Journal of
Medicinal Plants Research

  • Abbreviation: J. Med. Plants Res.
  • Language: English
  • ISSN: 1996-0875
  • DOI: 10.5897/JMPR
  • Start Year: 2007
  • Published Articles: 3843

Full Length Research Paper

Chemical Composition antimicrobial and free radical scavenging activity of essential oil from leaves of Renealmia thyrsoidea (Ruiz &Pav.) Poepp. & Endl.

Paco F. Noriega
  • Paco F. Noriega
  • Centro de Investigación y Valoración de la Biodiversidad, Universidad Politécnica Salesiana, Quito-Ecuador.
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Erika A. Paredes
  • Erika A. Paredes
  • Centro de Investigación y Valoración de la Biodiversidad, Universidad Politécnica Salesiana, Quito-Ecuador.
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Tatiana D. Mosquera
  • Tatiana D. Mosquera
  • Centro de Investigación y Valoración de la Biodiversidad, Universidad Politécnica Salesiana, Quito-Ecuador.
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Edison E. Diaz
  • Edison E. Diaz
  • Symrise AG. Mühlenfeldstraße 1 37603 Holzminden, Germany.
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Angelika Lueckhoff
  • Angelika Lueckhoff
  • Symrise AG. Mühlenfeldstraße 1 37603 Holzminden, Germany.
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Jessica E. Basantes
  • Jessica E. Basantes
  • Centro de Investigación y Valoración de la Biodiversidad, Universidad Politécnica Salesiana, Quito-Ecuador.
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Andrea L. Trujillo
  • Andrea L. Trujillo
  • Centro de Investigación y Valoración de la Biodiversidad, Universidad Politécnica Salesiana, Quito-Ecuador.
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  •  Received: 02 August 2016
  •  Accepted: 25 August 2016
  •  Published: 03 September 2016

 ABSTRACT

Renealmia thryrsoidea  (Ruiz & Pav.)  Poepp. & Endl is a plant used by the Amazonian indigenous people of Ecuador for its various medicinal properties. Its leaves exhibit a remarkable aroma with a hint of spiciness. The essential oil extracted from its leaves was analyzed by GM/MS, using two systems with columns of different polarity, in both was confirmed  the presence of terpinolene (26.32%), α-felandrene (17.16%), γ-terpinene (6.55%), β-pinene (5.97%) and p-cymol (4.70%). Free radical scavenging activity was analyzed through 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) spectrophotometric methods ; by applying ABTS method , the activity was comparable to that of the essential oil of Thymus vulgaris. The trials of antimicrobial activity show a strong inhibition against  Gram negative bacteria as  Escherichia coli and Pseudomonas aeruginosa. 
 
Key words: Renealmia  thryrsoidea, essential oil, DPPH, ABTS, minimum inhibitory concentration (MIC).
 


 INTRODUCTION

Essential oils are a group of secondary metabolites that currently have a productive significance (Schmidt, 2015). Many of them are valued for their qualities as potential medicinal products (Noriega, 2009) and cosmetics (Muñoz-Acevedo et al., 2015). Ecuador is a country with a very high plant diversity (Bendix et al., 2013) and many of these species are rich in essential oils that have various medicinal uses (Malagon et al., 2003; Sacchetti et al., 2005; Guerrini et al., 2009). 
 
The Renealmia genus (Zingiberaceae family) is distributed throughout South America with 75 species (Maas, 1997). In Ecuador, the number of species reaches 25, 4 of which are endemic (Jorgensen and León-Yanez, 1999).
 
Renealmia thyrsoidea (Ruiz & Pav.) is a widely distributed species in tropical America, in countries such as Bolivia, Colombia, Ecuador, Guyana, Nicaragua, Panama, Peru, Suriname, Trinidad and Tobago and Venezuela. In Ecuador this spices is distributed in the three continental regions: the coast, the highlands, and Amazon; with a noted presence in the provinces of Cotopaxi, Imbabura, Morona Santiago, Napo, Pastaza, 
Pichincha, Sucumbios, Tungurahua and Zamora Chinchipe (Jorgensen and León-Yanez, 1999).  An important reported use is that of acting as an antidote for snake bites (Davis and Yost, 1983). The antimalarial and antipyretic properties of R. thyrsoidea were reported by Schultes et al. (1990) and Céline et al. (2009), this plant possesses analgesic and anti-flu properties (de La Torre et al., 2008), and anti-leishmaniasis uses are reported in various regions (Yannick, 2009). The fruits are edible (de la Torre et al., 2008) and a dye is extracted from them to be used in ritual practices (Maas, 1997). 
 
Several of the ethnic groups present in the Amazon region of Ecuador employ the species. The importance of this research lies in the assessment of the plant’s essential oils for pharmacological and medicinal traits.

 


 MATERIALS AND METHODS

Plant
 
The leaves of R. thyrsoidea were collected in the community of San Luis de Inimkis, Canton Macas, province of Morona Santiago, with the following geographic coordinates: 02° 26' South latitude and 78° 11' West longitude; at an altitude of 1070 m.o.s.l. The botanical identification took place in the herbarium Alfredo Paredes of the Universidad Central del Ecuador by the botanist Carlos Cerón. The fresh leaves were collected from mature plants and distilled in vapor stream, with an equipment of 250 liters capacity belonging to the Fundacion Chankuap recursos para el futuro, in the city of Macas. 
 
Chemical composition
 
The essential oil composition was determined using gas chromatography and mass spectrometry equipped with two columns of different polarity systems. The sample was prepared by dissolving 25 µl of essential oil in 1 ml of acetone. For coupling 1, a gas chromatograph GC Agilent 7890 A was coupled to a mass spectrometer MS 5975. The column used was a DB1-MS, with a length of 60 m, 0.32 mm internal diameter and 0.25 um thickness. The analysis started at a temperature of 60°C with a rate of 4°C per min up to 280°C. The carrier gas was helium at a flow of 3.5 ml min-1. The injection volume was 1 μl. For coupling 2, a gas chromatograph GC Agilent 6890 A was coupled to a mass spectrometer MS 5973. The column used was a DB-Wax, with a length of 30 m, 0.25 mm internal diameter and 0.25 um thick. The analysis started at a temperature of 60°C with a velocity gradient of 3°C per min up to 240°C. The carrier gas was helium at a flow of 3.3 ml min-1. The injection volume was 1 μl. In both cases, electron ionization was used with energy of 70 eV and a range of m/z 35 to 400 Da. The component identification was performed by comparing the mass spectra to those present in the MASSLIB database and by determining the retention index for each component using as reference the standard series C8-C30 of hydrocarbons.
 
Free radical scavenging activity
 
Spectrophotometric methods DPPH (diphenyl-2-picrylhydrazyl) and ABTS [2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt] have been a permanent technique to evaluate the free radical scavenging activity in essential oils (Bardaweel et al., 2015; Ornano et al., 2015). Fort the DPPH assay, varying amounts of R. thyrsoidea essential oil were taken and dissolved  in dimethyl sulfoxide (DMSO) to a volume of 100 µl. To each solution, 2.9 ml of DPPH (1 × 10-4 molar in ethanol) was added. The solution was stirred vigorously for 30 min in the dark at room temperature. The absorbance was measured at 517 nm in a Shimadzu UV 1240 mini. Similarly, the analysis proceeded with the ABTS test, to each solution dissolved in DMSO 0.9 ml of ABTS 40 mM (previously radicalized with a K2S2O8 solution) was added. The absorbance was measured at 734 nm in a Shimadzu UV mini 1240. Antiradical activity for each mixture was calculated according to the following formula:
 
 
Where Aa and Ab are the absorbance of blanks and samples, respectively after 30 min (DPPH) and 1 min ABTS. The activity of the essential oil was evaluated by calculating the IC50, which is 50% inhibition of oxidation of DPPH and ABTS; this was calculated from the data of the calibration curves obtained from the data of the concentration versus percentage inhibition. As referents of activity, Tymus vulgaris essential oil and butylated hidroxianisole (BHA) were used. 
 
Antimicrobial activity (MIC evaluation)
 
The antimicrobial activity was evaluated using the disc diffusion method often described as an applicable methodology to evaluate the antimicrobial activity of essential oils (Rivera et al., 2015; Pesavento et al., 2015). The strains tested were Grampositive bacteria: Staphylococcus aureus subsp. aureus ATCC 6538 and Streptococcus mutans ATCC 25175; Gramnegative bacteria: Escherichia coli ATCC 8739 and Pseudomonas aeruginosa ATCC 9027. The yeasts Candida tropicalis ATCC 13803 and Candida albicans ATCC 10231 were also tested. The antimicrobial activity is described as the minimum inhibitory concentration (MIC) in mg ml-1. The essential oil of T. vulgaris was used as a reference for any appreciable activity (Rossi et al., 2011).
 
 
 
 
 


 RESULTS AND DISCUSSION

Production of the essential oil 
 
The essential oil yield was 0.047% (w/w). Essential oil density was 0.873 mg ml-1.
 
Chemical composition
 
In both studies with both DB1 and DB-Wax column the most abundant compounds were: terpinolene (26.32%), α-felandrene (17.16%), γ-terpinene (6.55%), β-pinene (5.97%) and p- cymol (4.70%). The analysis with DB1 column reveals the presence of components 116; DB-Wax column revealed the presence of 107 compounds with the same equivalent percentages, as shown in Table 1.
 
Free radical scavenging activity
 
IC50 values indicate the ability to inhibit by 50%, the concentration of free radicals present. For this test, the reference essential oil was that of T. vulgaris whose thymol and carvacrol components are widely known for this property (Mastelić et al., 2008; Lee et al., 2005). The results are shown in Table 2.
 
 

Antimicrobial activity

 

There are different results depending on the microbial strains evaluated. A good activity was observed in the Gram negative bacteria tested, close to the natural pattern registered with the essential oil of T. vulgaris, represented by the values in Table 3.

 

 
 
 
 
 
 
 


 CONCLUSION

The study provides an insight into the chemical components present in the essential oil from the  leaves  of R. thyrsoidea. Most notable was the presence of monoterpenes. As for the antioxidant activity appreciable activity highlighted in the evaluation of electron scavenging capacity ABTS methodology, whereas by the method DPPH oil is less active. Interestingly, anti-microbial activity evaluated in both Gram negative strains, whose values ​​of minimum inhibitory concentration reaches 0.35 mg ml-1.
 

 


 CONFLICT OF INTERESTS

The authors have none to declare.


 ACKNOWLEDGEMENTS

We would like to thank Symrise for funding and supporting this research.



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