LC–MS/MS assay
method development and validation for the simultaneous quantification of Solifenacin
and Mirabegron in human plasma
Reddy Penchala Vaka and Battula Sreenivasa Rao
Res. J. Chem. Environ; Vol. 26(6); 115-122;
doi: https://doi.org/10.25303/2606rjce115122; (2022)
Abstract
The present study intended to develop a simple and novel Liquid Chromatography–Mass
Spectrometry/ Mass spectrometry (LC–MS/MS) method for the simultaneous quantification
of solifenacin and mirabegron, a combined medication used for the treatment of overactive
bladder symptoms in human plasma. Tolterodine drug is used as an internal standard
in the study. Both the analytes and internal standard were isolated from 100 μL
plasma samples by liquid-liquid extraction and then chromatographed on Zorbax C18
(4.6 mm×50 mm, 5 μm) column with a mobile phase consisting of methanol and 5mM ammonium
formate in the ratio of 25:75 (v/v) pumped at 0.3 mL/min. The method had a chromatographic
total run time of 5 min. The developed method gave symmetric peak at a retention
time of 2.50 min, 2.99 min and 1.18 min respectively for solifenacin, mirabegron
and nifedipine satisfying all the peak properties as per USP guidelines. The mass
spectral characterization of separated analytes in the LC method was performed using
mass detector operated at Multiple Reaction Monitoring mode with precursor-toproduct
ion transitions at m/z of 363 to m/z of 110 as MH+ ion for solifenacin, m/z of 397
to m/z of 239 as MH+ ion for mirabegron. A very sensitive limit of detection of
7.5 ng/mL was observed and showed calibration curve linear over the concentration
range of LLOQ to 1000 ng/mL.
The other validation parameters were found to have acceptable accuracy, precision,
linearity and selectivity. The mean extraction concentration was acceptable and
very high for both the analytes in HQC, MQC and LOQ levels. Various stability studies
of solifenacin and mirabegron such as freeze-thaw, short term, long term, auto-sampler
and dry extract stability proved that the method was stable. Based on the results,
it can be proved that the method was accurate, precise and specific for the simultaneous
analysis of solifenacin and mirabegron in human plasma.
