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Postmortem Detection of Olanzapine and Clozapine Residues in Rat Brain following different Modes of Death using HPLC-UV Technique

Author Affiliations

  • 1Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Zagazig University, Egypt
  • 2Chemistry Department, Faculty of Science, Zagazig University, Egypt
  • 3Chemical Laboratory, forensic medicine authority, ministry of justice, Egypt

Res. J. Forensic Sci., Volume 4, Issue (7), Pages 1-15, August,29 (2016)


The aim of this study was directed to develop and validate an HPLC with UV detection method for quantification of Olanzapine (OLZ) and Clozapine (CLZ) in rat brain tissue homogenate following different modes of death, for this purpose sixty adult male albino rats were classified into four equal groups; first group (control) received distilled water, second group received OLZ at a dose of 5 mg/kg B.wt, third group received CLZ at a dose of 20 mg/kg B.wt. and fourth group received both OLZ and CLZ at the same previously mentioned doses. The administration was carried out daily by gavage for four months. At the end of the experiment; the four main groups were classified according to mode of death into three equal subgroups; drowning group, electrocution group and starvation group. The analytes and risperidone (IS) were isolated from rat brain tissue homogenate by solid phase extraction on Waters Oasis HLB cartridges. The compound was separated on a kinetex C18 column (100×4.6 mm, 2.6 µm) using a mixture of acetonitrile (A), methanol (B) and 34 mM phosphate buffer pH 2.4 (C) as mobile phase at a flow rate of 1.0 mL/min while detection wavelength was 254 nm. Under these conditions, OLZ and CLZ were well separated and showed good linearities in the ranges of 0.5–100 ng/g for OLZ and 0.5–200 ng/g for CLZ with r &


  1. Roman M., Kronstrand R., lindstedt D. and Josefsson M. (2008)., Quantitation of Seven Low-Dosage Antipsychotic Drugs in Human Postmortem Blood Using LC-MS-MS., J. Anal. Toxicol., 32(2), 147-155.
  2. Kapur S. and Remington G. (2001)., Atypical antipsychotics: new directions and new challenges in the treatment of schizophrenia., Annu. Rev. Med., 52, 503-517.
  3. Fulton B. and Goa K.L. (1997)., Olanzapine: a review of its pharmacological properties and therapeutic efficacy in the management of schizophrenia and related psychoses., Drugs, 53(2), 281-298.
  4. McConnell J., Kirby R. and Rudloff E. (2007)., Administration of acepromazine meleate to 31 dogs with history of seizures., J. Vet. Emerg. Crit. Care, 17(3), 262-267.
  5. Halici Z., Keles O.N., Unal D., Albayrak M., Suleyman H., Cadirci E., Unal B. and Kaplan S. (2007)., Chronically Administered Risperidone Did Not Change the Number of Hepatocytes in Rats: A Stereological and Histopathological Study., Basic. Clin. Pharmacol. Toxicol., 102(5), 426-432.
  6. Jafari S., Bouillon M.E., Huang X.F., Pyne S.G. and Fernandez-Enright F. (2012)., Novel olanzapine analogues presenting a reduced H1 receptor affinity and retained 5HT2A/D2 binding affinity ratio., J. Neurochem., 120, 371-384.
  7. Nandra K.S. and Agius M. (2012)., The differences between typical and atypical antipsychotics: The effects on neurogenesis., Psychiatr. Danub., 24 , 95-99.
  8. Frangou S. and Lewis M. (2000)., Atypical antipsychotics in ordinary clinical practice: a pharmaco-epidemiologic survey in a south London service., Eur. Psychiatr., 15(3), 220-226.
  9. Kollroser M. and Schober C. (2002)., Direct-injection high performance liquid chromatography ion trap mass spectrometry for the quantitative determination of olanzapine, clozapine and N-desmethylclozapine in human plasma., Rapid Commun. Mass Spectrom., 16(13), 1266-1272.
  10. Sanchez C., Martinez M. A. and Almarza E. (2005)., Determination of several psychiatric drugs in whole blood using capillary gas–liquid chromatography with nitrogen phosphorous detection comparison of two solid phase extraction procedures., Forensic Sci. Int., 155(2-3), 193-204.
  11. Markowitz J.S. and Patrick K.S. (1995)., Thermal degradation of clozapine-N-oxide to clozapine during gas chromatographic analysis., J. Chromatogr. B., 668, 171-117.
  12. Ulrich S. (2005)., Assay of olanzapine in human plasma by a rapid and sensitive gas chromatography-nitrogen phosphorus selective detection (GC-NPD) method: validation and comparison with high-performance liquid chromatography-coulometric detection., Ther. Drug Monit., 27(4), 463-468.
  13. Zhang G., Terry J.V. and Bartlett M.G. (2007)., Simultaneous determination of five antipsychotic drugs in rat plasma by high performance liquid chromatography with ultraviolet detection., J. Chromatogr. B., 856(1-2), 20–8.
  14. Garcia L.G., Bears. I.F., Pehourcq F. and Jarry C. (2003)., Simultaneous determination of four antipsychotic drugs in plasma by high performance liquid chromatography application to management of acute intoxications., J. Chromatogr. B., 795(2), 257-264.
  15. Wong J.O., Leung S., Mak T., Chan K., Cheung H.H., Choi W., Lai J. and Tsang A.W. (2006)., Plasma clozapine levels and clinical response in treatment-refractory Chinese schizophrenic patients., Prog. Neuro-Psychoph., 30(2), 251-264.
  16. Mercolini L., Bugamelli F., Kenndler E., Boncompagni G., Franchini L. and Raggi M.A. (2007)., Simultaneous determination of the antipsychotic drugs levomepromazine and clozapine and their main metabolites in human plasma by a HPLC-UV method with solid-phase extraction., J. Chromatogr. B. Analyt Technol. Biomed. Life Sci., 846(1-2), 273-280.
  17. Mosier K.E., Song J. and McKay G.J. (2003)., Determination of clozapine, and its metabolites, N-desmethylclozapine and clozapine N-oxide in dog plasma using high-performance liquid chromatography., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 783(2), 377-382.
  18. Shen Y.L., Wu H.L., Ko. W.K. and Wu S.M. (2002)., Simultaneous determination of clozapine, clozapine N-oxide, N-desmethylclozapine, risperidone and 9-hydroxyrisperidone in plasma by high performance liquid chromatography with ultraviolet detection., J. Anal. Chim. Acta, 460(2), 201-208.
  19. Llerena A., Berecz R., Norberto M.J. and Rubia A. (2001)., Determination of clozapine and its N-desmethyl metabolite by high-performance liquid chromatography with ultraviolet detection., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 755(1-2), 349-354.
  20. Boulton W.D., Markowitz J.S. and Devane C.L. (2001)., A high-performance liquid chromatography assay with ultraviolet detection for olanzapine in human plasma and urine., J. Chromatogr. B., 759(2), 319-323.
  21. Dusci L.J., Peter H.L. and Fellows L.M. (2002)., Determination of olanzapine in plasma by high-performance liquid chromatography using ultraviolet absorbance detection., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 773(2), 191-197.
  22. Jasinska A., Starczewska B., Polkowska M. and Puzanowska T.H. (2005)., Solid phase extraction of olanzapine with reverse phase sorbents prior to UV and HPLC analysis., Anal. Lett., 38(5), 815-824.
  23. D, Determination of olanzapine inhuman plasma by reversed-phase high-performance liquid chromatography with ultraviolet detection., Ther. Drug Monit., 28(3), 388-393.
  24. Sabbioni C., Saracino M.A., Mandrioli R., Albers L., Boncompagni G. and Raggi M.A. (2004)., Rapid analysis of olanzapine and desmethylolanzapine in human plasma using high-performance liquid chromatography with coulometric detection., Anal. Chim. acta, 516(1-2), 111-117.
  25. Suckow R.F., Fein M., Correll C.V. and Cooper T.B. (2004)., Determination of plasma ziprasidone using liquid chromatography with fluorescence detection., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 799 (2), 201-208.
  26. Bao J. and Potts B.D. (2001)., Quantitative determination of olanzapine in rat brain tissue by high-performance liquid chromatography with electrochemical detection., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 752(1), 61-67.
  27. Saracino M. A., Gandolfi O., Albars L., Kenndler E. and Raggi M.A. (2006)., Determination of olanzapine in rat brain using high-performance liquid chromatography with coulometric detection and a rapid solid-phase extraction procedure., J. Chromatogr. A., 1122(1-2), 21-27.
  28. Titier K., Bouchet S., Pehourcq F., Moore N. and Molimard M. (2003)., High-performance liquid chromatographic method with diode array detection to identify and quantify atypical antipsychotics and haloperidol in plasma after overdose., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 788(1), 179-185.
  29. Aravagiri M., Yuwiler A. and Marder S.R. (1998)., Distribution after repeated oral administration of different dose levels of risperidone and 9-hydroxy-risperidone in the brain and other tissues of rat., Psychopharmacology (Berl), 139(4), 356-363.
  30. Kratzsch C., Peters F.T., Kraemer T., Weber A.A. and Maurer H.H. (2003)., Screening, library-assisted identification and validated quantification of fifteen neuroleptics and three of their metabolites in plasma by liquid chromatography/mass spectrometry with atmospheric pressure chemical ionization., J. Mass Spectrom., 38(3), 283-295.
  31. Niederlander H.A., Koster E.H., Hilhorst M.J., Metting H.J., Eilders M., Ooms B. and de Jong G.J. (2006)., High throughput therapeutic drug monitoring of clozapine and metabolites in serum by on-line coupling of solid phase extraction with liquid chromatography-mass spectrometry., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 834(1-2), 98-107.
  32. Rao L.V., Snyder M.L. and Vallaro G.M. (2009)., Rapid liquid chromatography/tandem mass spectrometer (LCMS) method for clozapine and its metabolite N-desmethyl clozapine (norclozapine) in human serum., J. Clin. Lab. Anal., 23(6), 394-398.
  33. Wohlfarth A., Toepfner N., Hermanns-Clausen M. and Auwarter V. (2011)., Sensitive quantification of clozapine and its main metabolites norclozapine and clozapine-N-oxide in serum and urine using LC–MS/MS after simple liquid–liquid extraction work-up., Anal. Bioanal. Chem., 400(3), 737-746.
  34. Dinesh P.S., Naveen S.C., Mukesh P.N., Bhavin P.S., Shrivastav P. and Sanyal M. (2012)., LC–MS/MS assay for olanzapine in human plasma and its application to a bioequivalence study., Acta Pharm. Sinic. B., 2(5), 481-494.
  35. Josefsson M., Roman M., Skogh E. and Dahl M.L. (2010)., Liquid chromatography/tandem mass spectrometry for determination of olanzapine and N-33-desmethylolanzapine in human serum and cerebrospinal fluid., J. Pharm Biomed. Anal., 53(3), 576-582.
  36. Nielsen M.K. and Johansen S.S. (2009)., Determination of olanzapine in whole blood using simple protein precipitation and liquid chromatography–tandem mass spectrometry., J. Anal. Toxicol., 33(4), 15-26.
  37. Saar E., Gerostamoulos D., Drummer O.H. and Beyer J. (2010)., Identification and quantification of 30 antipsychotics in blood using LC–MS/MS., J. Mass Spectrom., 45(8), 915-925.
  38. Nielsen M.K., Johansen S.S., Dalsgaard P.W. and Linnet K. (2010)., Simultaneous screening and quantification of 52 common pharmaceuticals and drugs of abuse in hair using UPLC–TOF-MS., Forensic Sci. Int., 196(1-3), 85-92.
  39. Zhang G., Terry A.V. and Bartlett M.G. (2007)., Sensitive liquid chromatography/tandem mass spectrometry method for the simultaneous determination of olanzapine, risperidone, 9-hydroxyrisperidone, clozapine, haloperidol and ziprasidone in rat brain tissue., Rapid Commun. Mass Spectrom., 21(6), 920-928.
  40. Bhatt J., Subbaiah G. and Singh S. (2006)., Liquid chromatography/tandem mass spectrometry method for the simultaneous determination of olanzapine, risperidone, 9-hydroxyrisperidone, clozapine, haloperidol and ziprasidone in rat plasma., Rapid Commun. Mass Spectrom., 20(14), 2109-2114.
  41. Aravagiri M. and Marder S.R. (2002)., Brain, plasma and tissue pharmacokinetics of risperidone and 9-hydroxyrisperidone after separate oral administration to rats., Psychopharmacology (Berl), 159(4), 424-431.
  42. Bancroft John and Stevens Alan (1996)., Theory and practice of technique., 4th ed. New. York: Churchill Livingston, 766. ISBN: 044304760X.
  43. Castaing N., Titier K., CanaI-Raffin M., Moore N. and Molimard M. (2006)., Postmortem Redistribution of Two Antipsychotic Drugs Haloperidol and Thioridazine, in the Rat., J. Anal. Toxicol., 30(7), 419-425.
  44. Zhou Z., Li X., Li K., Xie Z., Cheng Z., Peng W., Wang F., Zhu R. and Li H. (2004)., Simultaneous determination of clozapine, olanzapine, risperidone and quetiapine in plasma by high-performance liquid chromatography–electrospray ionization mass spectrometry., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 802(2), 257-262.
  45. Tyson S.C., Devane C.L. and Risch S.C. (1995)., Pharmacokinetic interaction between risperidone and clozapine., Am. J. Psychiatry, 152, 1401-1402.
  46. Zhou S.F., Xue C.C. and Yu X.Q. (2007)., Clinically important drug interactions potentially involving mechanism-based inhibition of cytochrome p450 3A4 and the role of therapeutic drug monitoring., Ther. Drug Monit., 29, 687-710.
  47. Van Haren, N.E. Schnack, H.G. Cahn, W. van den Heuvel, M.P. Lepage, C. Collins, L. Evans, A. C. Hulshoff H.E. and Kahn R.S. (2011)., Changes in Cortical Thickness during the Course of Illness in Schizophrenia., Arch. Gen. Psychiatry, 68(9), 871-880.
  48. Vernon A.C., Natesan S., Modo M. and Kapur S. (2011)., Effect of Chronic Antipsychotic Treatment on Brain Structure: A Serial Magnetic Resonance Imaging Study with Ex Vivo and Postmortem Confirmation., Biol. Psychiatry, 69(10), 936-944.
  49. Zhao C.S., Puurunen K., Schallert T., Sivenius J. and Jolkkonen J. (2005)., Behavioral and histological effects of chronic antipsychotic and antidepressant drug treatment in aged rats with focal ischemic brain injury., Behav. Brain, Res., 158(2), 211-220.
  50. Perera J., Tan J.H., Jeevathayaparan S., Chakravarthi S. and Haleagrahara N. (2011)., Neuroprotective effects of alpha lipoic Acid on haloperidol-induced oxidative stress in the rat brain., Cell Biosci., 22(1), 2045-3701.