EN 1500 Hygienic Handrub Method

The EN 1500 is a European Standard test method that evaluates the efficacy of a hygienic handrub by measuring the number of viable bacteria remaining on the fingertips after contamination and handrub exposure. A handrub is defined as a treatment that involves rubbing the hands without the addition of water. This method specifically simulates conditions for establishing if a hygienic handrub decreases the release of transient flora from the hands.


In this test, subjects with healthy hands are randomly assigned either a reference control product or the hygienic handrub under evaluation. Following an initial cleansing wash with soft soap to remove natural transient microorganisms, the subject's hands are dried thoroughly with a paper towel.

Typically a prepared pure culture of a non-pathogenic strain of Escherichia coli is used as the inoculum for this test method. Subjects immerse their hands in a volume of the inoculum, up to the mid-carpals, for 5 seconds with their fingers spread apart. The hands are then allowed to air dry for 3 minutes, then the fingertips are sampled by rubbing them into a petri dish with sterile tryptic soy broth (TSB) for pre-values of viable bacteria present on the hands.

Immediately after pre-value sampling the hands are re-inoculated, then subjected to either the reference handrub procedure with propan-2-ol 60% (v/v), or the hygienic handrub being tested per instructions provided by the manufacturer. The fingertips are then sampled again for post-values in the same manner as the pre-values, this time with TSB containing a chemical neutralizer. The samples are then diluted appropriately and plated onto tryptic soy agar (TSA) medium, where they are allowed to incubate 18-24 hours at 36 ± 1ºC, counted, then re-incubated for an additional 24 hours to detect any potential slow growing colonies.

The pre- and post-values recovered from the fingertips are evaluated against one another, resulting in a ratio called the reduction factor. The reduction factor provides a quantitative measure of the antimicrobial efficacy. This is determined by comparing average counts of viable microorganism colony forming units of the pre-values to the average counts after exposure to the hygienic handrub.


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