QUANTIFICATION OF URAPIDIL IN HUMAN PLASMA USING ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY–ELECTROSPRAY IONIZATION MASS SPECTROMETRY (UPLC–MS/MS) FOR PHARMACOKINETIC STUDY IN HEALTHY INDIAN VOLUNTEERS

  • Ashish Saxena Bioanalytical Research Department, Lupin Bio-Research Center, Pashan, Pune 411021, Maharastra State, India. *PhD Research scholar, Faculty of Pharmacy, Pacific University, P.B.-12 Pacific Hills, Airport Road, Pratap Nagar Extension, Debari, Udaipur 313024, Rajasthan State, India
  • Arun Kumar Gupta Pacific University
  • Praveen Kumar V Lupin Bio-Research Center Pacific University
  • M. Sundaramoorthi Nainar Lupin Bio-Research Center
  • Manoj Bob Lupin Bio-Research Center
  • Ravisekhar Kasibhatta Lupin Bio-Research Center

Abstract

Objective: A rapid and selective quantitative method was developed and validated in human plasma for urapidil pharmacokinetic study in healthy Indian volunteers.

Methods: The ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) method with solid-phase extraction technique utilized Strata X 33µ polymeric reversed phase (30 mg/mL), extraction cartridge. Simple gradient chromatographic conditions and selective reaction monitoring in mass spectrometric detection enabled accurate and precise measurement of urapidil at nanogram levels in 0.1 mL of human plasma. The method used a deuterium labeled internal standard.

Results: The method was validated for a linear range of 5–500 ng/mL for urapidil with a correlation coefficient ³ 0.99 The intra-run and inter-run precision and accuracy were within 10%. The overall recoveries for urapidil and urapidil D4 were more than 90%. The urapidil was found to be stable in plasma matrix and aqueous media.

Conclusion: The developed and validated method was specific, sensitive and reproducible in the analysis of clinical samples interspersed with quality control samples under freshly prepared calibration standards. The method was applied for the determination of the pharmacokinetic parameters of urapidil following a single oral administration of urapidil 60 mg capsules in nineteen healthy Indian male volunteers for fasting and fed study.

 

Keywords: Urapidil, UPLC–MS/MS, Human plasma, Pharmacokinetic study, Solid phase extraction

Downloads

Download data is not yet available.

Author Biography

Ashish Saxena, Bioanalytical Research Department, Lupin Bio-Research Center, Pashan, Pune 411021, Maharastra State, India. *PhD Research scholar, Faculty of Pharmacy, Pacific University, P.B.-12 Pacific Hills, Airport Road, Pratap Nagar Extension, Debari, Udaipur 313024, Rajasthan State, India

PhD Research scholar, Faculty of Pharmacy

References

1. Schoetensack W, Bruckschen EG, Zech K. Urapidil, in new drugs annual: cardiovascular drugs. Cardiovasc Drug Rev 1983;1:19–48.
2. Ramage AG. Influence of 5-HT1A receptor agonists on sympathetic and parasympathetic nerve activity. J Cardiovasc Pharmacol 1990;15 Suppl 7:S75-85.
3. Kolassa N, Beller KD, Sanders KH. Involvement of brain 5-HT1A receptors in the hypotensive response to urapidil. Am J Cardiol 1989;64(7):7D–10D.
4. Sanders KH, Beller KD, Bischler P, Kolassa N. Interactions of urapidil with brain serotonin-1A receptors increases the blood pressure reduction due to peripheral alpha-adrenoceptor inhibition. J Hypertens 1988;6(2):S65–S8.
5. Mandal AK, Kellar KJ, Friedman E, Pineo SV, Hamosh P, Gillis RA. Importance of central nervous system serotonin-1A receptors for mediating the hypotensive effects of urapidil. J Pharmacol Exp Ther 1989;251(2):563–70.
6. Shebuski RJ, Zimmerman BG. Suppression of reflex tachycardia following alpha-adrenoceptor blockade in conscious dogs: Comparison of urapidil with prazosin. J Cardiovasc Pharmacol 1984;6(5):788–94.
7. Hanft G, Gross G. Subclassification of alpha 1-adrenoceptor recognition sites by urapidil derivatives and other selective antagonists. Br J Pharmacol 1989;97(3):691–700.
8. Haenni A, Lithell H. Urapidil treatment decreases plasma fibrinogen concentration in essential hypertension. Metab 1996;45(10):1221–29.
9. Zech K, Eltze M, Kilian U, Sanders KH, Kolassa N. Biotransformation of urapidil: metabolites in serum and urine and their biological activity in vitro and in vivo. Biomed Chromatogr 2011;25(12):1319–26.
10. Ramage AG. The mechanism of the sympathoinhibitory action of urapidil: role of 5-HT1A receptors. Br J Pharmacol 1991;102(4):998–1002.
11. Eltze M. Investigations on the mode of action of a new antihypertensive drug, urapidil, in the isolated rat vas deferens. Eur J Pharmacol 1979;59(1-2):1–9.
12. Verberne AJ, Rand MJ. Pharmacological activities of the antihypertensive drug urapidil in the rat. Clin Exp Pharmacol Physiol 1984;11(4):407–11.
13. Verberne AJ, Rand MJ. Effect of urapidil on beta-adrenoceptors of rat atria. Eur J Pharmacol 1985;108(2):193–6.
14. Bottorff MB, Hoon TJ, Rodman JH, Gerlach PA, Ramanathan KB. Pharmacokinetics and pharmacodynamics of urapidil in severe hypertension. J Clin Pharmacol 1988;28(5):420–6.
15. Kirsten R, Nelson K, Molz KH, Haerlin R, Steinijans VW. Pharmacodynamics and pharmacokinetics of urapidil in hypertensive patients: a crossover study comparing infusion with an infusion-capsule combination. Eur J Clin Pharmacol 1987;32(1):61–5.
16. Hansson L. Treatment of hypertension in the elderly with special reference to urapidil. Blood Press Suppl 1994;4:45–8.
17. Veltkamp AC, Das HA, Frei RW, Brinkman UA. On-line radiometric determination of [14C]-urapidil and its main metabolises in rat plasma, using post-column ion-pair extraction and solvent segmentation techniques. J Pharm Biomed Anal 1988;6:609–22.
18. Zech K, Huber R. Determination of urapidil and its metabolites in human serum and urine: comparison of liquid-liquid and fully automated liquid-solid extraction. J Chromatogr 1986;353:351–60.
19. Yue Q, Song Z, Wang CJ. Rapid determination of subnanogram urapidil using flow Injection Enhancement Chemiluminesence. J Anal Chem 2006;61:295–9.
20. Nirogi R, Vishwottam K, Prashanth K, Raghupathi A, Rajesh B, Pavan KM. Quantification of urapidil, a-1-adrenoreceptor antagonist, in plasma by LC-MS/MS: validation and application to pharmacokinetic studies. Biomed Chromatogr 2010;25:1319–26.
21. Ambavaram VBR, Nandigam V, Vemula M, Kalluru GR, Gajulapalle M. Liquid chromatography-tandem mass spectrometry method for simultaneous quantification of urapidil and aripiprazole in human plasma and its application to human pharmacokinetic study. Biomed Chromatogr 2013;27(7):916–23.
22. Guidance for Industry, Bioanalytical Method Validation, US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for Biologics Evaluation and Research (CBER); 2001.
23. Guideline on Bioanalytical Method Validation. EMEA/CHMP/EWP/192217/2009, Committee for Medicinal Products for Human Use (CHMP); 2011.
24. Almeida AM, Castel-Branco MM, Falcao AC. Linear regression for calibration lines revisited: weighting schemes for bioanalytical methods. J Chromatogr B 2002;774:215–22.
Statistics
263 Views | 836 Downloads
How to Cite
Saxena, A., A. K. Gupta, P. K. V, M. S. Nainar, M. Bob, and R. Kasibhatta. “QUANTIFICATION OF URAPIDIL IN HUMAN PLASMA USING ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY–ELECTROSPRAY IONIZATION MASS SPECTROMETRY (UPLC–MS/MS) FOR PHARMACOKINETIC STUDY IN HEALTHY INDIAN VOLUNTEERS”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 6, no. 10, 1, pp. 565-70, https://innovareacademics.in/journals/index.php/ijpps/article/view/2902.
Section
Original Article(s)