine hydrochloride eye drops containing 0.005% benzalkonium chloride can meet the requirements of ChP 2015.There was no obvious cytotoxicity of olopatadine hydrochloride, and there was a significant positive correlation between cytotoxicity and concentration of benzalkonium chloride (0.01% > 0.005% > 0.003% > 0.001%).Conclusion: The antibacterial efficacy of olopatadine hydrochloride eye drops containing 0.005% benzalkonium chloride can meet ChP requirements, while the cytotoxicity of 0.01% benzalkonium chloride is much more than that of 0.005%.It is both possible and necessary to reduce the concentration of benzalkonium chloride in olopatadine hydrochloride eye drops.

[1] 李宁,陈松杰.聚季铵盐-1和苯扎氯铵在眼用制剂中的应用研究进展[J].今日药学,201525(2):140 LI N,CHEN SJ.Progress in the application of polyquaternary ammonium salt-1 and benzalkonium chloride in ophthalmic preparations[J].Pharm Today,201525(2):140
[2] FREEMAN PD,KAHOOK MY.Preservatives in topical ophthalmic medications:historcal and clinical perspectives[J].Expert Rev Ophthalmol,2009,4(1):59
[3] TU EY.Balancing antimicrobial efficacy and toxicity of currently available topical ophthalmic preservatives[J].Saudi J Ophthalmol,2014,28(3):182
[4] LIANG H,PAULY A,RIANCHO L,et al.Toxicological evaluation of preservative-containing and preservative-free topical prostaglandin analogues on a three-dimensional-reconstituted corneal epithelium system[J].Br J Ophthalmol,2011,95(6):869.
[5] DEBBASCH C,BRIGNOLE F,PISELLA PJ,et al.Quaternary ammoniums and other preservatives'contribution in oxidative stress and apoptosis on Chang conjunctival cells[J]Invest Ophthalmol Vis Sci,2001,42(3):642
[6] 张琛,王志昕,邓世靖,等.防腐剂对兔角膜上皮细胞毒性及玻璃酸钠保护作用的试验研究[J].眼科新进展,2008,28(2):85 ZHANG C,WANG ZJ,DENG SJ,et al.Cellular toxicity of preservatives in corneal and conjunctival epithelial cells and cytoprotective action of sodium hyaluronate[J].Rec Ads Ophthalmol,2008,28(2):85
[7] 李珏,王知坚,项露婷,等.苄达赖氨酸滴眼液中抑菌剂的检测与评价[J].药物分析杂志,2015,35(5):884 LI J,WANG ZJ,XIANG LT,et al.Evaluation and testing of antimicrobial preservatives in bendazac lysine eye drops[J].Chin J Pharm Anal,2015,35(5):884
[8] 肖璜,林吉恒,陈万胜,等.氯霉素滴眼液处方中尼铂金乙酯含量合理性探讨[J].中国药师,2014,17(5):785 XIAO H,LIN JH,CHEN WS,et al.Expolration of rational ethylparaben contents in chloramphenicol eye drop[J].China Pharm,2014,17(5):785
[9] 林吉恒,肖璜,陈万胜,等.氯霉素滴眼液处方中硫柳汞含量的合理性分析[J].中国药师,2014,17(10):1687 LIN JH,XIAO H,CHEN WS,et al.Analysis of rational thiomersal contents in chloramphenicol eye drop[J].China Pharm,2014,17(10):1687
[10] 中华人民共和国药典2015版.四部[S]:2015:151 ChP 2015.Vol Ⅳ[S].2015:151
[11] 肖璜,杨美琴,王似锦,等.国产滴眼液中常用抑菌剂的应用分析与评价[J].药学研究.2017,36(9):519 XIAO H,YANG MQ,WANG SJ,et al.Analysis and evaluation of bacteriostatic agent appliacation in domestic eye drops[J].J Pharm Res,2017,36(9):519
[12] 朱世真,王丽芳,王尊文,等.常用滴眼剂抑菌效力的考察[J].中国药品标准,2016,17(2):95 ZHU SZ,WANG LF,WANG ZW et al.Investigation on bacteriostatic efficacy of common eye drops[J].Drug Stand China,2016,17(2):95
[13] PELLINEN P,HUHTALA A,TOLONEN A,et al.The eytotoxic effects of preserved and preservative-free prostaglandin analogs on human coreal and conjunctival epithelium in vitro and the distribution of benzalkonium chloride homologs in ocular surface tissues in vivo[J].Curr Eye Res,2012,37(2):145
[14] 张超,赛景影,张晓天,等.一次性氧氟沙星滴眼液抑菌效力的检测及其结果评价[J].吉林大学学报(医学版),2014,40(1):113 ZHANG C,SAI JY,ZHANG XT,et al.Detection of bacteriostatic effect and evaluation of disposable ofloxactin eye drops[J].J Jilin Univ(Med Ed),2014,40(1):113
[15] 罗庆进,陈吉祥,李素琴,等.一种不含抑菌剂的氧氟沙星滴眼液及其制备工艺:中国,CN105232452A[P].2015-10 LUO QJ,CHEN JX,LI SQ,et al.An Ofloxacin Eye Drop without Antimicrobial Agent and Its Preparation Technology:China,CN105232452A[P].2015-10