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EN 1499

Determine Effectiveness of Handwash formulations

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BS EN 1499 : 2013 – Chemical Disinfectant & Antiseptics. Hygienic Handwash (phase 2, step 2 test)

EN 1499 specifies a test method that stimulates practical conditions to determine the efficacy of hygienic hand wash products in reducing transient microbial flora of the hands. To perform EN 1499 test, the hands of volunteers are artificially contaminated with test organisms.

EN 1499 standard is applicable for antimicrobial handwash intended for use in areas and/or situations where disinfection is medically indicated such as –

  • In health care setting such as  hospitals, in community medical facilities, and in dental institutions
  • In clinics of schools, of kindergartens, and of nursing homes 
  • Other areas like laundry and catering service providing direct product supply for patients
  • May also occur in workplace and in home

EN 1499 Test Conditions & Requirement

Mandatory test organisms –  Escherichia coli K12 

Subjects – 12 – 15 volunteer’s hands are artificially contaminated with test organisms. Volunteers for  test are divided into two groups i.e. control and test. For control group,  hands are washed using normal soap (without any antimicrobial components), whereas for the test group, hand washing is done using hand wash product sample (treated one).

Contact times – Between 30-60 second or as per manufacturer’s recommendations

Test Process of EN 1499

EN 1499 Test Method

  • Before contamination, hands of both groups (control and test) are washed to remove dirt and other impurities.
  • Hands are immersed into microbial suspension and then allowed to dry out. Then the fingertips are dipped into tryptic soy broth (TSB) to determine pre-values of viable bacteria present on the hands.
  • Contaminated hands are washed using hand wash product sample. Hand washing time is between 30 – 60 seconds or depends upon manufacturer’s recommendations.
  • At the end of contact time, hands are placed in a neutralizing solution to recover remaining microorganisms.
  • A portion of neutralizing extract is plated and incubated to determine remaining viable microorganisms.
  • Results are calculated by comparing the number of microorganisms obtained from contaminated hands that were washed with product sample to microorganisms recovered from contaminated hands which were washed with normal soap.

Importance of EN 1499 Test

In the health care setting, good hand hygiene practices play a critical role in preventing the direct spread of pathogenic microorganisms. Hygienic hand wash products which come with antimicrobial agents can better protect against harmful microorganisms. EN 1499 test is designed to check the strength of hand wash products intended for use in medical areas.

If you are looking for microbiology labs that do EN 1499 testing for handwash formulations, our experts would be happy to help you. At Microbe Investigations Switzerland (MIS), we offer a comprehensive range of high quality assusrance testing services. We perform an in – depth analysis of product samples in accordance with established international standards.

We also perform EN 1500 testing for hand rub formulations.

To know more about the EN 1499 test method or our testing capabilities, please contact us here.


EN 1499 test is phase 2 step 2 quantitative suspension test. This test stimulates practical conditions to check whether hand wash products are capable of inactivating  transient microbial flora on artificially contaminated hands of volunteers.

EN 1499 test is applicable for hygienic hand wash products designed to be used in areas where disinfection is medically indicated.

EN 1499 test takes 2-3 weeks to complete.

At Microbe Investigations, we test for EN 1499 using the following microbial strains: Escherichia coli (ATCC 8739). Additional stains can be added on customer’s request.

Contact us for more information

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