Acute and subacute (28 days) toxicity of green coffee oil enriched with diterpenes cafestol and kahweol in rats

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Highlights

  • Acute treatment of green coffee oil on female rats did not result in toxic effects nor mortality.

  • The subacute treatment of the green coffee oil promoted weight loss without changes in the food intake at the highest dose.

  • Subacute treatment of green coffee oil decreased cholesterol and glucose levels in all doses studied.

Abstract

Green coffee oil enriched with cafestol and kahweol was obtained by supercritical fluid extraction using carbon dioxide while its safety and possible effects from acute and subacute treatment were evaluated in rats. For acute toxicity study, single dose of green coffee oil (2000 mg/kg) was administered by gavage in female rats. For subacute study (28 days), 32 male rats received different doses of green coffee oil extract (25, 50, and 75 mg/kg/day). In the acute toxicity study, main findings of this treatment indicated no mortality, body weight decrease, no changes in hematological and biochemical parameters, and relative weight increase in heart and thymus, without histopathological alterations in all assessed organs. All these findings suggest that LD50 is higher than aforesaid dose. In the subacute toxicity, main findings showed body weight decrease mainly at the highest dose without food consumption change, serum glucose and tryglicerides levels decrease, and relative weight increase in liver. As evidenced in histopathological pictures, no changes were observed at all treated doses. Our study suggest that green coffee oil can be explored to clinically develop new hypocholesteromic and hypoglycemic agents. However, further studies evaluating long-term effects are needed in order to have sufficient safety evidence for its use in humans.

Introduction

Different oils extracted from green coffee beans (i.e. dried and shell-less coffee) and roasted coffee beans have been studied, with particular interest in the former due to their prospective health effects and benefits (Cornelio-Santiago et al., 2017; De Oliveira et al., 2014, 2018). Using carbon dioxide (CO2) as solvent, coffee oil obtained via supercritical fluid extraction (SFE) has been explored to determine bioactive compounds (Oliveira et al., 2007, 2005, 2009a; 2009b). SFE-extracted green coffee oil has total phenolic compounds (TPC) ranging from 17.02 to 35.99 mg gallic acid equivalent (mg-GAE)/g-extract, which corresponds to 2.36–5.16 mg-GAE/100 g of green coffee beans (on dry basis) (De Oliveira et al., 2018).

This oil also contains cafestol and kahweol, which are diterpenes with high antioxidant activity (Cavin et al., 1998, 2002; De Oliveira et al., 2014, 2018) as well as anti-inflammatory property and cellular protection activity (Frost-Meyer and Logomarcino, 2012; Vignoli et al., 2014). According to Kitzberger et al. (2013), aforesaid diterpenes may prevent liver diseases from developing while exhibiting anticarcinogenic properties (Cavin et al., 1998, 2002; Furtado et al., 2014; Huber et al., 2002).

SFE technology has been applied to green coffee beans for decaffeination purposes as well as to obtain oil rich in those diterpenes (Araújo and Sandi, 2006; Azevedo et al., 2008). In extant and optimized SFE processes, green coffee oil proved to be enriched in cafestol (46.9 g/kg green coffee oil) and kahweol (60.4 g/kg green coffee oil) (De Oliveira et al., 2014, 2018). The information about the toxicity studies of green coffee oil is limited and to the best of our knowledge, the present work is the first study evaluating possible toxic effects from both acute and sub-acute toxicity in rats treated with green coffee oil enriched in diterpenes cafestol and kahweol via SFE.

Section snippets

Animals

Wistar male and female 7-8-week-old rats were maintained in accordance with the Guide for the Care and Use of Laboratory Animals, National Research Council, USA (NRC, 2010). All experiments were performed in accordance with good laboratory practice protocols as well as quality assurance methods. Six female nulliparous and non-pregnant rats and 34 male rats, respectively weighting 130–150 g and 180–200 g, were respectively used for acute and subacute experiments (Ameni et al., 2015). All animals

Acute oral toxicity

Administration of a single oral dose of 2000 mg/kg of green coffee oil extract did not result in any sign of toxicity in animals during the observation period (14 days). Regarding body weight, repeated measure showed differences on days of observation (F(14,28) = 7.65, p < 0.001), without differences between treatment (F(1,2) = 0.08, p = 0.80) or neither interaction (F(14,28) = 2.026, p < 0.054). Dunnett's multiple comparisons test showed body weight decrease on days 1 (p < 0.05), 2 (p < 0.01),

Discussion

In the present study, green coffee oil rich in diterpenes extracted via SFE was used for evaluated acute and sub-acute oral toxicity in rats. In the acute toxicity study (2000 mg/kg/bw), the main findings of this treatment indicated no mortality; no significant effects in feed consumption; decrease in the body weight (after 4 days of treatment); no changes in hematological and biochemical parameters; increased in relative heart and thymus weights without histopathological alterations. In the

Conclusion

This study demonstrated that oral administration of green coffee oil at a single dose of 2000 mg/kg showed no mortality or signs of toxicity, indicating that LD50 is higher than this dose. Additionally, in the acute toxicity study no changes in hematological and biochemical parameters were observed while an increased in relative weight of heart and thymus were observed, without histopathological alterations in these organs. In subacute toxicity study (28 days), body weight decrease was observed

Funding body information

Authors acknowledge the funding from Sao Paulo State Research Foundation (FAPESP), process number 2013 / 03371-0.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

Authors would like to thank FAPESP (State of São Paulo Research Foundation, Brazil) for financial support under the research project for the MSc scholarship (FAPESP nº 2013/03371-0).

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  • Cited by (0)

    1

    These authors contributed equally to this work.

    2

    Present address: Department of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, Canada.

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