In metallurgical industry, water-soluble cutting fluids are a frequent cause of occupational irritation and/or contact dermatitis in operators. Inhalation of aerosols from cutting fluids can cause irritation to the throat, nose and lungs, while exposure is often associated with chronic inflammation of the bronchi, hypersensitivity pneumonitis and worsening of pre-existing respiratory issues. Gram-negative, Gram-positive bacteria and mycobacteria isolated from cutting fluid has generated great concern due to their ability to act as potential pathogens, which justify why biocides are generally incorporated into the cutting fluid formulation by the manufacturer. Alkanolamines have been used in metallurgical fluids as corrosion inhibitors, but they also have an antimicrobial effect and can be used as an alternative way to contain bacterial proliferation. Therefore, the objective of this work is to evaluate the biological activity of alkanolamines against the bacteria Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), using the determination of the minimum inhibitory concentration (MIC). The standard E. coli and S. aureus strains were reactivated, seeded on agar and the inoculum standardized at 106 CFU/mL. The MIC was determined by the serial microdilution method of different alkanolamine solutions. The results demonstrated that different alkanolamines showed a significant reduction in the microbiological growth of standard Gram-negative and Gram-positive strains, also acting as a bacteriostatic function in industrial fluids.