The paper presents a method for determining the end of service life of all-rubber and polymer-coated gloves protecting against mineral oils and selected mechanical factors. Analysis of variance was conducted to test for differences between the protective parameters (abrasion resistance, cutting resistance, tear resistance, mineral oil permeation resistance) and an ergonomic parameter (bending rigidity) determined for five models of protective gloves following a simulated use procedure. The results show that both protective and ergonomic parameters are affected to the greatest degree by the period of use under conditions of exposure to mineral oil: 2 h, 4 h, and 8 h for polymer-coated gloves and 4 h, 8 h, and 16 h for all-rubber gloves. It has been found that gloves lose their protective properties over time under simulated conditions of use and their performance levels may no longer comply with the relevant standards. Based on the tests conducted, a test protocol has been developed for end-of-service-life evaluation of gloves protecting against mechanical factors and mineral oils.The current system of harmonized standards for testing protective gloves provides information about protection levels only for new products. The proposed protocol for end-of-service-life evaluation of protective gloves exposed to mineral oils and mechanical factors may serve as a useful tool for employers and workers by providing reliable information about the approximate actual period over which gloves may be used safely in the workplace.