• Nurmeilis
  • Yahdiana Harahap Faculty of Pharmacy, Universitas Indonesia, Jakarta, Indonesia
  • Fadlina Chany Saputri Universitas Indonesia
  • Abdul Munim Universitas Indonesia
  • Rianto Setiabudy Universitas Indonesia


Validated method, LC-MSMS, Drug interaction, Irbesartan, Orthosiphon stamineus


Objective: To determine and validate of irbesartan and sinensetin simultaneously by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and application of this method to study a pharmacokinetic interaction of irbesartan and ethanol extract of Orthosiphon stamineus herba in rat plasma.

Methods: The irbesartan and sinensetin were simultaneously extracted from plasma by protein precipitation with acetonitrile. Samples containing irbesartan and sinensetin were analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with C18 column Acquity® (100 mm × 2.1 mm), 1.7 μm particle size column at 40 °C. The gradient system of mobile phase composition was a mixture of acetonitrile and 0.1% formic acid (40:60 v/v), which was pumped at a flow rate of 0.3 ml/min. Mass detection was performed on Waters Xevo Triple Quadrupole equipped with an electro spray ionization (ESI) source at positive ion mode in the multiple reaction monitoring (MRM) mode. Irbesartan was detected at m/z 429.1>205.9, sinensetin was detected at m/z 373>342.9 and losartan as an internal standard was detected at m/z 423.05>404.9.

Results: The method was validated according to EMEA guidelines which showed good reproducibility and linearity of 0.99, the lower limit of quantification (LLOQ) were 25 ng/ml and 250 ng/ml for irbesartan and sinensatin, respectively. The precision (% CV) values of within-run and between-run analysis is 9.3-5.25% and 1.52–5.47% (for irbesartan), 1.52–5.09% and 2.47–9.14% (for sinensetin) whereas the accuracy (% diff) of both irbesartan and sinensetin were less than 20%. Stability studies revealed that irbesartan and sinensetin have been stable for 24 h at room temperature, 24 h in the autosampler, 3 freeze-thaw cycles, and at least 30 d at-20 oC. The validated method was applied to evaluate pharmacokinetic interactions of irbesartan and ethanol extract of Orthosiphon stamineus herba in rat plasma.

Conclusion: The developed LC/MS-MS method is valid to evaluate irbesartan and sinensetin simultaneous in vitro and showed good selectivity, linearity, accuracy, precision, matrix effect, and stability. The method was successfully applied to study the pharmacokinetics interaction of irbesartan and Orthosiphon stamineus herba in rat plasma.



Download data is not yet available.


Charles FL, Lora LA, Morton PG, Leonard LL. Drug Information Handbook, A Comprehensive Resource for all Clinicians and Healthcare Professionals, 19th edition, New York: Lexi-Comp Inc; 2011. p. 832-4.

Siddiqui MJ, Ismail Z. Simultaneous analysis of bioactive markers from Orthosiphon stamineus Benth leaves extracts by reverse phase high-performance liquid chromatography. Trop J Pharm Res 2011;10:97-103.

AdnyanaI K, Setiawan F, Insanu M. From ethnopharmacology to clinical study of Orthosiphon stamineus Benth. Int J Pharm Sci 2013;5:66-73.

Patel B, Jivani NP, Khodakiya A. Drug interaction: can we make them advantageous for a human being? Int J Pharm Res Dev 2012;4:8-15.

Ganesan M, Nanjundan S, Gomathi M, Muralidharan S. Method development and validation of irbesartan using LC-MS/MS: application to pharmacokinetic studies. J Chem Pharm Res 2010;2:740-6.

Harmita, Harahap Y, Arya IK. Validation of analitycal method of irbesartan plasma in vitro by high-performance liquid chromatography-fluorescence. J Life Sci 2012;6:726-31.

European Medicines Agency (EMEA), Committee for Medicinal Products for Human Use (CHMP). Guideline on bioanalytical method validation. London; 2011.

Evans G. A handbook of bioanalysis and drug metabolism. Boca Rato, Florida: CRC Press; 2004.

Hanapi NA, Azizi J, Ismail S, d and Mansor SM. Evaluation of selected Malaysian medicinal plants on phase I drug metabolizing enzymes CYP2C9, CYP2D6, and CYP3A4 activities in vitro. Int J Pharm 2010;6:1-6.

Agilent Technologies. Basics of LS-MS, U. S. A 5988-2045EN; 2011. Available from: http// wipf/ Agilent % 20LC-MS%20Primer.pdf. [Last accessed on 10 Jul 2015].

Husain A, Bhasin PS, Nagar H. A review of the pharmacological and pharmaceutical profile of irbesartan. Pharmacophore 2011;2:276–86.



How to Cite

Nurmeilis, Y. Harahap, F. C. Saputri, A. Munim, and R. Setiabudy. “DETERMINATION OF IRBESARTAN AND SINENSETIN SIMULTANEOUSLY BY LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY AND THEIR APPLICATION TO DRUG INTERACTION STUDY IN RAT PLASMA”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 8, no. 1, Jan. 2016, pp. 96-100,



Original Article(s)