Chemical Composition and Antibacterial Activity of Essential Oil of Mentha Longifolia Leaf from Albaha Area Southern Saudi Arabia

Burham BO, Mohammed Nour AA and Osman OA

Published on: 2019-04-26

Abstract

The essential oil obtained by hydrodistillation from the leaves of Saudi Arabia native Mentha longifolia (Lamiaceae) was analyzed by gas chromatography-mass spectrometry GC/MS. Forty six compounds were analysed , it was characterized by a high percentage of oxygenated monoterpenes (30.40 %) with Piperitone as the major one (30.77 %), followed by sesquiterpene hydrocarbons (26.08%) in which Caryophyllene (5.58%) as the main, while monoterpene hydrocarbons (23.90%) with gamma-Terpinene (1.36%) as a main. The principal components of M. longifolia oil was the piperitone with chemical formula C10H14O. The antibacterial activity of the essential oil of M. longifolia L. leaf were evaluated in this study. The oil was screened against Gram positive (Staphylococcus aureus, Bacillus subtilis) and Gram negative bacteria (Escherichia coli, Pseudomonas aeruginosa). The essential oil showed strong antibacterial activity against Bacillus subtilis with inhibition zone (25 mm), and Escherichia coli (24 mm), It seems that the essential oils derived from the Mentha longifolia L. oil could be used as a natural source of antimicrobial agents.

Keywords

Mentha longifolia GC/MS analysis Essential oil Antibacterial Saudi

Introduction

Medicinal and aromatic plants constitute a large part of the natural flora and are considered an important resource in various sectors such as pharmaceuticals, flavor and fragrance, perfumery and cosmetics [1]. In medicinal and aromatic plants, essential oils generally accumulate in secretory canals or cavities and in glandular trichomes and sometimes in epidermal cells [2]. Essential oils and their chemical constituents show greater bioactivity when they are present in the oxygenated or active form. In general, the chemical composition of essential oils is relatively complex and 20 to 60 different bioactive components are observed in many of these essential oils. Natural products are the source of synthetic and traditional herbal medicine. About 80% of the world population uses traditional medicine, which has compounds derived from medicinal plants [3] these plants are also used to stabilize different food products from deterioration. They have contributed significantly to the treatment of major body disorders. Traditional folk plant remedies have always driven scientists to search for new drugs to maintain and promote healthy living for humans and animals [4]. The therapeutic efficacy of many indigenous plants for various diseases has been described by traditional herbal doctors [5]. Wild mint is a traditional folk medicine. It is mainly used for respiratory disorders, coughs, colds, stomach cramps, asthma, flatulence and headaches [6-7]. Mentha ssp. has been used as a folk remedy for the treatment of nausea, bronchitis, flatulence, anorexia, ulcerations, colitis and liver disorders due to its anti-inflammatory, carminative and antioxidant activities [8]. Externally, they were applied to treat acne, odor loss, insect bites, snake bites and skin infections [9].The activity is mainly attributed to a variety of phenolic compounds and to the composition of essential oil. The main compounds responsible for the typical aroma are chavicol methyl ether (estragol), linalol, eugenol, 1, 8-cineol and methyl cinnamate [10]. The family is also famous for the presence of diterpenoids in its members.

However, the biological activity of the oil depends on the chemical constituents present in the oil, which vary greatly depending on the geographical region. For example, essential oil rich in piperitone oxide (from Lithuania and Jordan), carvone and cis-carveolo (from Iran), pulegone (from Israel) and diosphenol (from Spain) [11-16] have been reported. The present study aimed to examine the chemical composition of M. Longifolia such as essential oil of leaves to show its importance for industrial (i.e., medical and pharmaceutical) purposes.

Materials and Methods

Plant material

Mentha longifolia was obtained from a local market in AL-Baha city and authenticated by D. Haidar Abd-Elgadirin AL-Baha University, Faculty of Science, department of Biology

Isolation of the essential oil

The dried sample of leaves was submitted to hydrodistillation for 8 hours using Clevenger type apparatus [17] according to European Pharmacopoeia. The collected oil was dried over anhydrous sodium sulphate and stored in sealed vials at 15 C until analysis.

Gas Chromatography - Mass Spectrometry (GC/MS)

The qualitative and quantitative analysis of the oil sample was carried out by using GC/MS technique model (GC/MS-QP2010-Ultra) from Japan Shimadzu Company, with serial number 020525101565SA and capillary column (Rtx-5ms-30m x 0.25mm x 0.25um). The sample was injected by using split mode, instrument operating in EI mode at 70eV. Helium as the carrier gas passed with flow rate 1.69 ml/min, the temperature program was started from 50 C with rate 7C/min to 180 C then the rate was changed to 10 C/min reaching 300 C as final temperature degree, the injection port temperature was 300 C, the ion source temperature was 200 C and the interface temperature was 250C. The sample was analyzed by using scan mode in the range of m/z 40-500 charges to ratio and the total run time was 30 minutes.

Identification of components

Identification of components for the sample was achieved by comparing their retention times and mass fragmentation patterns with those available in the library, the National Institute of Standards and Technology (NIST), results were recorded.

Antimicrobial activity

Antibacterial activity was assessed by using a wide range of Gram-positive and Gram-negative bacteria. The zone of inhibition against the microbes was determined using disc diffusion method. Microorganisms were grown in nutrient broth medium and incubated, with shaking, at 37 °C and at 33 °C, for bacteria.

Results And Discussion

The components in essential oil in the leaves of M. longifolia are presented in Table 1. The essential oil obtained from leaves of M. longifolia growing in Saudi. GC/MS analysis of essential oil revealed the presence of 46 different compounds (Table 2). The retention time RT, molecular mass and the relative percentages of the compounds present in leaves of M. longifolia were recorded. The GC/MS chromatogram spectrum confirmed the present compounds. Interpretation of mass spectrum GC/MS was conducted using the database of National Institute Standard and Technique NIST). The oil is characterized by a high concentration of monoterpenes (81.46%) including mostly oxygenated monoterpenes (75.63%). Concerning the high content of sesquiterpene hydrocarbons, 14.14%. The prevailing compounds are piperitone (30.77%), eucalyptol (14.85%), C10H16O (13.68%) and caryophyllene (5.58%). The essential oil chemistry of Mentha longifolia has been studied by many researchers [18-22]. They reported that the major components were cis-piperitone epoxide, pulegone, piperitone oxide, menthol, menthone, menthyl acetate, eucalyptol and piperitone. Most essential oils are composed of terpenes, terpenoids, and other aromatic and aliphatic constituents with low molecular weights. Terpenes or terpenoids are synthesized within the cytoplasm of the cell through the mevalonic acid pathway [23]. Terpenes are composed of isoprene units and are generally represented by the chemical formula (C5H8)n. Terpenes can be acyclic, monocyclic, bicyclic, or tricyclic [24]. Terpenes are classified into several groups such as monoterpenes (C10H16), sesquiterpenes (C15H24), diterpenes (C20H32), and triterpenes (C30H40).

The main component (over 90%) of bioactive essential oils is made up of monoterpenes [25]. Some of the main compounds include monoterpenes hydrocarbons (limonene, -Pinene, p-cymene and -terpinene), oxygenated monoterpenes (thymol, carvacrol, eugenol and camphor), diterpenes (kaurene and camphor), sesquiterpene hydrocarbons (caryophyllene, germacrene D and humulene), oxygenated sesquiterpenes (spathulenol, caryophyllene oxide), monoterpenes alcohols (geraniol, linalol and nerol), sesquiterpene alcohol (patchoulol), aldehydes (citral, cuminal), acids (geranic acid, benzoic acid), ketones (acetophenone, benzophenone) , lactones (bergaptene), phenols (eugenol, thymol, carvacrol and catechol), esters (bornyl acetate, ethyl acetate) and coumarins (fumarin, benzofuran) [1,2,26-29]. The essential oil of leaves from M. longifolia was examined for its antibacterial activity potential against a panel of pathogenic microorganisms including Gram positive, Gram negative, (Table 3). M. longifolia demonstrated variable level of antimicrobial activity against all examined microorganisms.

Essential oil extracted from Mentha longifolia oil showed the highest activity against Bacillus subtilis and Escherichia coli with the strongest inhibition zone of 25 and 24 mm respectively, these in agree with the study by Hafedn et al [30], they proved that the essential oil of M. longif oil a has shown interesting anti-microbial activity against E.coli, S. typhimurium.

The most effective antibacterial activity was expressed by the essential oil against the Gram-negative bacteria, escherichia coli. M D I Z = Mean diameter of growth inhibition zone = average of two replicates in millimeters Inhibition zone: 10-12 weak, 13-15 moderate, 16-18 strong, ≥ 18 highly strong.

Standard Bacteria

Staphylococcus aureus (ATCC 25923), Bacillus subtilis (NCTC 8236). Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853).

Conclusion

By isolating and identifying the compounds, new drugs can be formulated, after pharmacological studies, to treat various diseases.

Recommendations

A number of compounds, representing a variety of chemical compounds isolated from the various plant species, may provide interesting leads for further pharmacological considerations.

Acknowledgement

The authors are highly grateful to Professor Khaled A. Elshafeek (Department of chemistry, Albaha University) for his support during conducting the study.

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Tables

Table 1: Composition of the essential oil obtained from the leaf of M. longifolia.

Peak #

Compound

RT (min)

Area (%)

Molecular mass

Chemical formula

1

alpha-thujene

4.565

0.03

136

C10H16

2

Alpha-pinene

4.697

0.56

136

C10H16

3

Camphene

4.970

0.12

136

C10H16

4

(+)-Sabinene

5.399

0.57

136

C10H16

5

2(10)-pinene

5.475

1.21

136

C10H16

6

Beta-Myrcene

5.682

0.26

136

C10H16

7

(+)-2-Carene

6.203

0.83

136

C10H16

8

O-Cymene

6.366

0.09

134

C10H14

9

D-Limonene

6.442

0.48

136

C10H16

10

Eucalyptol

6.512

14.85

154

C10H18O

11

gamma-Terpinene

7.033

1.36

136

C10H16

12

Cis-sabinene hydrate

7.244

0.71

154

C10H18O

13

Cyclohexene,4-methyl

7.634

0.32

136

C10H16

14

Linalool

7.860

1.30

154

C10H18O

15

Cis-para-menth-2-en-1-ol

8.349

0.22

154

C10H18O

16

Trans-(-)-pinocarveol

8.730

0.54

152

C10H16O

17

Cis-Verbenol

8.843

0.20

152

C10H16O

18

Pinocarvone

9.199

0.33

150

C10H14O

19

Isoborneol

9.289

3.45

154

C10H18O

20

(-)-Terpinen-4-ol

9.491

3.48

154

C10H18O

21

alpha-terpineol

9.769

3.88

154

C10H18O

22

Unidentified

11.051

13.68

152

C10H16O

23

D-carvone

11.401

1.93

150

C10H14O

24

2-pinen-7-one

11.820

0.29

150

C10H14O

25

Beta-elemene

12.600

0.77

204

C15H24

26

Piperitone

12.788

30.77

150

C10H14O

27

Isopropylidenecyclohexanone

13.222

0.79

138

C9H14O

28

(-)-beta-Bourbonene

13.547

0.18

204

C15H24

29

Unidentified

13.645

0.28

204

C15H24

30

Caryophellene

14.205

5.58

204

C15H24

31

alpha-Cubebene

14.663

0.14

204

C15H24

32

Cis-beta-Farnesene

14.735

0.14

204

C15H24

33

Humulene

14.823

0.31

204

C15H24

34

1H-Cyclopenta[1,3]cyclopropa[1,2] benzene

14.983

0.21

204

C15H24

35

1,6-Cyclodecadiene,1-methyl cyclodecadiene

15.312

3.84

204

C15H24

36

Germacrene B

15.589

1.74

204

C15H24

37

gamma-Muurolene

15.880

0.57

204

C15H24

38

Gamma-cadinene

16.015

0.20

204

C15H24

39

Spatulenol

17.031

0.32

220

C15H24 O

40

Caryophellene oxide

17.127

0.28

220

C15H24O

41

Cubenol

17.630

0.17

222

C15H26O

42

tau-Cadinol

18.047

1.88

222

C15H26O

43

alpha-Cadinol

18.283

0.11

222

C15H26O

44

Prasterone-3-sulfate

23.587

0.44

368

C19H28 O 5

45

Unidentified

23.954

0.45

308

C20H36O2

46

Denatonium saccharide

27.660

0.15

446

C28H34N2O3

 Table 2: Components of essential oil from the leaf of M. longifolia.

Compounds

%

Oxygenated Monoterpenes (OM)

30.4

Sesquiterpene Hydrocarbons (SH)

26.08

Monoterpene Hydrocarbons (MH)

23.9

Oxygenated Sesquiterpenes (OS)

10.89

Table 3: Antibacterial Activity of M. longifolia oil (leaves).

Bacteria

Concentration  (mg/ml)

100

50

25

12.5

Staphylococcus aureus

14

13

15

14

Bacillus subtilis

25

21

22

18

Escherichia coli

24

22

21

20

Pseudomonas aeruginosa

13

15

16

15

Figures

Figure 1: GC/MS chromatogram spectrum of M. longifolia oil extract.