Open AccessHPLC-PDA METHOD FOR THE QUANTIFICATION OF PARACETAMOL IN PLASMA: APPLICATION TO PK/PD STUDIES WITH ARTHRITIC RATS
Author(s) -
Aldo Bazán Ramírez,
Francisco López-Múñoz,
J. Medina,
Marcela Hurtado,
Georgina Alarcón Ángeles,
Adriana Pineda,
Luis Alfonso Moreno-Rocha
Publication year - 2017
Publication title -
international journal of pharmacy and pharmaceutical sciences/international journal of pharmacy and pharmaceutical sciences
Language(s) - English
Resource type - Journals
eISSN - 2656-0097
pISSN - 0975-1491
DOI - 10.22159/ijpps.2017v9i5.17746
Subject(s) - chromatography , high performance liquid chromatography , chemistry , pharmacokinetics , protein precipitation , detection limit , pharmacodynamics , mass spectrometry , pharmacology , medicine
Objective: To develop and validate an easy, rapid, sensitive and selective high-performance liquid chromatography with photodiode diode-array (HPLC-PDA) detection method for quantification of paracetamol and to demonstrate its application in a pharmacokinetic–pharmacodynamic study with arthritic rats.Methods: Paracetamol was separated from plasma samples (50-100 µl) by a single protein precipitation step, prior to HPLC-PDA detection. The separation was performed on a Knauer Eurospher II, C18 column 5 µm, 150 × 4.6 mm. The mobile phase comprised a mixture of water: methanol (75:25) and the flow rate was 1.1 ml/min. The detection wavelength was set at 245 nm. All analyses were carried out at room temperature (25 °C). Pharmacodynamics data were obtained with a gout-type pain model in rats.Results: The method was linear within a range of 0.2-200 µg/ml (R2≥0.99). The intra-day and inter-day precision and accuracy expressed as coefficient of variation and relative error, respectively were below 10%. The lower limit of quantification was 0.2 µg/ml. Plasma samples were stable at least for 5 w at ‒20° C.Conclusion: The validated method is sensitive, precise, accurate and specific as other more complex high-performance liquid chromatographic methods coupled to mass spectrometry (HPLC-MS), using small plasma samples (50-100 µl) and with a short time analysis (<5 min). The method was successfully applied to a pharmacokinetic-pharmacodynamic study of paracetamol in arthritic rats.