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The current study was applied on 200 cows milk samples, which collected randomly from different cities in Libyan, were included 53 cow raw milk, 27 pasteurized milk and 120 sterilized milk. Examined qualitative by using Delvo test SP NT, the results revealed that antibiotic residues were 75.47% and 48.15% in cow raw milk and pasteurized milk, respectively. And all the sterilized milk tested were 100% negative for a test. High liquid performance chromatography (HPLC) were used as confirmatory method for identification of Beta-lactam residues in 14 raw milk positive with Delvo test SP NT. The detected of Ampicillin, Amoxicillin, Tetracycline and Oxytetracycline residues in cow raw milk were 35.71%, 71.42%, 85.71% and 78.57%, respectively. Concentration limits a mean value 4.88±1.02µg/kg, 12.22±6.18µg/kg, 86.24±56.60µg/kg and 108.40±54.15µg/kg for Ampicillin, Amoxicillin, Tetracycline and Oxytetracycline residues, respectively. In raw milk. Confirmed, Amoxicillin, Tetracycline and oxytetracycline residues were 2/6 (33.33%), 5/6 (83.33%) 5/6 (83.33%) and 6/6 (100%) In pasteurized milk, respectively. With concentration limits the mean value 3.45±0.25µg/ kg Ampicillin, 14.06±3.97µg/kg 112.50±89.04µg/kg and 124.30±81.74µg/kg, for Ampicillin, Amoxicillin, Tetracycline and Oxytetracycline residues in pasteurized milk, respectively. In this study examined 20 sterilized milk, showed that antibiotic residues were not detected by used high-performance liquid chromatography.