BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS DETERMINATION OF VERAPAMIL AND ENALAPRIL IN THE PRESENT OF ENALAPRILAT BY HPLC-MS/MS


Liliya Logoyda

Abstract


Objective: The main purpose of this study was to develop a simple, precise, rapid and accurate method for the simultaneous quantification of verapamil 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 verapamil, enalapril, enalaprilat and IS (bisoprolol) occurred at ~1.09, 1.03, 0.96 and 1.01 minutes, respectively. A linear response function was established at 1 - 100 ng/ml for verapamil hydrochloride, 2 -200 ng/ml for enalapril maleate, 1 -100 ng/ml for enalaprilat dehydrate  in human plasma. The % accuracy of LLOQ samples prepared with the different biological matrix lots were found 99.2 % for enalapril, 103.4 % for enalaprilat, 90.6 % for verapamil, which were found within the range of 80.00-120.00% for the seven different plasma lots. % CV for LLOQ samples was observed as 4.9%, 5.9 %, 9.0 % respectively, which are within 20.00% of the acceptance criteria. The % mean recovery for enalapril in LQC, MQC and HQC was 103.6 %, 100.1 % and 102.4 %,  for enalaprilat in LQC, MQC and HQC was 103.2 %, 98.0 % and 101.1 %, for verapamil in LQC, MQC and HQC was 97.9 %, 96.1 % and 98.6 % respectively.

Conclusion: A new rapid method was developed for simultaneous determination of verapamil and enalapril in the present of enalaprilat in human plasma. The method was strictly validated according to the ICH guidelines. Acquired results demonstrate that proposed strategy can be effortlessly and advantageously applied for routine examination of verapamil and enalapril in the present of enalaprilat in human plasma.


References


https://pubchem.ncbi.nlm.nih.gov

https://dailymed.nlm.nih.gov

Liliya Logoyda, Dmytro Korobko, Iryna Ivanusa, Kovalenko Serhii. Development of the methodology of the chromatographic determination of nifedipine in medicines. Asian Journal of Pharmaceutical and Clinical Research. 2017; 10: 149-152.

Bhardwaj, S.P., Singh, S.; Study of forced degradation behavior of enalapril maleate by LC and LC–MS and development of a validated stability- indicating assay method Journal of Pharmaceutical and Biomedical Analysis 2008; 46: 113–120.

Ghosh, C., Jain, I., Shinde, C.P., Chakraborty, B.S. Rapid and sensitive liquid chromatography/tandem mass spectrometry method for simultaneous determination of enalapril and its major metabolite enalaprilat, in human plasma: application to a bioequivalence study; Drug Testing and Analysis 2012; 4: 94–103.

Danafar, H., Hamidi, M. Liquid chromatography–tandem mass spectrometry (LC-MS) method for the assignment of enalapril and enalaprilat in human plasma; Pharmaceutical and Biomedical Research 2015; 1: 47–58.

Guideline on Bioanalytical method validation: European Medicines Agencies; 2011.

Gu Q, Chen X, Zhong D, Wang Y. Simultaneous determination of enalapril and enalaprilat in human plasma by liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2004; 813(1-2):337-42.

Sonawane LV, Poul BN, Usnale SV. Bioanalytical method validation and its pharmaceutical application a review. Pharm Anal Acta 2014;5:1-7.

Pushpa Latha E, Sailaja B. Bioanalytical method development and validation by HPLC: a review. J Appl Pharm 2014;1:1-9.

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.

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.

Liliya Logoyda, Yuliya Kondratova, Dmytro Korobko, Yuriy Soroka. Development of UHPLC Method for the Determination of Captopril in Pharmaceutical Dosage Forms. Asian Journal of Pharmaceutical and Clinical Research. 2017; 10: 308-310.

Liliya, L., Dmytro, K., Olena, S., Ihor, B., Tamara, K. Development of methodology for identification of captopril in medicines. Asian Journal of Pharmaceuticals. 2016; 10: 168-171.

Olgya Polyauk, Liliya Logoyda. The investigation of conditions of API from group of calcium channel blockers extraction by organic solvents by using high-performance liquid chromatography as method assay. Asian Journal of Pharmaceutical and Clinical Research. 2017; 10: 354-6.

Asha B. Thomas, Ashish Naphade, Snehal S. Karanjkhele. Determination of alprazolam and fluoxetine HCl from spiked rat plasma using HPTCL with UV deterction. Int J Pharm Pharm Sci 2016;8:147-151.




Fatal error: Call to a member function getGalleyLabel() on null in /home/innowar1/public_html/journals/cache/t_compile/%%38^38D^38D7420B%%article.tpl.php on line 189