Consumers use different hair care products to change the physical appearance of their hair, such as shampoos, conditioners, hair dye and hair straighteners. They expect cosmetics products to be available in the market to meet their needs in a broad and effective manner. Evaluating efficacy of hair care products in vitro involves the use of highly accurate equipment. This review aims to discuss in vitro methodologies used to evaluate the effects of hair care products on hair fiber, which can be assessed by various methods, such as Scanning Electron Microscopy, Transmission Electron Microscopy, Atomic Force Microscopy, Optical Coherence Tomography, Infrared Spectroscopy, Raman Spectroscopy, Protein Loss, Electrophoresis, color and brightness, thermal analysis and measuring mechanical resistance to combing and elasticity. The methodology used to test hair fibers must be selected according to the property being evaluated, such as sensory characteristics, determination of brightness, resistance to rupture, elasticity and integrity of hair strain and cortex, among others. If equipment is appropriate and accurate, reproducibility and ease of employment of the analytical methodology will be possible. Normally, the data set must be discussed in order to obtain conclusive answers to the test.