A Colorimetric Test for the Evaluation of the Insecticide Content of LLINs Used on Bioko Island, Equatorial Guinea

Background: Insecticide-treated nets and indoor residual spraying of insecticides are used as the vector control interventions in the fight against malaria. Measuring the actual amount of deposits of insecticides on bed nets and walls is essential for evaluating the quality and effectiveness of the intervention. A colorimetric "Test Kit" designed for use as a screening tool, able to detect the type II pyrethroids on fabrics and sprayed walls, was used for the first time to detect deltamethrin on long-lasting insecticidal nets (LLINs) deployed on Bioko Island, Equatorial Guinea.

Methods: LLINs were analysed using the colorimetric Test Kit performed in situ, which leads to the formation of an orange-red solution whose depth of colour indicates the amount of type II pyrethroid on the net. The kit results were validated by measuring the amount of extracted insecticide using high-performance liquid chromatography (HPLC) with diode array detection (DAD).

Results: Deltamethrin concentration was determined for 130 LLINs by HPLC-DAD. The deltamethrin concentration of these nets exhibited a significant decrease with the age of the net from 65 mg/m (< 12 months of use) to 31 mg/m (> 48 months; p < 0.001). Overall, 18% of the nets being used in households had < 15 mg/m of deltamethrin, thus falling into the "Fail" category as assessed by the colorimetric Test Kit. This was supported by determining the bio-efficacy of the nets using the WHO recommended cone bioassays. The Test Kit was field evaluated in situ and found to be rapid, accurate, and easy to use by people without laboratory training. The Test Kit was shown to have a reliable linear relationship between the depth of colour produced and deltamethrin concentration (R = 0.9135).

Conclusion: This study shows that this colorimetric test was a reliable method to assess the insecticidal content of LLINs under operational conditions. The Test Kit provides immediate results and offers a rapid, inexpensive, field-friendly alternative to the complicated and costly methods such as HPLC and WHO cone bioassays which also need specialist staff. Thus, enabling National Malaria Control Programmes to gain access to effective and affordable monitoring tools for use in situ.