Validated method for determination of bromopride in human plasma by liquid chromatography--electrospray tandem mass spectrometry: application to the bioequivalence study.
A simple, sensitive and specific liquid chromatography-tandem mass spectrometry method for the quantification of bromopride I in human plasma is presented. Sample preparation consisted of the addition of procainamide II as the internal standard, liquid-liquid extraction in alkaline conditions using hexane-ethyl acetate (1 : 1, v/v) as the extracting solvent, followed by centrifugation, evaporation of the solvent and sample reconstitution in acetonitrile. Both I and II (internal standard, IS) were analyzed using a C18 column and the mobile-phase acetonitrile-water (formic acid 0.1%). The eluted compounds were monitored using electrospray tandem mass spectrometry. The analyses were carried out by multiple reaction monitoring (MRM) using the parent-to-daughter combinations of m/z 344.20 > 271.00 and m/z 236.30 > 163.10. The areas of peaks from analyte and IS were used for quantification of I. The achieved limit of quantification was 1.0 ng/ml and the assay exhibited a linear dynamic range of 1-100.0 ng/ml and gave a correlation coefficient (r) of 0.995 or better. Validation results on linearity, specificity, accuracy, precision and stability, as well as application to the analysis of samples taken up to 24 h after oral administration of 10 mg of I in healthy volunteers demonstrated the applicability to bioequivalence studies.
Nazare
P
P
Clinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School, Av. Sao Francisco de Assis 218, 12916-900 Bragança Paulista, SP, Brazil.
Massaroti
P
P
Duarte
L F
LF
Campos
D R
DR
Marchioretto
M A M
MA
Bernasconi
G
G
Calafatti
S
S
Barros
F A P
FA
Meurer
E C
EC
Pedrazzoli
J
J
Moraes
L A B
LA
D002851:Chromatography, High Pressure Liquid; D018492:Dopamine Antagonists / Q000097:blood* / Q000493:pharmacokinetics*; D006801:Humans; D008787:Metoclopramide / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013810:Therapeutic Equivalency
D018492:Dopamine Antagonists; C013855:bromopride; D008787:Metoclopramide