Background: Recently, there has been an increasing concern in the determination of fructose concentration and enzyme such as glucosidase, present in human seminal plasma. Objective: The main objective of this study is to evaluate the concentration of fructose and the activity of α-glucosidase in human seminal plasma of different infertile categories, normospermia, and control samples and its correlation with semen parameters. Materials and Methods: Infertile semen samples were collected from the azoospermia (n = 15), oligospermia (n = 31), oligoasthenospermia (n = 28), and asthenospermia (n = 32), normospermia (n = 44), and control (n = 13), who attended the semen analysis at Bangalore Assisted Conception Centre Pvt., Ltd, Bengaluru, Karnataka, India. Semen analysis was done for all the samples and categorized by the World Health Organization (2010) protocol. Results: Both fructose and glucosidase were found to be much lower in the case of azoospermia when compared to normospermia and control samples. The concentration was very less 4.247 ± 0.3764 in the case of azoospermia, which is 10 times lesser than the control samples. Fructose concentration and glucosidase activity were compared for all the samples included in this study. Correlation between semen parameters and fructose and glucosidase, respectively, was highly significant with P < 0.0001. Conclusion: Not only with respect to the assessment of sex organ such as seminal vesicle dysfunction but also the role of fructose and glucosidase activity is also significant with other seminal parameters to give a varied dimension in male infertility and male reproductive system. © 2019 JPR Solutions. All rights reserved.