Top

Published in:

Open Access 01-07-2016 | Original Article

Pharmacokinetics of heroin and its metabolites in vitreous humor and blood in a living pig model

Authors: André Gottås, Marianne Arnestad, Per Steinar Halvorsen, Liliana C. Bachs, Gudrun Høiseth

Published in: Forensic Toxicology | Issue 2/2016

Abstract

Vitreous humor (VH) is an alternative matrix for drug analysis in forensic toxicology. However, little is known about the distribution of xenobiotics, such as opioids, into VH in living organisms. The aim of this study was to simultaneously measure heroin and metabolite concentrations in blood and VH after injection of heroin in a living pig model. Six pigs were under non-opioid anesthesia during the surgical operation and experiment. Ocular microdialysis was used to acquire dialysate from VH, and a venous catheter was used for blood sampling. Twenty milligrams of heroin was injected intravenously with subsequent sampling of blood and dialysate for 6 h. The samples were analyzed by ultra-performance liquid chromatography–tandem mass spectrometry. Heroin was not detected in VH; 6-monoacetylmorphine (6-MAM) and morphine were first detected in VH after 60 min. The morphine concentration in VH thereafter increased throughout the experimental period. For 6-MAM, C _max was reached after 230 min in VH. In blood, 6-MAM reached C _max after 0.5 min, with a subsequent biphasic elimination phase. The blood and VH 6-MAM concentrations reached equilibrium after 2 h. In blood, morphine reached C _max after 4.3 min, with a subsequent slower elimination than 6-MAM. The blood and VH morphine concentrations were in equilibrium about 6 h after injection of heroin. In conclusion, both 6-MAM and morphine showed slow transport into VH; detection of 6-MAM in VH did not necessarily reflect a recent intake of heroin. Because postmortem changes are expected to be small in VH, these experimental results could assist the interpretation of heroin deaths.

Gianutsos G, Cohen SD, Carlson G, Heyman R, Salva P, Morrow G, Hite GJ (1986) Alteration of in vivo and in vitro effects of heroin by esterase inhibition. Toxicol Appl Pharmacol 82:14–18CrossRefPubMed

Inturrisi CE, Schultz M, Shin S, Umans JG, Angel L, Simon EJ (1983) Evidence from opiate binding studies that heroin acts through its metabolites. Life Sci 33(Suppl 1):773–776CrossRefPubMed

Selley DE, Cao CC, Sexton T, Schwegel JA, Martin TJ, Childers SR (2001) µ Opioid receptor-mediated G-protein activation by heroin metabolites: evidence for greater efficacy of 6-monoacetylmorphine compared with morphine. Biochem Pharmacol 62:447–455CrossRefPubMed

Salmon AY, Goren Z, Avissar Y, Soreq H (1999) Human erythrocyte but not brain acetylcholinesterase hydrolyses heroin to morphine. Clin Exp Pharmacol Physiol 26:596–600CrossRefPubMed

Maurer HH, Sauer C, Theobald DS (2006) Toxicokinetics of drugs of abuse: current knowledge of the isoenzymes involved in the human metabolism of tetrahydrocannabinol, cocaine, heroin, morphine, and codeine. Ther Drug Monit 28:447–453CrossRefPubMed

Rook EJ, Huitema AD, van den Brink W, van Ree JM, Beijnen JH (2006) Pharmacokinetics and pharmacokinetic variability of heroin and its metabolites: review of the literature. Curr Clin Pharmacol 1:109–118CrossRefPubMed

Rook EJ, van Ree JM, van den Brink W, Hillebrand MJ, Huitema AD, Hendriks VM, Beijnen JH (2006) Pharmacokinetics and pharmacodynamics of high doses of pharmaceutically prepared heroin, by intravenous or by inhalation route in opioid-dependent patients. Basic Clin Pharmacol Toxicol 98:86–96CrossRefPubMed

Andersen JM, Ripel A, Boix F, Normann PT, Morland J (2009) Increased locomotor activity induced by heroin in mice: pharmacokinetic demonstration of heroin acting as a pro-drug for the mediator, 6-monoacetylmorphine, in vivo. J Pharmacol Exp Ther 331:153–161CrossRefPubMed

Gottås A, Øiestad EL, Boix F, Vindenes V, Ripel Å, Thaulow CH, Mørland J (2013) Levels of heroin and its metabolites in blood and brain extracellular fluid after i.v. heroin administration to freely moving rats. Br J Pharmacol 170:546–556CrossRefPubMedPubMedCentral

10.

Gottås A, Boix F, Øiestad EL, Vindenes V, Mørland J (2014) Role of 6-monoacetylmorphine in the acute release of striatal dopamine induced by intravenous heroin. Int J Neuropsychopharmacol 17:1357–1365CrossRefPubMed

11.

Gottas A, Oiestad EL, Boix F, Ripel A, Thaulow CH, Pettersen BS, Vindenes V, Morland J (2012) Simultaneous measurement of heroin and its metabolites in brain extracellular fluid by microdialysis and ultra performance liquid chromatography tandem mass spectrometry. J Pharmacol Toxicol Methods 66:14–21CrossRefPubMed

12.

Bogusz MJ, Maier RD, Driessen S (1997) Morphine, morphine-3-glucuronide, morphine-6-glucuronide, and 6-monoacetylmorphine determined by means of atmospheric pressure chemical ionization-mass spectrometry-liquid chromatography in body fluids of heroin victims. J Anal Toxicol 21:346–355CrossRefPubMed

13.

Wyman J, Bultman S (2004) Postmortem distribution of heroin metabolites in femoral blood, liver, cerebrospinal fluid, and vitreous humor. J Anal Toxicol 28:260–263CrossRefPubMed

14.

Fernandez P, Lopez-Rivadulla M, Linares JM, Tato F, Bermejo AM (1989) A comparative pharmacokinetic study of ethanol in the blood, vitreous humour and aqueous humour of rabbits. Forensic Sci Int 41:61–65CrossRefPubMed

15.

Teixeira HM, Reis F, Proença P, Ramos P, Quintela O, López-Rivadulla M, Marques E, Vieira DN (2004) Vitreous humour as a complementary sample to blood for the detection/confirmation of diazepam: ante-mortem and post-mortem studies in an animal model. Hum Exp Toxicol 23:571–577CrossRefPubMed

16.

De Letter EA, De Paepe P, Clauwaert KM, Belpaire FM, Lambert WE, Van Bocxlaer JF, Piette MH (2000) Is vitreous humour useful for the interpretation of 3,4-methylenedioxymethamphetamine (MDMA) blood levels? Experimental approach with rabbits. Int J Legal Med 114:29–35CrossRefPubMed

17.

Crandall CS, Kerrigan S, Aguero RL, Lavalley J, McKinney PE (2006) The influence of collection site and methods on postmortem morphine concentrations in a porcine model. J Anal Toxicol 30:651–658CrossRefPubMed

18.

Waga J, Ohta A, Ehinger B (1991) Intraocular microdialysis with permanently implanted probes in rabbit. Acta Ophthalmol (Copenh) 69:618–624CrossRef

19.

Waga J, Nilsson-Ehle I, Ljungberg B, Skarin A, Stahle L, Ehinger B (1999) Microdialysis for pharmacokinetic studies of ceftazidime in rabbit vitreous. J Ocul Pharmacol Ther 15:455–463CrossRefPubMed

20.

Macha S, Mitra AK (2001) Ocular pharmacokinetics of cephalosporins using microdialysis. J Ocul Pharmacol Ther 17:485–498CrossRefPubMed

21.

Katragadda S, Gunda S, Hariharan S, Mitra AK (2008) Ocular pharmacokinetics of acyclovir amino acid ester prodrugs in the anterior chamber: evaluation of their utility in treating ocular HSV infections. Int J Pharm 359:15–24CrossRefPubMedPubMedCentral

22.

Boddu SH, Gunda S, Earla R, Mitra AK (2010) Ocular microdialysis: a continuous sampling technique to study pharmacokinetics and pharmacodynamics in the eye. Bioanalysis 2:487–507CrossRefPubMed

23.

Forster R, Bode G, Ellegaard L, van der Laan JW (2010) The RETHINK project on minipigs in the toxicity testing of new medicines and chemicals: conclusions and recommendations. J Pharmacol Toxicol Methods 62:236–242CrossRefPubMed

24.

Higashi E, Ando A, Iwano S, Murayama N, Yamazaki H, Miyamoto Y (2014) Hepatic microsomal UDP-glucuronosyltransferase (UGT) activities in the microminipig. Biopharm Drug Dispos 35:313–320CrossRefPubMed

25.

Matal J, Jancova P, Siller M, Masek V, Anzenbacherova E, Anzenbacher P (2008) Interspecies comparison of the glucuronidation processes in the man, monkey, pig, dog and rat. Neuro Endocrinol Lett 29:738–743PubMed

26.

Lykkegaard K, Lykkesfeldt J, Lauritzen B, Svendsen O (2008) Morphine reduces spinal c-fos expression dose-dependently during experimental laparotomy in pigs: a combined pharmacokinetic and surgical study. Res Vet Sci 84:457–464CrossRefPubMed

27.

Karinen R, Andersen JM, Ripel A, Hasvold I, Hopen AB, Morland J, Christophersen AS (2009) Determination of heroin and its main metabolites in small sample volumes of whole blood and brain tissue by reversed-phase liquid chromatography-tandem mass spectrometry. J Anal Toxicol 33:345–350CrossRefPubMed

28.

Berg T, Lundanes E, Christophersen AS, Strand DH (2009) Determination of opiates and cocaine in urine by high pH mobile phase reversed phase UPLC-MS/MS. J Chromatogr B 877:421–432CrossRef

29.

Peters FT, Drummer OH, Musshoff F (2007) Validation of new methods. Forensic Sci Int 165:216–224CrossRefPubMed

30.

Macha S, Mitra AK (2001) Ocular pharmacokinetics in rabbits using a novel dual probe microdialysis technique. Exp Eye Res 72:289–299CrossRefPubMed

31.

Douglas W (1972) Of pigs and men and research: a review of applications and analogies of the pig, sus scrofa, in human medical research. Sp Life Sci 3:226–234

32.

Hosokawa M, Maki T, Satoh T (1990) Characterization of molecular species of liver microsomal carboxylesterases of several animal species and humans. Arch Biochem Biophys 277:219–227CrossRefPubMed

33.

Hughes PM, Olejnik O, Chang-Lin JE, Wilson CG (2005) Topical and systemic drug delivery to the posterior segments. Adv Drug Deliv Rev 57:2010–2032CrossRefPubMed

34.

Schmidt A, Ganßman B, Skopp G, Klinder K, Pötsch L, Aderjan R, Lutz R, Mattern R (1997) An in vitro experiment for postmortem vascular permeation. The passage of morphine and morphine glucuronides across a vascular wall. J Forensic Sci 42:486–491

35.

Cunha-Vaz JG (2004) The blood-retinal barriers system. Basic concepts and clinical evaluation. Exp Eye Res 78:715–721CrossRefPubMed

36.

Holmgren P, Druid H, Holmgren A, Ahlner J (2004) Stability of drugs in stored postmortem femoral blood and vitreous humor. J Forensic Sci 49:820–825CrossRefPubMed

37.

Baggot JD, Davis LE (1973) Species differences in plasma protein binding of morphine and codeine. Am J Vet Res 34:571–574PubMed

38.

Cheruvu NP, Amrite AC, Kompella UB (2008) Effect of eye pigmentation on transscleral drug delivery. Invest Ophthalmol Vis Sci 49:333–341CrossRefPubMedPubMedCentral

39.

Leblanc B, Jezequel S, Davies T, Hanton G, Taradach C (1998) Binding of drugs to eye melanin is not predictive of ocular toxicity. Regul Toxicol Pharmacol 28:124–132CrossRefPubMed

40.

Pitkanen L, Ranta VP, Moilanen H, Urtti A (2005) Permeability of retinal pigment epithelium: effects of permeant molecular weight and lipophilicity. Invest Ophthalmol Vis Sci 46:641–646CrossRefPubMed

41.

Gaudana R, Ananthula HK, Parenky A, Mitra AK (2010) Ocular drug delivery. AAPS J 12:348–360CrossRefPubMedPubMedCentral

42.

Bévalot F, Cartiser N, Bottinelli C, Fanton L, Guitton J (2016) Vitreous humor analysis for the detection of xenobiotics in forensic toxicology: a review. Forensic Toxicol 34:12–40CrossRefPubMed

43.

Kronstrand R, Forstberg-Peterson S, Kagedal B, Ahlner J, Larson G (1999) Codeine concentration in hair after oral administration is dependent on melanin content. Clin Chem 45:1485–1494PubMed

44.

Behar-Cohen FF, Gauthier S, El Aouni A, Chapon P, Parel JM, Renard G, Chauvaud D (2001) Methylprednisolone concentrations in the vitreous and the serum after pulse therapy. Retina 21:48–53CrossRefPubMed

45.

Mains J, Tan LE, Zhang T, Young L, Shi R, Wilson C (2012) Species variation in small molecule components of animal vitreous. Invest Ophthalmol Vis Sci 53:4778–4786CrossRefPubMed

46.

Drummer OH, Gerostamoulos J (2002) Postmortem drug analysis: analytical and toxicological aspects. Ther Drug Monit 24:199–209CrossRefPubMed

Title: Pharmacokinetics of heroin and its metabolites in vitreous humor and blood in a living pig model
Authors: André Gottås
Marianne Arnestad
Per Steinar Halvorsen
Liliana C. Bachs
Gudrun Høiseth
Publication date: 01-07-2016
Publisher: Springer Japan
Published in: Forensic Toxicology / Issue 2/2016
Print ISSN: 1860-8965
Electronic ISSN: 1860-8973
DOI: https://doi.org/10.1007/s11419-016-0315-z

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Pharmacokinetics of heroin and its metabolites in vitreous humor and blood in a living pig model

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 2/2016

Regioisomeric differentiation of the alkyl-substituted synthetic cannabinoids JWH-122 and JWH-210 by GC-EI-MS/MS

Application of high-resolution mass spectrometry to determination of baclofen in a case of fatal intoxication

Chiral enantioresolution of cathinone derivatives present in “legal highs”, and enantioselectivity evaluation on cytotoxicity of 3,4-methylenedioxypyrovalerone (MDPV)

Postmortem blood concentrations of sartans measured by liquid chromatography-tandem mass spectrometry

Old opioids, new concerns: the case of acetylfentanyl

Determination of 1-phenyl-2-propanone (P2P) by HS-GC/MS in a material sold as “wet amphetamine”