Infertility affects approximately 186 million people worldwide and 8-12% of couples of reproductive age. Therefore, a comprehensive diagnostic evaluation of infertility is crucial to achieving improvements in targeted prevention and treatment outcomes. The aim of this review is to explore the biochemistry of infertility in order to properly diagnose and treat infertile couples. Recent studies indicate that routine measurement of biochemical parameters reflecting thyroid dysfunction, immunological disorders, autoimmune mechanisms, insulin resistance and malabsorption of selected micro- and macronutrients are required to assess infertility. Due to the complexity of this approach, algorithmic protocols that integrate these biochemical parameters in a dynamic test environment are necessary to provide a more comprehensive diagnostic assessment and more effective treatment strategy for infertile couples.