Background:
Pantoprazole (PTZ) and Levosulpiride (LS) were proven as effective agents for
the treatment of Gastro-Esophageal Reflux Disease (GERD). It is a complex motor disorder that results
in regurgitation of the gastric contents into the lower esophagus with consequent symptoms such as
heart burn, retrosternal pain, dysphagia and belching.
Methods:
A rapid, sensitive, selective and specific liquid chromatography- electro spray ionization
tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous quantification of
Pantoprazole (PTZ) and Levosulpiride (LS) in spiked Human Plasma. The method utilized SPE as sample
preparation technique and the analysis was carried out on a HPLC system utilizing electro spray
ionization as interface and triple quadrupole mass analyzer for quantification in MRM possitive mode.
Iloperidone was used as internal standard (IS). Chromatographic separation was performed on a Phenomenex
C-18 Column (4.6 mm x 50 mm, 5µ) with an isocratic elution mode utilizing a mobile phase
composition of Solution containing a mixture of 70 volumes of acetonitrile: 30 volumes of methanol
and 10mM ammonium formate (pH 4.0) at the ratio of 80:20 % v/v. The flow rate was maintained at
0.3 mL/min.
Results:
PTZ, LS and IS were detected and quantified with proton adducts at m/z 383.37→200.00, m/z
341.42→112.15 and 426.48→261.00 respectively. The linearity and range was established by fortifying
blank plasma samples in the concentration range of 3.5-2000 ng/mL for PTZ and 3.0-2400 ng/mL for
LS. The correlation coefficient (r2) was found to be ≥ 0.993 for PTZ and (r2) ≥ 0.990 for LS. The lower
limit of quantification for PTZ was 3.5 ng/mL and LS was 3.0 ng/mL. The intra and inter day precision
and accuracy for PTZ and LS were within the limits fulfilling the international acceptance criteria. PTZ
and LS were found to be stable throughout three freeze-thaw cycles, bench top and short term stability
studies.
Conclusion:
The proposed validated LC-MS/MS method offers a sensitive quantification of PTZ and
LS in spiked human plasma and can be utilized for the quantification of PTZ and LS in real-time
samples.
A new stability-indicating RP-HPLC method for the determination of dicyclomine hydrochloride and dimethicone combination in tablet dosage forms