Chloramphenicol (CAP) is a broad-spectrum antibiotic widely applied in veterinary practices. Continual use of CAP in livestock production may lead to antibiotic resistance and health-related hazards. In this study, a rapid and highly sensitive analytical method based on ultra-performance liquid chromatographytandem mass spectrometry (UPLC–MS/MS) was developed and validated to detect and quantify there is due of CAP in poultry meat regarding the safety of humans and animals. Total 80 samples of poultry meat were collected from different poultry farms, and poultry meat sellers in three upazilas namely- Sadar, Mirzapur and Ghatail of Tangail district in the Dhaka division. The test analysis was performed using matrix-matched liquid chromatography-mass spectrophotometry. The chromatographic separation of the CA Presidue was carried out at 40°C temperature on a reverse-phase C18 column using a binary gradient pump, and quantification was performed by LC-MS/MS in electrospray mode. Mobile phase constituents were solvent (a) deionized water, and (b) acetonitrile. The flow rate was 0.35 mL/min and the total run time was 5 min. The method was validated in terms of selectivity, linearity, recovery, and precision following the 2021/808/EC guidelines and acceptance criteria were met in all the cases. The relative standard deviation (RSD) for precision was <11%. The linearity of the calibration curves was excellent (R2 >0.999) at concentrations of 0.25, 0.50, 0.75, 1.0, 2.0, and 5.0 μg/kg for matrix-matched CAP standard, and the range of linearity of this method was 0.0-5.0 μg/kg with R2 value greater than 0.99. The decision limit (CC-a), and detection capability (CCβ) were 0.29 μg/ kg, and 0.31 μg/kg respectively, and the recovery percentages ranged from 94 to 100 %. In this study, the levels of CAP residue in tested poultry meat samples were found below the detection limit. The overall parameters of the proposed method met the validation criteria, and the method proved to be suitable for CAP residues determination in poultry meat samples. Thus, this method could be a precise and highly desirable analytical procedure for rapid and routine analysis of CA Presidues in poultry meat, and obtained tested results in this study could be an authentic data to ensure the chloramphenicol free safety poultry meat for human consumption.

Quantification of vitamins in complex matrices such as feed additives is a time-consuming analytical procedure. In this study, a simple and precise in-house High Performance Liquid Chromatography (HPLC) method was developed and validated for the simultaneous detection and quantification of four fat-soluble vitamins such as vitamin A, D3 , E, and K3 in feed additives. The HPLC method was developed and validated using reversed-phase column chromatography. The chromatographic separation of the vitamins was carried out at 25° C temperature on a reverse-phase C18 column using a binary gradient pump mode. Mobile phase constituents were solvent (a): deionized water and (b) methanol. Detection was performed with HPLC ultraviolet/visible detection set at 325, 265, 230, and 254 nm wavelength for vitamin A, D3 , E, and K3 respectively. The flow rate was 1.0mL/min and the total run time was 20min. The method was validated according to the guidelines of the International Conference on Harmonization (ICH) and Food and Drug Administration (FDA), USA, and acceptance criteria for system suitability, specificity, linearity, accuracy, and precision were met in all the cases. The Relative Standard Deviation (RSD) for system suitability and precision was <2% for all the studied vitamins. The linearity of the calibration curves was excellent (R2 >0.999) at concentrations of 2.5, 5.0, 7.5, 10.0, 15.0, and 20.0 µg/mL for all vitamins, and the range of linearity of this method was 0.0-50.0 μg/mL with R2 value greater than 0.999. The limits of detection values were 0.0022, 0.0012, 0.0022, and 0.0020 µg/ mL for vitamin A, D3 , E, and K3 , respectively, and the limits of quantification values were 0.0066, 0.0038, 0.0066, and 0.0061 µg/mL for vitamin- A, D3 , E, and K3 respectively. The recovery percentages ranged from 85% to 103%, and the robustness of the method is also high with excellent reproducibility. The overall parameters of the proposed method met the validation criteria and this method could be a precise and highly desirable analytical procedure for accurate quantification of four fat-soluble vitamins such as A, D3 , E, and K3 in feed additives using a single chromatographic run.