One of the global health emergency is represented by the antibiotic resistance, which is considered as a consequence of various patterns of antibiotic consumption. The rise of antibiotics usage has led to an exposure of bacterial communities and ecosystems to a significant amount of antibiotic residues. Therefore, the major future needs should estabilish standardizeds methods for the quantification of the antibiotic residues and antibiotic resistance level which are critical in establishing the link between their presence in the environment and their biological effects. Also, it could be an essential tool in the monitoring and surveillance of antibiotic residues in the environmental matrices as a consequence of inadequate antibiotic use. The analysis of influent and effluent wastewater treatment plant (WWTP) could provide useful information about the medication use and misuse. The aim of the present study is to develop an analytical HPLC/MS-MS method to quantify antibiotics from influents and effluents WWTP. Also, to determine the antibiotic resistance profile of bacteria isolated from wastewaters matrices. This study is focused on the analysis of 7 antibiotics mostly present in the wastewaters of Romania. By incubating the water samples with nutrient agar media, several bacteria were isolated and tested against the detected antibiotics. Results: Using HPLC/MS-MS technique, several antibiotics were detected in the WWTP influent, such as clarithromycin, doxycycline, ciprofloxacin, azithromycin, piperacillin and amoxicillin. After the treatment process, the following antibiotics were detected in effluent samples of the treatment plant clarithromycin, azithromycin, ciprofloxacin. Several bacteria were isolated from the wastewaters and tested against these main antibiotics, all showing resistance to clarithromycin, but also to amoxicillin and norfloxacin. Conclusions: After the wastewater treatment process we detected antibiotic residues in the WWTP effluent, which highlights the ineficiency of wastewater treatment process and a potential risk of waters contamination. Also, rise a concern of negative impact on human and ecosystem health. The next step will be to try and determine the exact type of bacteria isolated and establish the resistance mecanism to antibiotics.