Gentamicin, an aminoglycoside antibiotic, plays a vital role in treating serious Gram-negative infections. Due to its narrow therapeutic window and associated risks of nephrotoxicity and ototoxicity, precise therapeutic drug monitoring is essential to ensure patient safety. Various analytical methods have been developed for quantifying gentamicin in pharmaceutical formulations and biological fluids. Traditional microbiological and spectrophotometric methods, while cost-effective, often lack specificity. More advanced chromatographic techniques, such as HPLC-UV and HPLC-MS/MS, provide high sensitivity and precision, whereas immunoassays like ELISA and radioimmunoassay offer rapid detection. Emerging innovative approaches, including capillary electrophoresis and chemiluminescence, present environmentally friendly alternatives. Among these, HPLC-MS/MS stands out as the most precise and reliable technique due to its capability to detect individual gentamicin components within complex biological matrices. Additionally, the integration of green chemistry principles, especially the use of eco-friendly solvents, is driving sustainability in analytical practices. The aim of this review is to comprehensively evaluate and synthesize recent advances in analytical methodologies for the quantification of Gentamicin across pharmaceutical formulations and biological matrices. The main objective of this study includes assessing traditional and modern analytical techniques, emphasizing methods with superior sensitivity and specificity such as HPLC-MS/MS, highlighting environmentally sustainable approaches in line with green chemistry principles.
