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Dose Determination of Activated Charcoal in Management of Amitriptyline-Induced Poisoning by Reversed-Phase High-Performance Liquid Chromatography
Abstract
Purpose: To assess the doses of activated charcoal currently used in the management of acute amitriptyline-induced drug poisoning and explore the possibility of using lower doses.
Methods: Albino male Wistar rats, weighing 200 ± 20 g, were used for the study. The animals were divided into four groups of eight animals each. The concentration of amitriptyline in rat plasma was measured by high performance liquid chromatography (HPLC) for dose determination of activated charcoal. Chromatograms were established with acetonitrile: 70 mM KH2PO4 buffer (60: 40, v/v) solvent system on an Xterna® ms C18 SUM column (5 μm, 3.9 × 150 mm) and pH was adjusted to 4.5 with ortho-phosphoric acid. Mobile phase flow rate was 1 ml/min and ultraviolet (UV) detection was at 293 nm. Validation of the method was performed to determine its selectivity, linearity, precision, as well as limits of detection (LOD) and of quantification (LOQ).
Results: Standard curves were linear, r2 = 0.996, for amitriptyline over the concentration range 10 - 60 ng/ml. Recovery (98.3 to 100.85 %) was in the selected concentration range of 10 - 60 ng/ml. The LOD and LOQ of the method for amitriptyline were 0.109 and 0.332 μg/ml, respectively. The validated method was successfully applied to measure plasma concentrations of amitriptyline and to measure the doses of activated charcoal currently used in the management of acute amitriptyline drug poisoning.
Conclusion: The proposed RP-HPLC method enables determination of amitriptyline with good separation and resolution of the chromatographic peaks. Validation revealed that the method is sensitive, accurate and selective. Using half of the standard dose of the activated charcoal gave a comparable effect to the standard dose in reducing drug concentration in the blood. While, using quarter of the standard dose of activated charcoal does not have a cleared effect.
Keywords: Amitriptyline, Activated charcoal, Drug poisoning, Reversed-phase high-performance liquid chromatography
