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Lett Appl Microbiol. 2013 Dec;57(6):517-25. doi: 10.1111/lam.12142. Epub 2013 Aug 27.

Specificity of the neutralizers as the cause of errors in evaluating disinfectant efficacy: an assessment of triclosan.

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Department of Preventive Medicine and Public Health, Faculty of Pharmacy, University of Granada, Campus Universitario de Cartuja, Granada, Spain.


One of the most commonly used methods for assessing disinfectants in vitro is the dilution-neutralization test. Although it is known that using an effective neutralizer is essential for obtaining reliable results, in the case of disinfectants like triclosan, whose neutralization is more complicated, an adequate neutralizer is not always used. Here, we compare the efficacy of different neutralizers described previously, and others that have been modified, against several antiseptic formulations of triclosan. The strains of reference used were Pseudomonas aeruginosa, Enterococcus faecium, Staphylococcus aureus and Escherichia coli. The solutions of antiseptic were triclosan 2% aqueous solution, and three alcoholic solutions containing 2, 0·068 and 0·5% triclosan, respectively. In the dilution-neutralization test, six neutralizers with different degrees of complexity were assayed. Our results show that the antiseptic solutions studied are effective against the strains of reference, with the exception of triclosan 2% aqueous solution against Ps. aeruginosa. Most neutralizers proved ineffective, depending on the antiseptic solution and the bacterial species, particularly in the case of Staph. aureus. These findings confirm a need to test every neutralizer specifically for each disinfectant, at the maximum concentration used, and for every specific bacterial strain.


The choice of an adequate neutralizing agent is essential for obtaining precise results in the evaluation of disinfectants using the dilution-neutralization method. In this study, we show that the neutralization of triclosan and some of its commercial solutions is not a simple task, requiring complex neutralizers. Moreover, the action of the neutralizer is shown to depend on the micro-organism tested. Specificity in view of the disinfectant and the micro-organism calls for performing the neutralizing assay in each particular case of application of the dilution-neutralization method. We also demonstrate that some of the neutralizers described for triclosan in previous studies are not sufficiently effective.


Enterococcus faecium; Escherichia coli; Pseudomonas aeruginosa; Staphylococcus aureus; dilution-neutralization; disinfectants; neutralizers; triclosan

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