This study aimed to investigate the residue levels of Tetracycline (TC), Oxytetracycline (OTC), Doxycycline (DOC), and Chlortetracycline (CTC), in 40 samples of poultry meat (n = 40) and feed (n = 10) samples along with the biochemical properties of poultry chicken blood by using a confirmatory multi-class LC–MS/MS method. All the samples are collected from the major two cities (Dhaka, Rajshahi) For efficiency of the study, the total experiment is divided into two major groups (phase1: Blood and feed property analysis; Phase- 2: Antibiotic residues analysis). From the first experiment in poultry chicken blood sample analysis, it was found that the, cholesterol, uric acid, phosphorous, HDL, glucose and total protein were 107.15, 5.664,5.101 ,19.545(mg/dl), 7.238(mol/L), and 4.442(g/dl), in Dhaka and 87.191, 5.75, 4.669, 21.82(mg/dl) , 5.637(mol/L), and 5.565(g/dl) in Rajshahi city according to blood sample analysis.