<p>Amber is a fossilized natural resin that has attracted growing attention due to its complex chemical structure and potential biomedical value. In this study, chemical and antibacterial fingerprinting of Baltic and Dominican amber extracts were conducted using Gas Chromatography–Mass Spectrometry (GC–MS), Fourier Transform Infrared Spectroscopy (FTIR), and standard antibacterial assays. The GC–MS analysis revealed a distinctive composition between the two types of amber. Baltic amber was found to be rich in succinic acid, valeric acid derivatives, terpenoids (such as borneol and camphor), and resin acids (isopimaric and dehydroabietic acids), whereas Dominican amber contained mainly long-chain fatty acid methyl esters dominated by methyl oleate. FTIR spectra supported these findings, confirming stronger O–H and C = O absorption bands in Baltic amber, indicative of carboxylic and hydroxyl functional groups, while Dominican amber showed pronounced aliphatic and ester peaks. The antibacterial activity, tested against Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae, demonstrated that Baltic amber extracts exhibited significantly larger inhibition zones than Dominican amber. Ethanol proved to be a more effective extraction solvent than DMSO, yielding higher antibacterial activity due to its enhanced solubility for bioactive terpenoids and resin acids. These findings establish a clear relationship between the chemical composition and antibacterial efficacy of amber, highlighting Baltic amber as a promising candidate for future biomedical and pharmaceutical applications.</p> Graphical Abstract <p></p>