High-performance liquid chromatographic method for the determination of moclobemide and its two major metabolites in human plasma

A validated high‐performance liquid chromatographic method for the determination of moclobemide and its two metabolites in human plasma and application to pharmacokinetic studies

“…Geschke et al (1987) extracted moclobemide and its metabolites on the Extrelut 1 glass columns packed with keiselguhr (natural diatomaceous earth); however, the cost of these columns is high. More economical columns were employed for sample preparation, such as Bakerbond CN (Misztal et al, 2002), Speedisk ® H 2 0-Philic DVB (Rakic et al, 2006) and Bond Elut C 18 cartridges. These procedures are, however, time-consuming (Hoskins et al, 2001a).…”

“…These procedures are, however, time-consuming (Hoskins et al, 2001a). For the quantitative measurement of the analytes Spherisorb C 6 (Geschke et al, 1987), Nova-Pak C 8 (Misztal et al, 2002), Waters Xterra™RP18 (Rakic et al, 2006) and Nova-pak phenyl columns (Hoskins et al, 2001a) with the mobile phase containing acetonitrilie and 0.067 M potassium phosphate buffer adjusted to pH 3.9 with hydrochloric acid (Geschke et al, 1987) or acetonitrile and 0.2 M ammonium formate buffer pH 3.57 (Hoskins et al, 2001a) have been employed. The mobile phase used by Misztal et al (2002) was more complicated.…”

“…It composed with acetonitrile, methanol, 0.067 M phosphate buffer pH 2.65 and 0.4% triethylamine used for the improvement of peak shape of analyzed substances. The same mobile phase additive, triethylamine, at a concentration of 1% (pH 3.9) was used in the mobile phase of acetonitrile and 10 mM KH 2 PO 4 (83:17, v/v; Rakic et al, 2006). The declared limits of quantification (LOQ) for moclobemide Ro 12-8095 and Ro 12-5637 were in range 10-30 ng/mL for the methods with UV detection (Geschke et al, 1987;Misztal et al, 2002;Rakic et al, 2006), and 10 ng/mL in HPLC-ESI-MS method (Hoskins et al, 2001a).…”

“…This metabolite is formed by N-oxidation of the morpholine moiety, but the reaction is mediated rather by FMO than the cytochrome P450 (Hoskins et al, 2001b). Ro 12-5637 is a weak inhibitor of MAO-A. To the best of our knowledge, only few analytical techniques have been developed for the determination of moclobemide and its metabolites in biological fluids (Geschke et al, 1987;Misztal et al, 2002;Hoskins et al, 2001a,c;Ponds et al, 1990;Rakic et al, 2006). Highperformance liquid chromatography (HPLC) with ultraviolet detection (Geschke et al, 1987;Misztal et al, 2002;Rakic et al, 2006) and mass spectrometry (Hoskins et al, 2001a) have been reported for monitoring plasma concentrations of moclobemide and its metabolites.…”

“…The overall chromatographic run time was 10 min. The short time allowed for more rapid sample throughput than the published liquid chromatography techniques with UV detection (Geschke et al, 1987;Misztal et al, 2002;Rakic et al, 2006).…”