The present study aimed to validate a control method on the gas chromatography system (GC) based on the experimental design strategy, to examine the changes and correlation between the fermentation process and the quality of alcoholic and non-alcoholic beer product, especially the formation of diacetyl. On the other hand, spectrophotometric methods were applied to the determination of polyphenols content and the potential antioxidant activity of beer during different fermentation processes. with this aim, three modes of barley fermentation were used, specifically classical fermentation, stopped fermentation and thermal process. The results showed that the different fermentation modes had a major impact on diacetyl production. The highest concentration was obtained using stopped fermentation 0.36 mg/L, the lowest concentration value 0.07 mg/L was detected using the thermal process. Monitoring the increase of oxygen concentration between fermentation, filtration, and filling of the final product (32, 107, 130 ppm, respectively) has a significant impact on the concentration of diacetyl. The obtained results of spectrophotometric analysis showed that the total antioxidant activity changed during beer fermentation process and demonstrate that the extend of the antioxidant activity was very much dependent on the total polyphenolic content with a higher value in Hopped wort (13.41%, 65 mg GAE 100 mL−1, 28 mg CE 100 mL−1) for antioxidant potential, total phenolic content, and total flavonoids content, respectively, whereas the lowest values was detected in Non-alcoholic beer using thermal process (7.24%, 35 mg GAE 100 mL−1, 10 mg CE 100 mL−1) for antioxidant potential, total phenolic contents, and total flavonoids contents, respectively. Based on the results achieved, we reveal the impact of the fermentation process on the nutritional value of the final product.