Research Article

Antimicrobial activities of volatile oil compound from the rhizome of Zingiber officinale using solvent method extraction

Ugoh Sylvanus1, Fatokun Olakunle 2, Olajide Olutayo2*, Kabiru Usman1

1Department of Biological Sciences, University of Abuja, Nigeria

2Chemistry Advanced Laboratory, Sheda Science and Technology Complex, Sheda Abuja, Nigeria

*For correspondence

Olutayo Olajide

Chemistry Advanced Laboratory, Sheda Science and Technology Complex, Sheda Abuja, Nigeria

Tel:+23407033477293

Email:olajidewumi@rocketmail.com

Received: 15 Apr 2015

Accepted: 24 Jun 2015

ABSTRACT

Objective: To characterize the chemical constituents and evaluate the antimicrobial activity of Zingiber officinale oil extracted with hexane.

Methods: The air dried powdered rhizome of Z. officinale was extracted with n- hexane. The antimicrobial activity of Z. officinale oil extract was tested on gram positive and gram negative bacteria.

Result: The extracted oil on GC-MS analysis resulted in the identification of 19 compounds, among which 13-Docosenoic acid was found to be the most abundant at 15.48 %. The preliminary antimicrobial screening of the oil from Z. officinale against four pathogens; Pseudomonas aeruginosa, Escherichia coli, Salmonella typhi and Staphylococcus aureus revealed that the mean zone diameter of inhibition (Z.O.I) is concentration dependent in which pseudomonas aeruginosa showed highest concentration at 100 %. The minimum inhibitory concentrations (MIC) values of the oil revealed these concentrations 12.5 %, 25 % 50 % and 100 % against the tested organisms respectively. The minimum bactericidal concentration (MBC) of the oil on S. aureus was recorded at 100% concentration, while E. coli, P. aeruginosa, and S. typhi were all resistant.

Conclusion: This study showed that the oil from Z. officinale may be used to manage some infectious diseases caused by the tested organisms.

Keywords: Zingiber officinale, Chemical constituents, Antimicrobial activities, Medicinal uses

Introduction

Traditionally uses of medicinal plants have recently attracted the attention of pharmaceuticals and scientific communities which had involved the isolation and identification of secondary metabolites produced by plants and their uses as active principles in medicinal properties[1].

Zingiber officinale commonly referred to as ginger is a perennial herb and grows to about 3 - 4 feet high with a thick spreading tuberous rhizome, the leaves are spear-shaped with white or yellow flowers growing directly from the root[2]. Since ancient time Z. officianale had been used in treating arthritis, colic, diarrhea and heart related problems[3]. The pharmacological activities can be attributed to the presence of the followings: α-pinene, camphene, β-pinene, 1, 8-cineole, linalool, borneol, γ-terpineol, nerol, neral, geraniol, geranial, geranyl acetate, β-bisabolene and zingiberene[4]. The main principles in Z. officianale are the series of pungent oleoresin constituents known as gingerol being the major component[1] 6-gingerol and 6-shagaol[3]. Many reports are available on the chemical composition of ginger oil and the natural occurring flavouring compounds[4]. Ginger contains 1-2 % oil that impacts the unique flavour to the spice[5] Z. officinale is widely used in ayurvedic medicines and folklore medicines, several spices and herbs exert antibacterial influences due to the essential oil fraction. It acts as antioxidants that protect against cancer, arteriosclerosis, heart disease and several other diseases[3]. Z. officinale owes its aroma to about 1 to 3 % of volatile oils; these are bisabolene zingiberene and zingiberol[6]. The oil can vary in color from pale yellow to darker amber colour and the viscosity also ranges from medium to watery. It is sharp, warm and strong smelling spicy oil. The study traces the antimicrobial activity and the chemical constituents of the solvent extract of Z. officianale oil.

Materials and Methods

Collection of Sample

The ginger was bought from spices sellers in Gwagwalada market, Abuja. The Voucher /herbarium number has been deposited at the Department of Biological Sciences, University of Abuja.

Extraction of volatile oil from Z. officinale

The plant material was pulverized into powder using the laboratory mortar and pestle. It was kept in a sterile polythene bag and labeled. Solvent extraction 2 kg of cut ginger were mixed with 2 litres of solvent (n-hexane). The mixture was kept for 24 hours. The homogenate was filtered using what man No. 4 filter paper. The solvent was evaporated with a rotary evaporator at 40oC[7].

Gas chromatography- mass spectrometry analysis

The hexane extract from Z. officianale was analyzed on a GC-MS SHIMADZU. GCMS-QP2010 PLUS using an OV- 1701 column (25 m × 0.25 mm, ø with 0.15 μm film thickness) was used Helium was the carrier gas at 1.2 ml/min. The MS operated on an electron mode (70 eV), ion source 2300C. The percentage compositions of the constituents of the oil were computed in each case from GC peak areas. The identification of the components was based on the comparison of retention indices (determined relative to the retention time of series of n-alkanes) and mass spectral with those of authentic samples (NIST 05.L database/chemstation data system) and with data from literature[8,9].

Biological screening of extractives

The oil extract was screened for antimicrobial activity using out using agar well diffusion technique with little modification[3]. Both sensitivity and minimum inhibitory concentrations (MIC) were determined.

Determination of Minimum Bactericidal Concentration (MBC)

Minimum bactericidal concentration (MBC) was determined by taking 0.1 ml of the content of the tubes of the MIC showing no growth and inoculating it into sterile nutrient agar. These plates were then incubated at 37°C for 24 h.

Results and Discussion

The GC-MS analysis revealed the presence of nineteen compounds from the oil of Z. officinale extracted with n- hexane (Table 1). The major constituents of the oil include α –Zingerone (8.36 %), 13-Docosenoic acid (15.48 %), n-Decane (4.468 %). The most abundant compound, 13-Docosenoic acid is in agreement with earlier report[10].

Table 1: Chemical constituents of the volatile oils of Z. officinale based on GC-MS analysis.

S. No.

Names

RT

% Yield

(M+) BP

1

n-Nonane

3.385

4.37

(128) 43

2

2,3-Dimethyl-2-hexanone

3.731

2.55

(128) 43

3

n-Decane

4.468

7.63

(142) 43

4

1,2,3-Trimethylbenzene

4.680

8.50

(120)105

5

n-Undecane

5.753

2.54

(156) 43

6

1,2,4-Trimethyl-3-nitrobicyclo[3.3.1]nonan-9-one

7.224

3.29

(225) 95

7

Cis-Farnesol

8.267

1.48

(222) 41

8

1-(1-Hydroxy-2,6,6-trimethyl-2,4-cyclohexadien-1-yl)ethanone

10.579

6.82

(180) 43

9

α –Zingerone

11.388

8.36

(204)119

10

β-Farnesene

11.520

2.68

(204) 41

11

β-Sesquiphellandrene

11.757

4.28

(204) 41

12

Δ-nerolidol

12.213

1.87

(222) 41

13

Zingerone

13.879

2.23

(194) 43

14

Palmitic acid

17.663

3.69

(256) 43

15

Isomyreceneyl acid

18.506

1.76

(196) 43

16

13-Docosenoic acid

21.281

15.48

(210) 55

17

9-Hexadecenol

21.574

5.97

(240)41

18

Oleic acid

25.325

3.70

(282) 55

19

Linoleic acid chloride

25.325

9.45

(298) 55

071215_1913_Antimicrobi1

Figure 1: Chemical constituents of the volatile oils of Zingiber officinale based on GC-MS analysis.

Key: R.T. =Retention Time; (M+) = Molecular ion; BP = Base Peak

Essential oils are complex mixtures comprising many single compounds, each of the constituents contribute to the biological effects of these oils[11]. α–Zingeberene is a monocyclic sesquiterpene, a major constituent of ginger oil that impacts the distinct flavoring.

13-Docosenoic acids are found in rapeseed, wallflower seed, and mustard seed, making up 40-50% of their oils[12]. It possesses same application as mineral oils. Its industrial use are quite numerous; it is suitable for transmission oil, a binder for oil paint, it readily forms many organic compounds, suitable for use as organic matrices that need to be polymeric, valued in tribology as a superior lubricant, it is variously used as emollients, especially for skin and healthcare products. Being a hydrocarbon of high calorific value, with a very low flash point, high cetane number and good lubrication qualities it can serve as valuable component of biodiesel[13]. It is widely used to produce emollients, especially for skin and healthcare products. The antimicrobial screening (Table 2) of the Z. officinale exhibited activity against the tested organisms: S. aureus, S. typhi, P. aeruginosa and E. coli; S. typhi. P. aeruginosa had the highest value of 35.00 mm at 100 % and it decreases significantly as the concentrations decreases to 8.00 mm 12.5%, which is in agreement with previous study[14]. Also previous studies[15] indicated that E. coli, S. typhi and P. aeruginosa are inhibited by the essential oil of Z. officinale which is in agreement with the oil extracted from the same plant using n-hexane.The entire organisms were susceptible as evidenced by the mean zone of inhibition compared with the standard drug (Ciprofloxacin) which is suggestive that the chemical constituents of Z. officinale possess antibacterial properties[6]. Further antimicrobial screening of the oil revealed the following The MIC values at 12.5 %, 25 %, 50 % and 100 % against S. aureus, P. aeruginosa, S. typhi and E. coli respectively. The MBC showed the activity of S. aureus at 100 % only.

Table 2: The mean diameter zone of Inhibition of Z. officinale essential oil.

Test organisms   Concentration  
  12.5: 87.5 25 : 75 50 : 50 100 : 0 Control
E. coli 2.50± 0.50 11.00± 1.00 17.00± 1.00 23.50 ±2.50 33.50 ±0.50
S. aureus 10.50± 2.50 20.50± 0.50 26.00± 1.00 34.00 ±2.00 31.00 ±1.00
P. aeruginosa 17.00± 1.00 22.00± 1.00 25.50± 0.50 35.50 ±2.50 28.00 ±1.00
S. typhi 8.50± 2.50 15.50± 3.50 27.00± 1.00 33.00 ±2.00 30.50 ±1.50

Table 3: The minimum inhibitory concentration of oil of Z. officinale oil.

test organisms Concentrations 100
12.5 25 50
E. coli + + + -
S. aureus + - - -
P. aeruginosa + + - -
S. typhi + - - -

Table 4: The minimum bactericidal concentration (MBC) of essential oil of ginger.

test organisms Concentration 100
12.5 25 50
E. coli + + + +
S. aureus + + + -
P. aeruginosa + + + +
S. typhi + + + +

Keys: + = Growth, - = No growth

Conclusions

The trend observed in this study supports the traditional use of Zingiber officinale oil in the treatment of infections and therefore can be used for the development of broad spectrum antibiotics. However further studies should be carried out to assess the active components of the oil which have specific antimicrobial activity.

Funding: No funding sources

Conflict of interest: None declared

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