Development and Optimization of HPLC Conditions for Quantitative Determination of Methadone in Hospital-Prepared Syrup Formulation
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Abstract
Reliable and practical analytical methods are essential for confirming the quality and safety of locally compounded methadone syrup in clinical settings. This study aimed to develop and optimize high-performance liquid chromatography (HPLC) conditions for the quantitative determination of methadone in hospital-compounded formulations. Five distinct mobile phase compositions, varying in buffer systems and organic solvent ratios, were systematically evaluated using a reversed-phase C18 column. Chromatographic consistency was assessed based on retention time, peak area, signal reproducibility, and peak symmetry. Among the evaluated conditions, a mobile phase consisting of 20 mM ammonium formate and acetonitrile (13:87, v/v) demonstrated the most favorable chromatographic performance. The optimized condition yielded an ideal retention time of approximately 6.0 minutes, symmetrical peak profiles (As10 ≈ 1.25-1.45), and very low variability (%RSD < 3%), indicating consistent chromatographic response under the tested conditions. Additionally, the method provided a stable signal intensity with no major observable interfering peaks. This optimized framework was successfully applied to quantify methadone in hospital- prepared syrup samples. The developed methodology is streamlined, cost-effective, and highly suitable for routine implementation into hospital laboratory workflows. Overall, this study establishes a practical analytical framework for the quality control of methadone formulations in clinical environments.
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