Field workers in Canada are occupationally exposed to pesticides under diverse environmental conditions of temperature, solar radiation (e.g. ultra violet (UV)), air humidity, etc. that may affect the barrier integrity of chemical protective clothing (CPC). The present study reports a new flow-cell design for testing pesticide permeation in CPC under precisely controlled environmental conditions. Pesticide analysis was conducted by liquid scintillation counting of ¹⁴C-ring-labelled 2,4-dichlorophenoxyacetic acid (2,4-D). Studies conducted to determine the permeation of 2,4-D under light (UV) and dark conditions in black neoprene, natural rubber (NR), nitrile butyl rubber, polyvinyl chloride (PVC), and Viton^R gloves demonstrated low permeation overall with a maximum of 3.7 ± 1.48% permeation by 48 h in PVC gloves. Significantly greater (P < 0.05) permeation of 2,4-D was observed in NR gloves exposed to UV for 24 h. Advantages of our in-house developed Automated In-Vitro Diffusion Analysis (AIDA) procedure to the standard American Society for Testing and Materials (ASTM) F739-85 method will be discussed.