The viral infection known as hepatitis C (HCV) is a significant threat to health in Pakistan. Although powerful antiviral therapies are readily available, the disease load in the general population as a whole continues to rise. This may be because the infection is non-symptomatic, and delaying diagnosis until the disease has progressed significantly. Objectives: The study aims to contrast the biochemical profiles of patients with HCV before and after treatment with Sofosbuvir. Material and Methods: This was a cohort type of study with a sample size of 106. Patients with hepatitis C who went to the Medical Unit at the District Head Quarters Hospital in Rawalpindi between May 2019 and January 2020 were enrolled. To learn more about the patients' personal and medical histories, a questionnaire was created. Venipuncture was used to draw blood, and the Beckman Coulter AU480 chemistry analyzer was used to calculate serum uric acid levels, total cholesterol levels, triglycerides levels, ALT levels, and TSH levels. Performa was used to log data, which was then transferred to SPSS for analysis. Results: In total 106 patients were studied, 75% were from Rawalpindi and 25% were from other areas of Pakistan. The mean age of patients enrolled in this study was 43.5 ± 12.3 years, 66.04% of the HCV patients have a negative family history, and the highest percentage (94.34%) of the patients are married. Patients were divided into two groups based on the duration of anti-HCV therapy Sofosbuvir as Pre- therapy and Post therapy. The Pre and Post Sofosbuvir therapy found non-significant results of uric acid levels Mean ± S.D of (4.79 ± 1.57) v/s (5.22 ± 1.51) with p-value (>0.05), total cholesterol levels Mean ± S.D of (164.2 ± 37.22) v/s (174.1 ± 43.27) with p-value (>0.05), triglycerides levels Mean ± S.D of (136.7 ± 37.9) v/s (110 ± 20.3) with p-value (>0.05) and TSH levels Mean ± SD (2.10 ± 1.97) v/s (2.93 ± 1.82) with p-value (>0.05). But the significant result was found for ALT levels (90.3 ± 104.1) v/s (43.6 ± 33.8) with p-value (<0.05). Conclusion: This study's findings support the use of Sofosbuvir in the treatment of chronic HCV due to its low risk and impressive effectiveness. Related to decreased alanine transaminase (ALT) levels and the suppression of viral replication.