Friday, 17 March 2017

The Rapid and Sensitive Hplc-Ms/Ms-Method of Determination of Mebeverine Metabolites in Human Plasma




The rapid and sensitive method for quantifying mebeverine metabolites in human plasma was developed using HPLC-MS/ MS. Mebeverine acid-D5 and desmethyl mebeverine acid-D5 were used as internal standards. Sample preparation was performed by protein precipitation. The chromatographic separation was achieved on Phenomenex Luna C8 Mercury (20 x 4.0 mm, 5 μm) and Phenomenex Luna 5u C8 (150 x 4.6 mm, 5 μm) under gradient conditions of mobile phase. The calibration range of mebeverine acid and desmethyl mebeverine acid was 10 – 2000 ng/ml. The method was applied to conduct a pharmacokinetic study of 200 mg prolonged-release capsules Duspatalin on 24 healthy participants.

Mebeverine is myotropic antispasmodic agent which effect is based on blockade of fast sodium channels and slow calcium channels on membrane of myocytes. It slows down depolarization of the membrane and prevents contraction of muscle fibers. The drug has high selectivity to smooth muscles of the gastrointestinal tract. Therefore it is primarily used to relieve spasms of organs of the digestive system. Mebeverine being an ester is rapidly hydrolyzed in step of first-pass elimination by esterases to 3.4-dimethoxybenzoic (veratric) acid and mebeverine alcohol. The main metabolites of the drug are mebeverine acid (MA) and desmethyl mebeverine acid.

The half-life of DMA after administration of prolonged-release capsules of mebeverine is 5-6 h, maximum concentration in blood (Cmax) after a single dose is 679 ng/ml, after repeated doses 804 ng/ml, time-to-peak concentration (Tmax) is about 2.92 hours. The area under the curve “concentrationtime” from last blood sampling procedure (AUC0-t) for DMA is 4552 ng∙h/mL, the elimination constant is 0.147. The values of pharmacokinetic parameters for MA were not published. Therefore, obtaining new data about pharmacokinetic of mebevere in the form of prolonged-release capsule is actual. Several analytical methods of quantitative determination of mebeverine metabolites in plasma using HPLC and HPLC-MS/ MS were reported. However, these methods have some disadvantages, such as necessity of complex sample preparation, using solid-phase and liquid-liquid extraction, and lack of sensitivity (high LLOQ).  

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