TITLE:
Development of Ultra Fast Liquid Chromatography (UFLC) Method for Fluorescence Detection of Domperidone in Human Serum and Application to Pharmacokinetic Study
AUTHORS:
Chinna Reddy Palem, Satyanarayana Goda, Narendar Reddy Dudhipala, Madhusudan Rao Yamsani
KEYWORDS:
Domperidone, Ultra Fast Liquid Chromatography, Fluorescence, Human Serum, Buccal Patches, Pharmacokinetics
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.7 No.1,
January
19,
2016
ABSTRACT: A simple and sensitive
fluorescence detection of domperidone by ultra fast liquid chromatographic method
was developed and validated in human serum. For the evaluation of new drug delivery
systems, conducting of pharmacokinetic studies in human volunteers is essential
for approval to marketing after preclinical evaluation in animal models. The present
method consists of protein precipitation, extraction of analytes from human serum
into dichloromethane and separation using reversed-phase C18 column.
Propranolol hydrochloride was used as an internal standard and the eluent was monitored
by fluorescence detector at excitation 282 and emission 328 nm. The mobile phase
used was 62:38 ratio of 10 mM phosphate buffer pH adjusted to 3.1 with OPA and methanol
at a flow rate of 1 mL·min-1. The method was evaluated for assay, LLOD, LLOQ, recovery
and stability studies. The retention times for domperidone and propranolol hydrochloride
were found to be 6.36 and 7.94 minutes respectively. The intraday and inter-day
coefficient of variation and percent error values of assay method were less than
5%; mean recovery was more than 96% for each analyte and the method was found to
be precise, accurate and specific during study. The method was successfully applied
for pharmacokinetic study of immediate and controlled release bioadhesive hot melt
extruded buccal patches of domperidone after buccal administration to healthy human
volunteers. The Cmax, Tmax, and AUC0–24 of domperidone
from immediate and controlled release buccal patches were found to be 129.7 ng·mL-1, 1.5 h, 455.1 ng·h·mL-1 and 145.7 ng·mL-1, 5.25
h, 911.0 ng·h·mL-1 respectively. A simple, sensitive and reliable method for
the fluorescence determination of domperidone in human serum by UFLC method was
developed and validated.