DEVELOPMENT AND VALIDATION OF LC-MS/MS METHOD FOR ESTIMATION OF UROCARB IN HUMAN PLASMA

IRYNA DRAPAK; BORYS ZIMENKOVSKY; LINA PEREKHODA; SERGIY KOVALENKO; LILIYA LOGOYDA

doi:10.22159/ijap.2019v11i5.33873

Authors

IRYNA DRAPAK General, Bioinorganic, Physical and Colloidal Chemistry Department, Danylo Halytsky Lviv National Medical University, Lviv City, Ukraine
BORYS ZIMENKOVSKY Pharmaceutical, Organic and Bioorganic Chemistry Department, Danylo Halytsky Lviv National Medical University, Lviv City, Ukraine
LINA PEREKHODA Medicinal Chemistry Department, National University of Pharmacy, Kharkiv City, Ukraine
SERGIY KOVALENKO Organic and Bioorganic Chemistry Department, Zaporizhzhya State Medical University, Zaporizhzhya City, Ukraine
LILIYA LOGOYDA Pharmaceutical Chemistry Department, Pharmaceutical Faculty, I. Horbachevsky Ternopil State Medical University, Ternopil City, Ukraine

DOI:

https://doi.org/10.22159/ijap.2019v11i5.33873

Keywords:

LC-MSMS, Urocarb, Human plasma, Validation, Pharmacokinetic studies

Abstract

Objective: The present study was aimed to develop a rapid, specific and sensitive method based on LC-MS/MS method was developed for the determination of urocarb using etomidate as an internal standard.

Methods: 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 of 8%, which increases linearly to 1.0 min to 100%, is maintained up to 1.5 min and returned to the original 8% to 1.51 min. The mobile phase was delivered at a flow rate of 0.400 ml/min into the mass spectrometer ESI chamber. The sample volume was 4 μl.

Results: The total chromatographic run time was 2.0 min and the elution of urocarb and IS (etomidate) occurred at ~1.53 and 1.67 min, respectively. A linear response function was established at 1-100 ng/ml for urocarb and etomidate in human plasma. The % mean recovery for urocarb in LQC, MQC and HQC was 104.1 %, 100.0 % and 97.4 %. The lowest concentration with the RSD<20% was taken as LLOQ and was found to be 1.03 ng/ml for urocarb. The within-run coefficients of variation ranged between 0.271 % and 0.478 % for urocarb. The within-run percentages of nominal concentrations ranged between 99.12 % and 100.21 % for urocarb. The between-run coefficients of variation ranged between 0.388 % and 0.601 % for urocarb. The between-run percentages of nominal concentrations ranged between 98.78 % and 101.11 % for urocarb.

Conclusion: A highly sensitive, specific, reproducible, rapid and high-throughput LC-MS/MS assay was developed and validated to quantify urocarb in human plasma as per the regulatory guidelines. Due to the sensitivity of the developed method, it can be applied to the monitoring of plasma levels in the analysis of drug in preclinical and clinical pharmacokinetic studies. All the parameters and results were found within the acceptance limit as given in the validation protocol.

Downloads

Download data is not yet available.

References

https://www.waters.com/webassets/cms/library/docs/720002710en.pdf. [Last accessed on 10 Mar 2019]

Claudiu T Supuran, Andrea Scozzafava, Angela Casini. Carbonic anhydrase inhibitors. Med Res Rev 2002;2:146-89.

Drapak Iryna Syntesis. Diuretic activity research and QSAR-analysis of N-(1,3,4-thiadiazol-2-il)substituted amides of alkanecarboxylic acids. Farmatsevtychnyi zhurnal 2019;2:80-93.

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

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.

Liliya Logoyda. Bioanalytical method development and validation from the simultaneous determination of verapamil and enalapril in the present of enalaprilat by HPLC MS/MS. Int J Appl Pharm 2018;10:19-27.

Liliya Logoyda. A HPLC-MS/MS method development and validation for the simultaneous determination of nifedipine and enalapril in human plasma. Int J Appl Pharm 2018;10:35-42.

Ghosh C, Jain I, Shinde CP, Chakraborty BS. 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 Anal 2012;4:94–103.

Bhardwaj SP, Singh S. Study of forced degradation behavior of enalapril maleate by LC and LC–MS and development of a validated stability-indicating assay method. J Pharm Biomed Anal 2008;46:113–20.

Logoyda L, Kondratova Y, Korobko D, Susla O, Soroka Y, Tsytsiura R, Pidruchna S. Youden's test of the chromatographic determination of captopril in pharmaceuticals. Int J Green Pharm 2017;11:188-91.

Liliya Logoyda, Dmytro Korobko, Oleksandra Oleshchuk, Taras Proniv, Mariya Dmutriv. A HPLC MS/MS method development and validation for the simultaneous determination of bisoprolol and enalapril in the present of enalaprilat in human plasma. Int J Appl Pharm 2018;10:31-40.