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Simultaneous Quantification of Pantoprazole and Levosulpiride in Spiked Human Plasma Using High Performance Liquid Chromatography Tandem Mass Spectrometry

[ Vol. 15 , Issue. 1 ]

Author(s):

Vulli Srinandan, Krishnaveni Nagappan*, Sonam Patel, Karthik Yamjala, Gowramma Byran and Basavan Babu   Pages 17 - 23 ( 7 )

Abstract:


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 mode in positive 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 0.3-2400 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.

Keywords:

Pantoprazole, levosulpiride, bioanalytical method, validation, LC-MS/MS.

Affiliation:

Department Of Pharmaceutical Analysis, JSS College of Pharmacy, [A Constituent College of Jagadguru Shree Shivarathreeshwara University, Mysuru], Udhagamandalam, Department Of Pharmaceutical Analysis, JSS College of Pharmacy, [A Constituent College of Jagadguru Shree Shivarathreeshwara University, Mysuru], Udhagamandalam, Department Of Pharmaceutical Analysis, JSS College of Pharmacy, [A Constituent College of Jagadguru Shree Shivarathreeshwara University, Mysuru], Udhagamandalam, Department Of Pharmaceutical Analysis, JSS College of Pharmacy, [A Constituent College of Jagadguru Shree Shivarathreeshwara University, Mysuru], Udhagamandalam, Department Of Pharmaceutical Chemistry, JSS College of Pharmacy, [A Constituent College of Jagadguru Shree Shivarathreeshwara University, Mysuru], Udhagamandalam, Department Of Pharmaceutical Analysis, JSS College of Pharmacy, [A Constituent College of Jagadguru Shree Shivarathreeshwara University, Mysuru], Udhagamandalam



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