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Arm-in-Cage Test for Mosquito Repellents
Standard technique for testing the efficacy of mosquito repellents.
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Summary
The Arm-in-Cage test is used to assess effectiveness of mosquito repellent products in preventing mosquito landings and bites on human skin. Our laboratory conducts arm in cage studies to deliver reliable efficacy data that manufacturers can trust for product performance, development, optimization, and market launch.
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Quick understanding about test
Arm-in-Cage Mosquito Repellent test - Standard technique for testing the efficacy of mosquito repellents.
Used for testing topical repellent formulations (lotions, creams, sprays) and repellent-impregnated textiles.
- Aedes,
- Anopheles,
- Culex species are the required mosquito strains for this test.
- Conventional method: A repellent is applied to a volunteer's forearm and exposed to 200 mosquitoes every 30 minutes. This process continues for up to 8 hours or until the repellent stops working.
- Modified methods: 30 mosquitoes are used for the test. Repellent is applied to a volunteer's forearm and tested for efficacy during mosquito exposure. Know more
- Accurate measurement of repellent efficacy and duration of protection.
- Comparison of different formulations.
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What Is the Arm-in-Cage Test?
The arm-in-cage test is an internationally accepted method used to evaluate the efficacy of mosquito repellents intended for human use. It provides required data to demonstrate to assess if a product offers reliable protection against mosquito bites.
Mosquitoes acting as vectors for diseases such as malaria, chikungunya, Zika, and yellow fever and hence effective repellent performance is a public health priority in both European and US markets. Regulatory bodies rely on standardized efficacy data to support product claims and ensure consumer protection.
This test is applicable to topical repellents, including creams, lotions, sprays, as well as repellent-treated textiles
Why Arm-in-Cage Test Is Used for Mosquito Repellents
The arm-in-cage test provides accurate and reliable data on the duration and level of protection provided by different repellent formulations. This aids in the development and optimization of repellent formulations. The test mimics real-world conditions thus making it an essential test for manufacturers to ensure their products meet regulatory standards.
Mosquito Species Used in Arm-in-Cage Tests
At MIS, we perform the arm-in-cage test for mosquito repellent using the following mosquito strains:
- Aedes
- Anopheles
- Culex species
Products Applicable for Arm-in-Cage Testing
Arm-in-cage test is used to determine the efficacy of topical repellents (cremes, lotions, and spray formulations) and impregnated textiles.
Arm-in-Cage Test Methodology
Conventional Arm-in-Cage Test
- The test is conducted using 200 host-seeking female mosquitoes, held in the test cage.
- The repellent formulation is applied to the volunteerโs forearm and exposed to mosquitoes every 30 minutes.
- This exercise is repeated for 8 hours after applying the repellent or until it becomes ineffective.
Modified Arm-in-Cage Tests (Customized)
Conventional arm-in-cage test procedure has been modified according to the guidelines published by the American Environmental Protection Agency (EPA) providing a few significant benefits over conventional arm-in-cage methods.
Modified Arm-in-Cage Test Procedure
- The test is conducted using 30 host-seeking female mosquitoes which are held in the test cage.
- Tropical repellent formulations are applied to the volunteerโs forearm and exposed to mosquitoes every 30 minutes.
- For Textiles, the fabric is wrapped on the forearm of 4 volunteers (2 males and 2 females).
- This exercise is repeated for 8 hours after applying the repellent or until it becomes ineffective.
A comparison between the test results of the modified arm-in-cage test and conventional arm-in-cage test using a standard repellent revealed that the protection times obtained in the modified setup were better.
Comparison between Conventional Arm in cage vs Modified Arm in cage
Parameter | Conventional Arm-in-Cage Method | Modified Arm-in-Cage Method |
Number of mosquitoes used | High, typically 200 female mosquitoes | Low, typically ~30 female mosquitoes |
Air ventilation | No dedicated air ventilation system | Dedicated Air ventilation system to prevent accumulation of odors and materials |
Cage contamination risk | Higher risk of contamination due to accumulation of residual test material | Reduced chance of contamination due to controlled exposure and ventilation |
Forearm exposure | Treated forearm is directly exposed to mosquitoes inside the cage | Forearm exposed through a fixed opening or window |
Passing Criteria - Complete Protection Time (CPT) Explained
CPT is determined by measuring the duration between repellent application and the first confirmed bite. This is defined based on either the occurrence of two or more bites on the treated arm or a bite followed by another within 30 minutes (according to ECHA (European Chemicals Agency) Transitional Guidance). The longer the CPT time, the greater the effectiveness of repellents.
Arm-in-cage test vs Other Tests
Parameter | Room Test | Arm-in-Cage Test | WHO Cone Test |
Purpose | Evaluates mosquito repellency / knockdown / mortality in an enclosed room | Measures mosquito (bite inhibition / repellency) on human skin | Assesses residual insecticidal efficacy of treated surfaces |
Mandatory insects | Aedes aegypti, Culex quinquefasciatus, Anopheles stephensi (depending on claim) | Anopheles, Aedes, or Culex mosquitoes | Anopheles, Aedes, or Culex mosquitoes (as per protocol) |
Scope of products | Coils, vaporizers, mats, aerosols, spatial repellents, electric devices | Topical repellents (lotions, creams, sprays, gels) | Wall paints, treated nets, sprays, residual surface treatments |
Passing criteria | With volunteers – Complete Protection Time (CPT) is Without volunteers – โฅ80% knockdown and โฅ90% mortality after 24 hours | Higher Complete Protection Time (CPT) means higher the protection | The product must demonstrate 100% mortality after 24 hours |
Regulatory Requirements for Conducting an Arm-in-Cage Test
Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) Informed Consent
- Subjects must be fully informed about the test’s nature, purposes, and foreseeable health consequences.
EPAโs Human Subjects Rule
- Tests must adhere to EPA regulations for the protection of human subjects set out in 40 CFR part 26, subparts K, L, and M. These regulations apply to any research involving intentional exposure of human subjects that is intended for submission to the EPA under pesticide laws.
Good Laboratory Practice (GLP) Standards
- Research must follow GLP standards
State Requirements
- Ensure compliance with relevant state laws and regulations
Documentation and Record-keeping:
- Maintain detailed records, including proposals, Institutional Review Board (IRB) reviews, consent documents, progress reports, and deviations.
Ethical Considerations:
- Minimize risks to subjects, ensure equitable selection, and properly document informed consent.
- Justify research ethically, consider alternatives, and include risk minimization strategies in the protocol.
Request Arm-in-Cage Testing
Are you in the process of developing or optimizing mosquito-repellent products? Microbe Investigations Switzerland (MIS) offers Arm-in-Cage testing to rigorously evaluate the effectiveness of mosquito repellents. Our state-of-the-art lab is equipped to conduct comprehensive efficacy and safety tests, ensuring precise and reliable results. Trust MIS to support your product development with our comprehensive testing services.
Get a quote on Arm in cage testing
Contact our experts today to validate your mosquito repellentโs efficacy with a standardized Arm-in-Cage test.
Frequently Asked Questions
DR. Martinoz Scholtz
The arm-in-cage test measures the efficacy of a topical repellent to prevent mosquito landings and/or bites on treated human skin under controlled laboratory conditions, providing a quantitative assessment of repellency over time.
Complete Protection Time (CPT) is defined as the time interval between repellent application and the first confirmed mosquito landing with probing or biting during scheduled exposures.
Arm-in-cage repellent testing is conducted according to recognised guidelines such as WHO (WHOPES), US EPA OPPTS 810.3700, ASTM E951, ECHA BPR Efficacy Guidance, and Health Canada (PMRA).
Yes, this test is used for topical repellents (lotions, creams, sprays) and for repellent-impregnated textiles.
At MIS, we perform the arm-in-cage test for mosquito repellent using the following mosquito strains: Aedes, Anopheles, and Culex species.
After application, exposures are typically repeated every 30 minutes for 8 hours or until protection fails, allowing CPT determination.
Key influencing factors include:
- Mosquito age, hunger, and density
- Ambient temperature and humidity
- Volunteer attractiveness and skin chemistry
The arm-in-cage test is a laboratory method that measures repellent effectiveness under controlled conditions. While it provides reliable comparative data, it does not fully reflect real-world use, so field testing is recommended for a complete assessment.
Ethical safeguards include informed consent, use of disease-free laboratory mosquitoes,limiting exposure time, medical oversight and bite management
The modified arm-in-cage setup uses a smaller cage and only 30 mosquitoes instead of 200, with an integrated air ventilation system that prevents odor buildup and reduces contamination risk.
Yes, arm-in-cage results vary by mosquito species and density. . Species aggressiveness and mosquito density significantly influence landing and biting rates, affecting CPT outcomes.
Arm-in-cage tests are designed to measure repellent effectiveness and are not used for assessing attractant properties.
Limitations include
- It doesnโt completely imitate real world conditions.
- It doesnโt give results of long-term wear effects.
The effectiveness is tested by applying the repellent to a volunteer’s forearm and exposing it to mosquitoes in a cage. The test measures the Complete Protection Time (CPT), which is the duration from repellent application until the first confirmed bite.
Mosquito repellents work by masking human odors or emitting substances that mosquitoes find unpleasant, thus preventing them from landing and biting.
Mosquitoes are selected based on the species relevant to the geographic area and the target market. Species like Aedes, Anopheles, and Culex are typically used.
Yes, the results from these tests can be used for regulatory approval, provided that standard guidelines from organizations like the WHO or EPA are followed during testing.
The repellent should be spread evenly on the volunteerโs forearm. The amount and application method should follow the specified guidelines in the test protocol and manufacturerโs instructions.
Generally, five to ten volunteers are involved in arm-in-cage tests for product registration, while initial product development might use fewer participants.
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