T. Sudha, Kalan Kumar Reddy, P. V. Hemalatha, V. R. Ravikumar


A sensitive and selective liquid chromatographic method coupled with tandem mass spectroscopy (LC-MS/MS) was developed for the quantification of omeprazole in human plasma. Lansoprazole was used as internal standard with plasma samples, extracted using 10mM ammonium acetate. A centrifuged upper layer was then evaporated and reconstituted with Acetonitrile: mobile phase buffer 70:30%v/v. The reconstructed samples were injected into a C18 column purospher star 5µ. The mobile phase was composed of ACN: mobile phase buffer (5mm ammonium bicarbonate buffer) in the ratio of 70:30%v/v with flow rate 1.0mL/min. The mass spectrometer was operated using positive ion mode and turbo electro spray ionisation. Nitrogen was used as the nebulizer, curtain, collision and auxiliary gases. Using MS/MS with multiple reactions monitoring (MRM) mode, omeprazole was detected without severe interferences from plasma matrix. Detection of omeprazole in human plasma was accurate and precision. This method has been successfully applied to the study of omeprazole in human specimens

Keywords: Proton pump inhibitor, omeprazole, lansoprazole, LC-MS/MS, liquid liquid  extraction

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Antonio Checa, Ramon Oliver, Santiago Hernandez Cassou, Javier Saurina, 2009. Determination of HIV drugs in biological matrices: A review. Anal. Chimica Acta.647:1-13.

Bharathi DV., Hotha KK., Jagadeesh B., Chatki PK., Thriveni K., Mullangi R, Naidu A., 2009. Simultaneous estimation of four proton pump inhibitors--lansoprazole, omeprazole, Pantoprazole and rabeprazole development of a novel generic HPLC-UV method and its application to clinical pharmacokinetic study. J. Bio. Med. chromatogr. 23(7): 732 - 739.

Dams R., Huestis MA., Lambert WE., Murphy CM., 2003. Matrix effect in Bio-analysis of liquid drugs with LC-MS/MS: influence of ionization type, sample preparation and bio fluid. J. Am. Soc. Mass. 14(11):1290-94.

De Smet J., Boussery K., De Cock P., De Paepe P., Remon JP., Van Winckel M., Van Boussery J., 2010. A Bio-analytical hydrophilic interaction LC- MS/MS method for the simultaneous quantification of omeprazole and lansoprazole in human plasma in support of a pharmacokinetic omeprazole study in children. J. Sep. Sci. 33(6-7): 939-947.

Furuta T., Takashima M., Shirai N., Xiao F., Hanai H., Ohashi K., Ishizaki T., 2000. Cure of refractory duodenal ulcer and infection caused by Helicobacter pylori by high doses of omeprazole and amoxicillin in a homozygous CYP2C19 extensive metabolizer patient. Clin. Pharmacol. Ther. 67: 684 – 689.

Guidance for Industry: ICH E6 Good Clinical Practice, US Department of Health and Human services, Food and Drug Administration Centre for Drug Evaluation and Research (CDER), Centre for Biologics Evaluation and Research (CBER), April 1996.

Guidance for Industry, Bio analytical method validation, US Department of Health and Human services, Food and Drug Administration Centre for Drug Evaluation and Research (CDER), Centre for Veterinary Medicine(CBER), May 2001.

Jian Liu, Jian-zhong Shentu, Li-hua Wu, Jing Dou, Qi-yang Xu, Hui-li Zhou, Guo-lan Wu, Ming-zhu Huang, Xing-jiang Hu, Jun-chun Chen.2012. Relative bioavailability and pharmacokinetic comparison of two different enteric formulations of omeprazole. J. Zhejiang. Univ. Sci B. 13(5): 348–355.

Kamrun Nahar, Jafreen Jamal Joti, Md Ashik Ullah, Ahasanul Hasan, Mohammad Abul Kalam Azad, Abul Hasnat, 2009.A Simple RP-HPLC Method for the Determination of Omeprazole in Human Serum and Urine: Validation and Application in Pharmacokinetic Study. Dhaka University J. Pharma. Sci. 8(2):

Kanazawa H., Okada A., Matsushima Y., Yokota H., Okubo S., Mashige F., Nakahara K., 2004. Determination of omeprazole and its metabolites in human plasma by liquid Chromatography-mass spectrometry. J. Chromatography. 949(1-2): 1-9.

Kang WK., Kim DS., Kwon KI., 1999. Advanced method for determination of omeprazole in plasma by HPLC. Arch. Pharma. research. 22(1); 86-88.

Matuszewski BK., Constanzer ML., Chavez-Eng CM., 2003. Strategies for the Assessment of Matrix effect in Quantitative Bio analytical methods based on HPLC- MS/MS. J Anal. Chem. 75(13):3019-30.

Rambla-Alegre M., Esteve-Romero J., Carda-Broch S.. 2009. Analysis of omeprazole and its main metabolites by liquid chromatography using hybrid micellar mobile phases. Anal. Chim. Acta. 633(2):250-6

Rang HP., Dale MM., Ritter JM., Flower RJ. 2007. Pharmacology. 6thed. Elsevier, Philadelphia, Churchill Livingstone, pp. 295-96.

Stenhoff H., Blomqvist A., Lagerström PO.,1999. Determination of the enantiomers of omeprazole in blood plasma by normal-phase liquid chromatography and detection by atmospheric pressure ionization tandem mass spectrometry. J. Chromatogr. B. 734: 191.

Shiohira H., Yasui-Furukori N., Tateishi T., Uno T..2011. Chiral assay of omeprazole and metabolites and its application to a pharmacokinetics related to CYP2C19 genotypes. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 879(24):2465-70

Taylor PJ., 2005. Matrix effects: the Achilles heel of quantitative high-performance liquid chromatography-electrospray-tandem mass spectrometry. Clin Biochem.38(4):328-34

TsingHua. 2004. Determination of relative bioavailability of omeprazole enteric-coated capsules in healthy volunteer. Chinese.J. hospital pharmacy. 2002 - 08.

Vittal S., Ganneboina R., Layek B., Rived RK., Hotha KK., Bharathi DV., Mullangi R., 2009. Highly sensitive method for the determination of omeprazole in human plasma by liquid Chromatography-electrospray ionization tandem mass spectrometry: application to a Clinical pharmacokinetic study. Biomed. chromatogram. 23(4): 390-6.

Wang J., Wang Y., Fawcett JP., Wang Y., Gu J. 2004. Determination of omeprazole in human plasma by liquid chromatography – electrospray quadrupole linear ion trap mass spectrometry. J. Bio med anal. 39 (3-4): 631-635.

Woolf EJ., Matuszewski BK., 1998. Simultaneous determination of omeprazole and 5'-s hydroxyomeprazole in human plasma by liquid chromatography-tandem mass spectrometry. J. Chromatogr A 828(1-2): 22.

Yim DS., Jeong JE., Park JY., 2001. Assay of omeprazole and omeprazole sulfone by semi-microcolumn liquid chromato-graphy with mixed-function precolumn. J Chromatogr B Biomed Sci Appl.754(2):487-93

Zarghi A., Foroutan SM., Shafaati A., Khoddam A., 2006. Determination of omeprazole in blood plasma by using HPLC. Arzneimittelfor schung 2006;56(6) 382-6.



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