Simple and fast Development and validation of High Performance Liquid Chromatography Method with UV determination of thiopental in rat plasma

Authors

  • Saeed rezaee Department of pharmaceutics, school of pharmacy, Zanjan University of Medical Science, Zanjan, Iran
  • Ali asghar hemmati Department of pharmacology, school of pharmacy, Ahvaz jundishapur University of Medical Science, Khozestan, Ahvaz, Iran
  • Alireza Malayeri Nab’a Al-Hayat health research center, Nab’a Al-Hayat Foundation for Medical Sciences and Health Care, Najaf, Iraq
  • Nima Bakhtiari Medicinal planet research center, Ahvaz jundishapur University of Medical Science, Khozestan, Ahvaz, Iran

Keywords:

Chromatography, Liquid, Thiopental, Plasma, Rats

Abstract

Objective: Now days old agent thiopental return to hot-topic of research as new Model of neuropharmacology study. Because of this reason simple, fast and cost benefit method of determination this agent comes an issue in area pharmacokinetic and pharmacodynamics (PK-PD) area of research. Previous study was focus on human plasma and pilot study showed there is differentiation between human and rat plasma.

Methods: Separation were perform Nuleodur C18 ec HPLC column (250×4.0×5µm), using a mixture of acetonitrile; potassium; dihydrogen phosphate buffer (10mM, PH 2.7) as mobile phase delivered at flow rate 1.2 ml/minute. 280 nm and room temperature was selected for detection of thiopental and 1-Naphtylamine as internal standard. Plasma sample (100µl) were treated with 180µl precipitation solution and 20µl of internal standard. After the mixture were vortex and centrifuge at 1000 g, finally 40µl of clear supernatant was directly injected into 20µl loop of HPLC apparatus. Calibration curve were fitted by Peak area ratio of thiopental to internal standard. All stage of blood collection was under supervision of Ahvaz ethical committee.

Results: Thiopental and internal standard retention time were at 8.3 and 14.5 minute, respectively. 0.5 µg/ml was level of limit of quantification, of our method. 99.4 to 100.3 was range of Accuracy of methods. inter- and intraday precisions were 4-19 % and 6-8%, respectively. A good relationship in the form of a power was found (r2=0.999).

Conclusions: the presented simple fast and cost benefit method is great accurate, precise and sensitive for determination of thiopental in plasma of rat.

References

1. Russo H, Bressolle F. Pharmacodynamics and pharmacokinetics of thiopental. Clinical pharmacokinetics. 1998;35(2):95-134.
2. Lundy J, Tovell R. Some of the newer local and general anesthetic agents. Methods of their administration. Northwest Med (Seattle). 1934;33:308-11.
3. Steeds CE. The anatomy and physiology of pain. Surgery (Oxford). 2009;27(12):507-11.
4. Council NR. Guide for the care and use of laboratory animals: National Academies Press; 2010.
5. Christensen JH, Andreasen F. Determination of thiopental by high pressure liquid chromatography. Basic & Clinical Pharmacology & Toxicology. 1979;44(4):260-3.
6. Meier P, Thormann W. Determination of thiopental in human serum and plasma by high-performance capillary electrophoresis—micellar electrokinetic chromatography. Journal of Chromatography A. 1991;559(1-2):505-13.
7. Shiu GK, Nemoto EM. Simple, rapid and sensitive reversed-phase high-performance liquid chromatographic method for thiopental and pentobarbital determination in plasma and brain tissue. Journal of Chromatography B: Biomedical Sciences and Applications. 1982;227(1):207-12.
8. Sennello LT, Kohn FE. Gas chromatographic determination of thiopental in plasma using an alkali flame ionization detector. Analytical chemistry. 1974;46(6):752-5.
9. Carroll F, Smith D, Mark L, Brand L, Perel J. Determination of optically active thiopental, thiamylal, and their metabolites in human urine. Drug Metabolism and Disposition. 1977;5(4):343-54.
10. Becker KE. Plasma levels of thiopental necessary for anesthesia. Anesthesiology. 1978;49(3):192-6.
11. Jung D, Mayersohn M, Perrier D. Gas-chromatographic assay for thiopental in plasma, with use of a nitrogen-specific detector. Clinical chemistry. 1981;27(1):113-5.
12. Gustafsson LL, Ebling WF, Osaki E, Stanski DR. Quantitation of depth of thiopental anesthesia in the rat. Anesthesiology: The Journal of the American Society of Anesthesiologists. 1996;84(2):415-27.
13. Braddock LI, Marec N. The gas chromatographic analysis of sub-microgram quantities of barbiturates using a flame ionization detector. Journal of Chromatographic Science. 1965;3(8):274-7.
14. Schepens P, Heyndrickx A. Placental transfer of thiopental. Eur J Toxicol. 1975;8(2):87-93.
15. Becker Jr K. Gas chromatographic assay for free and total plasma levels of thiopental. Anesthesiology. 1976;45(6):656-60.
16. Stanski DR, Burch PG, Harapat S, Richards RK. Pharmacokinetics and anesthetic potency of a thiopental isomer. Journal of pharmaceutical sciences. 1983;72(8):937-40.
17. Burch PG, Stanski DR. Decreased protein binding and thiopental kinetics. Clinical Pharmacology & Therapeutics. 1982;32(2):212-7.
18. Christensen JH, Andreasen F. Individual variation in response to thiopental. Acta Anaesthesiologica Scandinavica. 1978;22(3):303-13.
19. Burch PG, Stanski DR. The role of metabolism and protein binding in thiopental anesthesia. Anesthesiology. 1983;58(2):146-52.
20. Polson C, Sarkar P, Incledon B, Raguvaran V, Grant R. Optimization of protein precipitation based upon effectiveness of protein removal and ionization effect in liquid chromatography–tandem mass spectrometry. Journal of Chromatography B. 2003;785(2):263-75.

Downloads

Published

2018-06-26

How to Cite

rezaee, S., hemmati, A. asghar, Malayeri, A., & Bakhtiari, N. (2018). Simple and fast Development and validation of High Performance Liquid Chromatography Method with UV determination of thiopental in rat plasma. Journal of Contemporary Medical Sciences, 4(2), 78–81. Retrieved from https://www.jocms.org/index.php/jcms/article/view/403

Issue

Section

Articles

Most read articles by the same author(s)