L-(−)-Menthol (Fisher Scientific, Atlanta, Georgia, USA) was dissolved in 50% dimethyl sulfoxide (DMSO; Sigma; Atlanta, Georgia, USA) and distilled water (v/v). Menthol and vehicle (50% DMSO: H2O, v/v) solutions were freshly prepared weekly and injected intraperitoneally (IP) at 1 ml/kg with a 5-min IPI. All menthol and vehicle solutions were heated to 50°C before each session to ensure menthol solubility. All nicotine and menthol training doses were selected based on previous literature (Biswas et al., 2016 (link); Huynh et al., 2019 (link)). (−)-Nicotine bitartrate dihydrate (MP Biomedicals; Solon, Ohio, USA) was dissolved in 0.9% saline, titrated with NaOH to a pH of 7.0 ± 0.2, and injected subcutaneously (SC) at 1 ml/kg with a 5-min IPI. All nicotine doses are expressed as the free base form and all menthol doses are expressed as the salt form.
Protocol detail
Menthol and Nicotine Pharmacological Effects
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Other organizations : University of Nebraska–Lincoln
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Variable analysis
independent variables
- Menthol concentration (50% DMSO: H2O, v/v)
- Nicotine concentration (0.9% saline, pH 7.0 ± 0.2)
- Not explicitly mentioned
- Injection volume (1 ml/kg)
- Injection interval (5-min IPI)
- Menthol and vehicle solution preparation (freshly prepared weekly, heated to 50°C)
- Positive control: Not explicitly mentioned
- Negative control: Vehicle (50% DMSO: H2O, v/v)
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