Liliya Logoyda, Dmytro Korobko, Oleksandra Oleshchuk, Taras Proniv, Mariya Dmutriv


Objective: A highly specific, sensitive and rapid HPLC-MS/MS method has been developed and validated for the simultaneous quantification of bisoprolol and enalapril in the present of enalaprilat in human plasma.

Methods: Analytes were extracted from plasma using a protein precipitation extraction method. Chromatography was achieved on Discovery C18, 50 × 2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A  (acetonitrile – water – formic acid, 5 : 95 : 0.1 v/v), eluent B (acetonitrile –  formic acid, 100 : 0.1 v/v)). The initial content of the eluent B is 0%, which increases linearly by 1.0 min to 100% and to 1.01 min returns to the initial 0%. The mobile phase was delivered at a flow rate of 0.400 ml/min into the mass spectrometer ESI chamber. The sample volume was 5 μl.

Results: The total chromatographic run time was 2.0 minutes and the elution of bisoprolol, enalapril, enalaprilat and IS (verapamil) occurred at ~1.01, 1.03, 0.96 and 1.09 minutes, respectively. A linear response function was established at 0.5 -50 ng/ml for bisoprolol fumarate, 2 -200 ng/ml for enalapril maleate, 1 -100 ng/ml for enalaprilat dehydrate  in human plasma. The intraday and interday accuracy and precisions were in the range of 0.311 % -0.647 % and 0.364 % - 0.572 % for bisoprolol, 0.321 % - 0.747 % and 0.390 % - 0.673 % for enalapril, 0.221 % - 0.547 % and 0.264 % - 0.773 % for enalaprilat, respectively.

Conclusion: A new rapid method was developed for simultaneous determination of bisoprolol and enalapril in the present of enalaprilat in human plasma. The method was strictly validated according to the ICH guidelines. The information thus obtained from the study can be used for the full pharmacokinetic profiling in individuals.


Yuliya Kondratova, Liliya Logoyda, Yuliia Voloshko, Ahmed Abdel-Megied, Dmytro Korobko, Yuriy Soroka. Development and validation of HPLC-DAD method for the determination of bisoprolol in tablet dosage forms. Int J Appl Pharm 2017; 9: 54-9.

Liliya Logoyda, Dmytro Korobko, Iryna Ivanusa, Kovalenko Serhii. Development of the methodology of the chromatographic determination of nifedipine in medicines. Asian J Pharm Clin Res 2017; 10: 149-152.

Dinga L, Zhoua X, Guoa X, Songa Q, He J, Xub G. LC-ESI-MS method for the determination of bisoprolol in human plasma. J Pharm Biomed Anal 2007;44:520-5.

Bhatt J, Subbaiah G, Kambli S, Shah B, Patel M, Saxena A, et al. A high throughput and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the estimation of bisoprolol in human plasma using multiplexing technique. J Chromatogr B 2007;852:374-81.

Liu GY, Wang W, Jia JY, Lu C, Liu YM, Zhang MQ, et al. Liquid chromatography tandem mass spectrometry method for determination of bisoprolol in human plasma using d5-bisoprolol as the internal standard. Biomed Chromatogr 2010;24:574-80.

Hemavathi G, Hipparagi SM. Sensitive liquid chromatography tandem mass spectrometric method for the simultaneous determination of bisoprolol and triamterene in human plasma. Asian J Pharm Clin Res 2017;10:341-8.

Yang XM, Wang CB. Determination of bisoprolol in urine by fluorometry. Fenxi Shiyanshi Chin 2001;20:54-5.

Braza AJ, Modamio P, Lastra CF, Mariño EL. Development, validation and analytical error function of two chromatographic methods with fluorimetric detection for the determination of bisoprolol and metoprolol in human plasma. Biomed Chromatogr 2002;16:517-22.

Agapova NN, Vasileva E. HPLC method for determination of bisoprolol and potential impurities. J Chromatogr A 1993;654:299-302.

Kintz P, Lohner S, Tracqui A, Mangin P, Lugnier A, Chaumont AJ. Rapid HPLC determination of bisoprolol in human plasma. J Anal Chem 1990;336:517-9.

Buehring KU, Garbe A. Determination of the new beta-blocker bisoprolol and of metoprolol, atenolol and propranolol in plasma and urine by high-performance liquid chromatography. J Chromatogr Biomed Appl 1986;55:215-24.

Harmonised Tripartite Guideline: Validation of Analytical Procedures: Methodology (Q2B) 2005. International Conference on Harmonisation (ICH); 2005.

Hernando MD, Petrovic M, Fernandez-Alba AR, Barcelo D. Analysis by liquid chromatography-electrospray ionization tandem mass spectrometry and acute toxicity evaluation for beta-blockers and lipid-regulating agents in wastewater samples. J Chromatogr A 2004;1046:133-40.

Li S, Liu G, Jia J, Liu Y, Pan C, Yu C, et al. Simultaneous determination of ten antiarrhythmics drugs and a metabolite in human plasma by liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2007;847:174-81.

Darshan Bhatt, B Rajkamal. A UPLC-MS/MS method development and validation for the estimation of sofosbuvir from human plasma. Int J Appl Pharm 2017;9:30-6.

Delma D’cruz, Anu Babu, Eena Joshy, Aneesh TP. Bioanalytical method development and validation of ticagrelor by RP-HPLC. Int J Appl Pharm 2017;9:51-4.

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