METHOD FOR QUANTITATIVE DETERMINATION OF AMIODARONE IN HUMAN BLOOD PLASMA Russian patent published in 2025 - IPC G01N33/94 G01N33/49 G01N30/02 G01N30/06 

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to cardiology, and can be used for quantitative determination of amiodarone in human blood plasma by liquid chromatography. Peripheral venous blood is sampled from the patient taking amiodarone and centrifuged, and the supernatant is transferred to eppendorf to obtain blood plasma. Sample preparation is carried out using a modified method QuEChERS - Quick, Easy, Cheap, Effective, Rugged and Safe, which consists in preparation of a working calibration solution, for which 450 mcl of blood plasma is collected, then successively adding to plasma 50 mcl of amiodarone solution with concentration of 0.0125 mg/ml – average therapeutic concentration, 0.33±0.04 g of sodium chloride. Acetonitrile 500 mcl is added to the obtained solution, the mixture is centrifuged and the supernatant is collected. To prepare the analysed sample, 500 mcl of blood plasma is collected, further, 8 weights of sodium chloride are added to plasma – 0.33±0.04 g, 500 mcl of acetonitrile, centrifuged, supernatant is collected. Analysed and calibration samples are then chromatographed under gradient elution conditions using a Tsunami C18 Pharm column at a flow rate of 1.2 ml/min, an injection volume of 80 mcl, a detection wavelength of 241 nm with a total analysis time of 18 minutes. Amiodarone content in the patient's blood plasma is determined by plotting a calibration curve in the concentration range from 40% to 200% of the amiodarone concentration of 0.0125 mg/ml.

EFFECT: method enables simple, cost-effective and time-efficient determination of human plasma amiodarone concentration for a controlled approach to therapy with increased effectiveness and safety in patients with cardiac arrhythmias due to peculiarities of sample preparation based on QuEChERS method, providing elimination of significant effect of plasma proteins on chromatographic analysis, and specificity of chromatographic conditions of gradient elution, which confirms the possibility of quantitative determination of amiodarone by high-performance liquid chromatography-ultrafiltration without using a mass-spectrometric detector.

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