18 / 19
allied
academies
Journal of Pharmacology and Therapeutic Research
Volume 1 Issue 1
Clinical Pharmacy 2017
Notes:
Page 58
December 07-09, 2017 | Rome, Italy
7
th
World Congress on
Clinical Pharmacy and Pharmacy Practice
Simultaneous determination of Irbesartan
and hydrochlorothiazide in human plasma
using HPLC coupled with tandem mass
spectrometry: Application to bioequivalence
studies
Lara Tutunji
University of Jordan, Jordan
A
sensitive,
specific
and
selective
liquid
chromatography/tandem mass spectrometric method
has been developed and validated for the simultaneous
determination of Irbesartan and hydrochlorothiazide in
human plasma. The chromatographic conditions were
optimized to achieve high resolution, and peak symmetry
with a short retention time for both analytes and the
internal standard. Plasma samples were prepared using
protein precipitation with acetonitrile, the two analytes
and the internal standard losartan were separated on a
reverse phase C.sub.18 column (50 mm x 4 mm, 3 [mu]
m) using water with 2.5% formic acid, methanol and
acetonitrile (40:45:15, v/v/v (%)) as a mobile phase (flow
rate of 0.70 mL/min). Irbesartan and hydrochlorothiazide
were ionized using ESI source in negative ion mode,
prior to detection by multiple reaction monitoring (MRM)
mode while monitoring at the following transitions: m/z
296→269 and m/z 296→] 205 for hydrochlorothiazide,
427→175 for Irbesartan. Linearity was demonstrated over
the concentration range 0.06-6.00 µg/mL for Irbesartan
and 1.00-112.00 ng/mL for hydrochlorothiazide. The
method demonstrated high calibration sensitivity (0.2537
and 0.0129 for Irbesartan and HCTZ, respectively). The
developed and validated method was successfully applied
to a bioequivalence study of Irbesartan (300 mg) with
hydrochlorothiazide (12.5 mg) tablet in healthy volunteers
(N=36).
tutunji@ju.edu.joJ Pharmacol Ther Res 2017