Objectives: To examine the hormonal levels of adolescents with varicocele, specifically focusing on FSH, LH, and inhibin B, treated and followed at Department of Pediatric Surgery at University Hospital of Split. We aim to determine the prevalence of normal, high, or low levels of these hormones in the patient cohort and assess if routine hormonal tests offer any significant diagnostic or prognostic value in the management of varicocele. Materials and methods: This study was designed as a retrospective cohort study conducted at the Department of Pediatric Surgery, University Hospital of Split in Croatia. All data was collected from the hospital’s patient records between 1st of May 2015 to 1st June 2023. This cohort study included a total of 120 participants. The data collection was exclusively extracted from the Department of Pediatric Surgery at the University hospital of Split. Spanning from 2015 to 2021, included exclusively patients treated by a single surgeon. Exclusion criteria for this period involved the absence of recorded LH, FSH, or Inhibin B hormonal levels. For the period 2021 to 2023, the data was collected from the hospital’s system, including patients treated by other surgeons as well. We recorded the patient age, FSH, LH and inhibin B hormonal levels, and whether the patient had undergone surgery. Results: Majority of participants have FSH, LH and Inhibin B levels within the normal reference range. The test statistics shows that there is no significant difference in the levels of LH;(p = 0.081) between operated and non-operated patients. However, there is statistically significant evidence to conclude that the Inhibin B levels;(p = 0.008) were higher for the nonoperated (M=187, SD=77) than operated (M=119, SD=66) patients. There was no significant difference in the FSH values for operated (M=5, SD=4) and non-operated patients (M=3.7, SD=2) conditions; (p = 0.075). Conclusions: In conclusion, this study revealed no statistically significant difference between FSH and LH values in operated and non-operated adolescents. However, a statistically significant difference was found between the inhibin B values in operated and non-operated. However, those who underwent surgery were more likely to have inhibin B values in the lower half of the spectrum compared to their non-operated counterparts. The relevance of monitoring FSH and LH levels does not appear to influence the decision to proceed with varicocelectomy. Further investigations should be done to determine the importance of inhibin B levels in guiding the decision to undergo varicocelectomy.