Development and validation of LC-MS/MS method for the determination of amikacin in human plasma and its application in adult hospitalized patients in Yogyakarta Indonesia

Abstract

Therapeutic drug monitoring (TDM) is critical to ensure the safe and effective administration of amikacin (AMK), which has a narrow therapeutic index. This study aimed to develop and validate an efficient and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determining AMK in human plasma and applying it to hospitalized patients. Before LC-MS/MS measurements, plasma samples were treated with an optimized protein precipitation method using methanol (MeOH). Optimum chromatographic conditions for determining AMK in human plasma include using a C8 column (5 μm, 100 × 4.6 mm) and gradient elution using ammonium formate-formic acid-H2O and formic acid-MeOH mixture as the mobile phase at a 0.5 ml/minute flow rate. The method has been validated following the European Medicines Agency guidelines and has met all validation requirements with excellent selectivity, an lowest limit of quantification value of 0.2 μg/ml, good accuracy and precision in the linear range of 0.2–25 μg/ml, good dilution integrity up to 20 dilution times, and stable under various conditions. The method has also successfully been applied for measuring AMK peak levels (Cmax) and trough levels (Cmin) in the plasma of adult inpatients at Dr. Sardjito Hospital Yogyakarta, Indonesia. The LC-MS/MS measurement can be completed in 7.5 minutes, and the method can be applied in the TDM of AMK.