Determination of antibiotic anisomycin in tissue samples by liquid chromatography–tandem mass spectrometry

Abstract

The aim of this work was development of sensitive analytical method for determination of drug anisomycin in different tissue samples. The optimized extraction procedure included methanol as extraction solvent and Oasis HLB as extract clean up cartridge. In the method validation, ani-somycin was efficiently extracted from most of tissues, such as heart, brain, spleen, kidney and li-ver, whereas lower recoveries were obtained for fat tissue. Low limits of detection (0.6–3.8 ng g–1) and quantification (2.4–11.8 ng g–1) indicate that developed method is suitable for determination of anisomycin trace levels. The method was linear (R2 > 0.975) in the tested concentration range (50–2500 ng g–1). Significant matrix effect was eliminated using matrix-matched standards. It was finally determined that the developed method is sensitive and reliable for determination of drug anisomycin in various tissue samples using liquid chromatography–tandem mass spectrometry.

Cilj ovog rada bio je razvoj osetljive analitičke metode za određivanje leka anizomicina u različitim tkivima. U optimizovanoj proceduri ekstrakcije, metanol je odabran kao rastvarač za ekstrakciju, a Oasis HLB kao kertridž za prečišćavanje ekstrakta. Prilikom validacije razvijene metode, anizomicin je efikasno ekstrahovan iz većine tkiva, kao što su srce, mozak, slezina, bubrezi i jetra, dok su za masno tkivo dobijeni niži prinosi. Niske granice detekcije (0,6–3,8 ng g –1 ) i kvantifikacije (2,4–11,8 ng g–1 ) ukazuju da je razvijena metoda pogodna za određivanje tragova anizomicina. U ispitivanom opsegu koncentracija (50–2500 ng g–1 ) metoda je linearna (R2 > 0,975). Značajan uticaj matrice eliminisan je upotrebom standarda koji odgovaraju matrici uzorka. Na osnovu rezultata validacije utvrđeno je da je razvijena metoda osetljiva i pouzdana za određivanje leka anizomicina u različitim tkivima korišćenjem tečne hromatografije sa tandem masenom spektrometrijom.