Email this article A simple and sensitive HPLC method for analysis of imipramine in human plasma with UV detection and liquid-liquid extraction: Application in bioequivalence studies
Abstract
High-performance liquid chromatography (HPLC) methods employing ultraviolet (UV) detector are not sufficiently sensitive to measure the low plasma concentrations following single oral dose of imipramine. Therefore, in the present study a simple, rapid and yet sensitive HPLC method with UV detection was developed and validated for quantitation of imipramine in human plasma samples. An efficient liquid-liquid extraction (LLE) of imipramine from plasma with the mixture of hexane/isoamyl alcohol (98:2) and back extraction of the drug in acidic medium concomitant with evaporation of organic phase allowed the use of UV detector to conveniently measure plasma levels of this compound as low level as 3 ng/ml. Separation was achieved on a µ-Bondapak C18 HPLC column using sodium hydrogen phosphate solution (0.01 M)/acetonitrile (60/40 v/v) at pH 3.5 ± 0.1 at 1.5 ml/min. Trimipramine was used as the internal standard for analysis of plasma samples. The retention times for imipramine and trimipramine were 4.3 and 5.2 min, respectively. Calibration curve was linear in the range of 3-40 ng/ml using human plasma with the average extraction recovery of 85 ± 5%. Imipramine was found to be stable in plasma samples with no evidence of degradation during three freeze-thaw cycles and three months storage at -70 ºC. The current validated method was finally applied in bioequivalence studies of two different imipramine products according to a standard two-way crossover design with a two weeks washout period.
Full Text:
PDFReferences
Gillman PK. Tricyclic antidepressant pharmacology and therapeutic drug interactions updated. Br J Pharmacol. 2007;151:737–748.
Klerman GL,J.O Coli.. Clinical pharmacology of imipramine and related antidepressant compounds. Pharmacological Reviews. 1965:17:101-141.
Patel SK, Patel NJ. Simultaneous determination of imipramine hydrochloride and chlordiazepoxide in pharmaceutical preparations by spectrophotometric, RP-HPLC, and HPTLC methods. J AOAC Int. 2010;93:904-910.
Weigmann H, Hartter S, Bagli M, Hiemke C. Steady state concentrations of clomipramine and its major metabolite desmethylclomipramine in rat brain and serum after oral administration of clomipramine. Eur Neuropsychopharmacol. 2000;10:401–405.
Kobayashi A, Sugita S, Nakazawa K. High-performance liquid chromatographic determination of imipramine and desipramine in human serum. J Chromatogr. 1984;336:410-440.
Samanidou VF, Nika MK, Papadoyannis IN. Development of an HPLC method for the monitoring of tricyclic antidepressants in biofluids. J Sep Sci. 2007;30:2391-2400.
Mohamed FA, Mohamed HA, Hussein SA, Ahmed SA. A validated spectrofluorimetric method for determination of some psychoactive drugs. J Pharmaceutical Biomed Anal. 2005;39:139–146.
Hammer MW, Brodiet BB. Application of isotope derivative technique to assay of secondary amines: estimation of desipramine dy acetylation with H3-acetic anhydride. J Pharmacol Exp Ther. 1967;157:503-508.
Frigerio A, Belvedere G, De Nadai F, Fanelli R, Pantarotto C, Riva E, et al. A method for the determination of imipramine in human plasma by gas-liquid chromatography-mass fragmentography. J Chromatogr A. 1972;74:201-208.
Nielsen KK, Brøsen K. High-performance liquid chromatography of imipramine and six metabolites in human plasma and urine. J Chromatogr. 1993;1:87-94.
Önal A, Öztunç A. A rapid and simple RP-HPLC method for quantification of desipramine in human plasma. Reviews in Analytical Chemistry. 2011;30:165–169.
Stout SA, DeVane CL. Quantification of imipramine and its major metabolites in whole blood, brain, and other tissues of the rat by liquid chromatography. Psychopharmacology. 1984;84:39-41.
Messiha FS. Determination of carbamazepine by HPLC electrochemical detection and application for estimation of imipramine desipramine, doxepin and nordoxepin. Alcohol. 1986;3:135-138.
Chen AG, Wing YK, Chiu H, Lee S, Chen CN, Chana K. Simultaneous determination of imipramine, desipramine and their 2- and 10-hydroxylated metabolites in human plasma and urine by high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl. 1997;693:153–158.
Thongnopnua P, Karnjanaves K. The rapid simultaneous tricyclic antidepressant determination by plasma deproteinization and liquid chromatography. Asian Biomed. 2008;2:305-318.
Vemula VRB, Sharma PK. Analytical method development and validation for simultaneous estimation of imipramine and diazepam in tablet dosage form by RP-HPLC. Int J Pharm Pharm Sci. 2013:5:249-253.
Aymard G, Livi P, Pham YT, Diquet B. Sensitive and rapid method for the simultaneous quantification of five antidepressants with their respective metabolites in plasma using high-performance liquid chromatography with diode-array detection. J Chromatogr B Biomed Sci Appl. 1997;700:183-189.
Titier K, Castaing N, Le-Déodic M, Le-Bars D, Moore N, Molimard M. Quantification of tricyclic antidepressants and monoamine oxidase inhibitors by high performance liquid chromatography-tandem mass spectrometry in whole blood. J Anal Toxicol. 2007;31:200-207.
Nasir M, Ioannis S, Papadoyannis N. Development and validation of an HPLC method for the determination of benzodiazepines and tricyclic antidepressants in biological fluids after sequential SPE. J Sep Sci. 2008;31:2358–2370.
Emami J, Rezazadeh M, Kalani M. Quantification of pantoprazole by high performance liquid chromatography (HPLC) method: In vitro and in vivo application. J Liq Chrom Relat Technol. 2014;37:681-695.
Emami J, Ghassami N, Ahmadi F. Development and validation of a new HPLC method for determination of lamotrigine and related compounds in tablet formulations. J Pharm Biomed Anal. 2006;40: 999-1005.
Emami J, Rezazadeh M. Rapid, sensitive and validated HPLC method for analysis of metronidazole and tinidazole under identical chromatographic conditions with UV detection and liquid-liquid extraction: application in bioequivalence studies. Actachromatographica. 2013;25:111-125.
Deepakumari H N, Vinay KB, Revanasiddappa HD. Development and validation of a stability indicating RP-UPLC method for analysis of imipramine hydrochloride in pharmaceuticals. Int Scholarly Research Notices. 2013;2013:ID 913765.
Bose D, Martinavarro-Domínguez A, Gil-Agustí M, Carda-Broch S, Durgbanshi A, Capella-Peiró M, et al. Therapeutic monitoring of imipramine and desipramine by micellar liquid chromatography with direct injection and electrochemical detection. Biomed Chromatogr. 2005;19:343–349.
Refbacks
- There are currently no refbacks.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.