Nicotine e-cigarette vapor inhalation effects on nicotine & cotinine plasma levels and somatic withdrawal signs in adult male Wistar rats

  • Christian MontanariEmail author
  • Leslie K. Kelley
  • Tony M. Kerr
  • Maury Cole
  • Nicholas W. Gilpin
Original Investigation



Non-contingent chronic nicotine exposure procedures have evolved rapidly in recent years, culminating in electronic nicotine delivery systems (ENDS or e-cigarettes) to deliver vaporized drugs to rodents in standard housing chambers.


The aim of the current work was to use ENDS to test concentration-dependent effects of nicotine e-cigarette vapor inhalation on blood-nicotine concentrations, blood-cotinine concentrations, and somatic withdrawal signs over time in rats.


Male Wistar rats were exposed to vapor containing various concentrations of nicotine (20, 40, 80 mg/mL) for 11 days through ENDS, and blood concentrations of nicotine and cotinine, the major proximate metabolite of nicotine, as well as spontaneous and precipitated somatic withdrawal signs, were measured over time (across days of exposure and over hours after termination of vapor exposure).


Exposing male Wistar rats to non-contingent nicotine vapor inhalation through ENDS produces somatic withdrawal symptoms and measurable blood-nicotine and blood-cotinine levels that change according to (1) concentration of nicotine in vape solution, (2) number of days of nicotine vapor exposure, (3) time since termination of nicotine vapor exposure, and (4) relative to the withdrawal signs, whether withdrawal was spontaneous or precipitated (by mecamylamine).


The data presented here provide parameters that can be used as a reasonable starting point for future work that employs ENDS to deliver non-contingent nicotine vapor in rats, although many parameters can and should be altered to match the specific goals of future work.


Nicotine ENDS E-cigarette Vape Withdrawal Addiction 


Funding information

Funding for this award was provided by the National Institute of Drug Abuse (NIDA) Award R44DA046300 (MC and NWG). This work was supported in part by Merit Review Award no. I01 BX003451 (NWG) from the United States (U.S.) Department of Veterans Affairs, Biomedical Laboratory Research and Development Service.

Compliance with ethical standards

Conflict of interest

Nicholas W. Gilpin owns shares in Glauser Life Sciences Inc., a company with activities aimed at developing medications for treating mental health disorders; this affiliation had no direct association with the work presented here. Maury Cole is the CEO of LJARI (La Jolla Alcohol Research, Inc.), an organization dedicated to the design and manufacturing of the inhalation systems we purchased.


  1. Bacher I, Wu B, Shytle DR, George TP (2009) Mecamylamine - a nicotinic acetylcholine receptor antagonist with potential for the treatment of neuropsychiatric disorders. Expert Opin Pharmacother 10(16):2709–2721. CrossRefPubMedGoogle Scholar
  2. Baiamonte BA, Valenza M, Roltsch EA, Whitaker AM, Baynes BB, Sabino V, Gilpin NW (2014) Nicotine dependence produces hyperalgesia: role of corticotropin-releasing factor-1 receptors (CRF1Rs) in the central amygdala (CeA). Neuropharmacology 77:217–223. CrossRefPubMedGoogle Scholar
  3. Benowitz NL (1984) The use of biologic fluid samples in assessing smoke consumption. In: Measurement in the Analysis and Treatment of Smoking Behavior. NIDA Monograph 48 (Grabowski J, Bell CS, eds). Washington: U.S. Government Printing Office 1984:6–26Google Scholar
  4. Bold KW, Kong G, Camenga DR, et al (2017) Trajectories of e-cigarette and conventional cigarette use among youth. Pediatrics. 141(1). doi: Scholar
  5. Chaffee BW, Watkins SL, Glantz SA (2018) Electronic cigarette use and progression from experimentation to established smoking. Pediatrics. 141(4). doi: Scholar
  6. Cohen A, George O (2013) Animal models of nicotine exposure: relevance to second-hand smoking, electronic cigarette use, and compulsive smoking. Front Psychiatry 4:41. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Cohen A, Koob GF, George O (2012) Robust escalation of nicotine intake with extended access to nicotine self-administration and intermittent periods of abstinence. Neuropsychopharmacol 37:2153–2160. CrossRefGoogle Scholar
  8. Craig EL, Zhao B, Cui JZ, Novalen M, Miksys S, Tyndale RF (2014) Nicotine pharmacokinetics in rats is altered as a function of age, impacting the interpretation of animal model data. Drug Metab Dispos 42(9):1447–1455. CrossRefPubMedPubMedCentralGoogle Scholar
  9. U.S. Department of Health, and Human Services, Centers for Disease Control and Prevention, National Center for Chronic Disease, Prevention and Health Promotion, Office on Smoking and Health. E-cigarette use among youth and young adults: a report of the surgeon general - executive summary; 2016. Accessed July 30, 2019.
  10. Epping-Jordan MP, Watkins SS, Koob GF, Markou A (1998) Dramatic decreases in brain reward function during nicotine withdrawal. Nature 393(6680):76–79CrossRefGoogle Scholar
  11. George O, Grieder TE, Cole M, Koob GF (2010) Exposure to chronic intermittent nicotine vapor induces nicotine dependence. Pharmacol Biochem Behav 96:104–107. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Gilpin NW, Whitaker AM, Baynes B, Abdel AY, Weil MT, George O (2014) Nicotine vapor inhalation escalates nicotine self-administration. Addict Biol 19(4):587–592. CrossRefPubMedGoogle Scholar
  13. Goniewicz ML, Kuma T, Gawron M, Knysak J, Kosmider L (2013) Nicotine levels in electronic cigarettes. Nicotine Tob Res 15:158–166CrossRefGoogle Scholar
  14. Hess CA, Olmedo P, Navas-Acien A, Goessler W, Cohen JE, Rule AM (2017) E-cigarettes as a source of toxic and potentially carcinogenic metals. Environ Res 152:221–225. CrossRefPubMedGoogle Scholar
  15. Javadi-Paydar M, Nguyen JD, Kerr TM, Grant Y, Vandewater SA, Cole M, Taffe MA (2018) Effects of Δ9-THC and cannabidiol vapor inhalation in male and female rats. Psychopharmacology 235(9): 2541-2557. doi: Scholar
  16. Javadi-Paydar M, Kerr TM, Harvey EL et al (2019) Effects of nicotine and THC vapor inhalation administered by an electronic nicotine delivery system (ENDS) in male rats. Drug Alcohol Depend 198:54–62. CrossRefPubMedGoogle Scholar
  17. Jonkman S, Risbrough VB, Geyer MA, Markou A (2008) Spontaneous nicotine withdrawal potentiates the effects of stress in rats. Neuropsychopharmacology 33(9):2131–2138CrossRefGoogle Scholar
  18. Kitzen JM, McConaha JL, Bookser ML, Pergolizzi JV Jr, Taylor R Jr, Raffa RB (2019) e-Cigarettes for smoking cessation: Do they deliver? J Clin Pharm Ther.
  19. Kulik MC, Lisha NE, Glantz SA (2018) E-cigarettes associated with depressed smoking cessation: a cross-sectional study of 28 European Union countries. Am J Prev Med 54(4):603–609. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Leão RM, Cruz FC, Vendruscolo LF, de Guglielmo G, Logrip ML, Planeta CS, Hope BT, Koob GF, George O (2015) Chronic nicotine activates stress/reward-related brain regions and facilitates the transition to compulsive alcohol drinking. J Neurosci 35(15):6241–6253. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Leventhal AM, Strong DR, Kirkpatrick MG et al (2015) Association of Electronic Cigarette Use With Initiation of Combustible Tobacco Product Smoking in Early Adolescence. JAMA 314(7):700–707. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Malin DH, Lake JR, Newlin-Maultsby P, Roberts LK, Lanier JG, Carter VA et al (1992) Rodent model of nicotine abstinence syndrome. Pharmacol Biochem Behav 43:779–784CrossRefGoogle Scholar
  23. Malin DH, Lake JR, Carter VA, Cunningham JS, Hebert KM, Conrad DL, Wilson OB (1994) The nicotinic antagonist mecamylamine precipitates nicotine abstinence syndrome in the rat. Psychopharmacology 115(1-2):180–184CrossRefGoogle Scholar
  24. Matta SG, Balfour DJ, Benowitz NL et al (2007) Guidelines on nicotine dose selection for in vivo research. Psychopharmacology 190(3):269–319CrossRefGoogle Scholar
  25. Miliano C, Scott ER, Murdaugh LB, Gnatowski ER, Faunce CL, Anderson MS, Reyes MM, Gregus AM, Buczynski MW (2019) Modeling drug exposure in rodents using e-cigarettes and other electronic nicotine delivery systems. J Neurosci Methods 12:108458. CrossRefGoogle Scholar
  26. Nguyen JD, Aarde SM, Vandewater et al (2016a) Inhaled delivery of Delta (9)-tetrahydrocannabinol (THC) to rats by e-cigarette vapor technology. Neuropharmacology 109:112–120. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Nguyen JD, Aarde SM, Cole M, Vandewater SA, Grant Y, Taffe MA (2016b) Locomotor stimulant and rewarding effects of inhaling methamphetamine, MDPV, and mephedrone via electronic cigarette-type technology. Neuropsychopharmacology 41:2759–2771. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Nguyen JD, Bremer PT, Hwang CS, Vandewater SA, Collins KC, Creehan KM, Janda KD, Taffe MA (2017) Effective active vaccination against methamphetamine in female rats. Drug Alcohol Depend 175:179–186. CrossRefPubMedPubMedCentralGoogle Scholar
  29. Nguyen JD, Grant Y, Kerr TM, Gutierrez A, Cole M, Taffe MA (2018) Tolerance to hypothermic and antinoceptive effects of ∆9-tetrahydrocannabinol (THC) vapor inhalation in rats. Pharmacol Biochem Behav 172: 33-38. doi: Scholar
  30. O’Dell LE, Koob GF (2007) ‘Nicotine deprivation effect’ in rats with intermittent 23-hour access to intravenous nicotine self-administration. Pharmacol Biochem Behav 86(2):346–353CrossRefGoogle Scholar
  31. Peace MR, Baird TR, Smith N, Wolf CE, Poklis JL, Poklis A (2016) Concentration of nicotine and glycols in 27 electronic cigarette formulations. J Anal Toxicol 40(6):403–407. CrossRefPubMedPubMedCentralGoogle Scholar
  32. Perez-Stable EJ, Benowitz NL, Marin G (1995) Is serum cotinine a better measure of cigarette smoking than self-report? Prev Med 24:171–179CrossRefGoogle Scholar
  33. Singh T, Arrazola RA, Corey CG et al (2016) Tobacco use among middle and high school students–United States, 2011– 2015. MMWR Morb Mortal Wkly Rep 65(14):361–367. CrossRefPubMedGoogle Scholar
  34. Sleiman M, Logue JM, Montesinos VN et al (2016) Emissions from electronic cigarettes: key parameters affecting the release of harmful chemicals. Environ Sci Technol 50(17):9644–9651. CrossRefPubMedGoogle Scholar
  35. Smith TT, Heckman BW, Wahlquist AE, Cummings KM, & Carpenter MJ (2019) The impact of e-liquid propylene glycol and vegetable glycerin ratio on ratings of subjective effects, reinforcement value, and use in current smokers. Nicotine and Tobacco Research.
  36. Torres OV, Gentil LG, Natividad LA, Carcoba LM, O’Dell LE (2013) Behavioral, biochemical, and molecular indices of stress are enhanced in female versus male rats experiencing nicotine withdrawal. Front Psychiatry 4:38. CrossRefPubMedPubMedCentralGoogle Scholar
  37. Turner PV, Brabb T, Pekow C, Vasbinder MA (2011) Administration of substances to laboratory animals: routes of administration and factors to consider. J Am Assoc Lab Anim Sci 50(5):600–613PubMedPubMedCentralGoogle Scholar
  38. Vendruscolo JCM, Tunstall BJ, Carmack SA, Schmeichel BE, Lowery-Gionta EG, Cole M, George O, Vandewater SA, Taffe MA, Koob GF, Vendruscolo LF (2018) Compulsive-like sufentanil vapor self-administration in rats. Neuropsychopharmacology 43(4):801–809. CrossRefPubMedGoogle Scholar
  39. Watkins SS, Epping-Jordan MP, Koob GF, Markou A (1999) Blockade of nicotine self-administration with nicotinic antagonists in rats. Pharmacol Biochem Behav 62(4):743–751CrossRefGoogle Scholar
  40. Weaver SR, Huang J, Pechacek TF, Heath JW, Ashley DL, Eriksen MP (2018) Are electronic nicotine delivery systems helping cigarette smokers quit? Evidence from a prospective cohort study of U.S. adult smokers, 2015–2016. PLoS One 13(7):e0198047. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Werley MS, Kirkpatrick DJ, Oldham MJ, Jerome AM, Langston TB, Lilly PD, Smith DC, Mckinney WJ Jr (2016) Toxicological assessment of a prototype e-cigaret device and three flavor formulations: a 90-day inhalation study in rats. Inhal Toxicol 28(1):22–38. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Physiology, School of MedicineLouisiana State University Health Sciences CenterNew OrleansUSA
  2. 2.La Jolla Alcohol Research Inc.La JollaUSA
  3. 3.Neuroscience Center of ExcellenceLouisiana State University Health Sciences CenterNew OrleansUSA
  4. 4.Alcohol & Drug Abuse Center of ExcellenceLouisiana State University Health Sciences CenterNew OrleansUSA
  5. 5.Southeast Louisiana VA Healthcare System (SLVHCS)New OrleansUSA

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