GC/MS Analysis of the Peels of Annona muricata L.

 

Widad M.K. Al-ani1*, Sarah S. Almahdawy2*

1Department of Pharmacy, Ashur University College, Baghdad, Iraq.

2Department of Pharmacy, Al-Mustafa University College.

*Corresponding Author E-mail: wmkalani@gmail.com, Sara.saad.abd @gmail.com

 

ABSTRACT:

This research is designed to identify the chemical composition of crude drug of the peel of Annona muricata L. by placing the peels powder in Soxhlet apparatus. 95% of hexane was put in the round bottom flask attached to this setup. Then the whole setup was placed on the heating mantle. The temperature was setup to 60-80C. The obtained results show the presence of 18 compounds. The most important constituents are: humulen (10.68%), longipinane (2.09%), cyclononasiloxane (1.22%), then followed by borneol acetate represents about (0.93%), alpha limonene diepoxide (0.87%), copaene (0.62), alpha pinene (0.56%), caryophylline oxide (0.55%), Dodecamethylcyclohexasiloxane (0.52%), kaurene (0.52%), retinal (0.46%) while the minor constituents are Germacrene, beta pinene, aromadendrene, and 4,7-Octadecadienoic acid, methyl ester is the minimum one. Due to good result obtained from the crude peel extract of this plant so further studies chemically and clinically of the active compounds is required.

 

KEYWORDS: Graviola; Annona muricata L.; Gas Chromatography/Mass Spectrometer.

 

 


INTRODUCTION:

Throughout the world, either developing or under-developing countries, plants have been used as medicine to meet primary healthcare needs1. It is estimated that vast majority of population depends on medicinal plants for therapy against several diseases or disorders.2

 

Many forms of alternative medicines are available for those who do not want regular medicine (or) who cannot be helped by conventional medicine, among these important medicinal plants which have many medicinal properties is Annona muricata L.3,4

 

It is a small evergreen tree 5-6 meter height with large, glossy, green leaves. It produce a large heart shaped, edible fruit about 5-20cm in diameter, yellow greenish in colour and has white flesh inside.5

 

Annonaceae family which is commonly known as Graviola or Soursop or Gunbanana, the name soursop is due to sour and sweet flavour of its fruit.

 

A.muricata is a tropical fruit-bearing tree belong to The soursop is native to tropical Central and South America and the Caribbean, but is now widely cultivated in tropical areas worldwide, including southern Florida and Southeast Asia, from sea level to altitudes of around 1150 meters.6

 

The phytochemicals present in A. muricata are alkaloids, Flavonoids, carbohydrates, cardiac glycosides, tannins, saponins, phytosterol, terpenoids and proteins. Several isoquinoline alkaloids: reticuline, coclaurine, coreximine, atherosperminine, stepharine, and anomuricine have been isolated from the roots, leaves and stem barks of A. muricata. The essential oil of the fresh fruit pulp of A. muricata was found to contain 2-hexenoic acid methyl ester (23.9%), 2-hexenoic acid ethyl ester (8.6%), 2-octenoic acid methyl ester (5.4%), 2-butenoic acid methyl ester (2.4%), β-caryophyllene (12.7%), 1,8-cineole (9.9%), linalool (7.8%), and calarene (2.2%).7

 

The major pharmacological activities of the plant includes anticarcinogenic, genotoxic, wound healing and antimicrobial activity, that it possess a potent anticancerous agents coined as Acetogenins (figure 1), which play important role towards many types of cancer, Acetogenins are potent inhibitors of NADH oxidase (nicotinamide adenine dinucleotide phosphate-oxidase) of the plasma membranes of cancer cells therefore its fruit has an economic worth and hence cultivated and used widely as an edible food.8

 

Figure 1: Structure of acetogenin

 

MATERIALS AND METHODS:

Plant collection:

The plant peels of A. muricata were collected from local market in Iraq. The peels of the plant were air dried and grounded by a miller (figure 2).

 

Figure 2: Annona muricata L.

 

Extraction procedure:

Aqueous and alcoholic extract of the sample was prepared by soaking 5gm of dried powder in 100ml of water, ethanol respectively for 12hrs. The extract was filtered using the Whatmann filter paper and it was used for further studies.8

 

50gm of A.muricata peels powder was placed in to Soxhlet apparatus. 95% of hexane was taken in the round bottom flask attached to this setup. Then the whole setup was placed on the heating mantle. The temperature was setup to 60-80C the hexane gets vaporized and rises up to the condenser where it condenses back in to the liquid and falls in to the plant sample in the cone and extract certain compounds falls in to the round bottom flask.9

 

Phytochemical screening procedure:

Qualitative tests for alkaloids, flavonoids, glycosides, saponins, tannins, terpenoids, and anthraquinone. Mayers test, for Alkaloids, Shinodas test for flavonoids, Keller-Killani test for cardiac glycosides, Froth test for saponins, Lead acetate test for tannins, Salkowsk test for terpenoids, and Ammonia test for anthraquinone were performed.10

 

Gas chromatography/mass spectroscopy (GC/MS):

This technique is done at ministry of science and technology, the GC/MS condition was: Stationary phase: modified phase silica DB5, Column: 30m X 0.25mm Internal Diameter (I.D).Mobile phase (Carrier gas): Helium split ratio (1:15)

 

Injection volume: 0.5, Column oven temperature: 70.0C, Injection temperature: 240.00C, Injection mode: Splitless, Sampling time: 1.00 min., Flow control mode: pressure, Pressure: 100.0KPa, Total flow rate: 19.9 mL/min, column flow: 1.53 mL/min, Temperature program: from 70Co (3 min) to 240 Co (8 min).

 

RESULTS AND DISCUSSION:

The result of phytochemical analysis is shown in table (1), preliminary phytochemical screening of plant peels extract appeared the presence of alkaloids, flavonoids, glycosides, saponins, tannins, anthraquinone and terpenoids in the fruit peels of this plant

 

Table (1): phytochemical screening results of A.muricata plant

Chemical test

Water

Ethanol

Alkaloids

-

+

Flavonoids

+

+

Glycosides

+

+

Saponins

+

+

Tannins

+

+

Terpenoids

+

-

Anthraquinone

+

-

 

GC/MS chromatogram of the plant peel extract (figure 3) show the presence of 18 important compounds which are listed below in table (2)


 

 

Figure 3: The graphical results of the GC/ MS analysis for the hexane extract of A. muricata peel

 

Table (2): Chemical compounds detected in A. muricata peels hexane extract by GC/MS

Name

Peak

Time

Area%

Height%

Molecular Peak

Base peak

Alpha-pinene

2

2.523

0.56

0.96

136

93

Beta-pinene

3

3.483

0.33

0.57

136

93

Pinene epoxide

5

4.967

0.25

0.46

152

109

Borneol acetate

9

7.322

0.93

1.55

196

95

Dodecamethylcyclohexasiloxane

10

7.726

0.52

0.87

444

73

Copaene

12

8.655

0.62

0.93

204

105

Germacrene D

13

8.818

0.43

0.66

204

105

3-Butoxy-1,1,1,7,7,7-hexamethyl-3,5,5tris(trimethylsiloxy)tetrasiloxane

15

10.036

0.44

0.74

590

73

Aromadendrene

16

10.497

0.29

0.29

204

105

4,7-Octadecadiynoic acid, methyl ester

17

10.964

0.24

0.38

290

105

Caryophyllene oxide

19

11.413

0.55

0.84

220

93

Retinal (Vitamin A)

22

12.621

0.46

0.65

284

109

Alpha limonene diepoxide

24

13.204

0.87

0.86

196

43

Pentadecanoic acid

28

15.492

3.54

3.72

242

73

Kaurene

31

16.683

0.52

0.76

272

91

Humulen

41

20.193

10.68

5.44

204

91

Longipinane

42

20.563

2.09

2.80

206

109

Cyclononasiloxane

53

24.221

1.22

1.07

282

57

 


The constituents of the plant extract and their percentage vary from region to region. The major constituents are humulen (10.68%), longipinane (2.09%), cyclononasiloxane (1.22%), then followed by borneol acetate represents about (0.93%), alpha limonene diepoxide (0.87%), copaene (0.62), alpha pinene (0.56%), caryophylline oxide (0.55%), Dodecamethylcyclohexasiloxane (0.52%), kaurene (0.52%), retinal (0.46%) while the minor constituents are Germacrene, beta pinene, aromadendrene, and 4,7-Octadecadienoic acid, methyl ester is the minimum one. Most of these compounds are important in our life for example: Dodecamethylcyclohexasiloxane (cyclohexasiloxane) used in cosmetic and personal care products, and can be used in dermal exposure and inhalation toxicity study.11 Alpha & beta pinene are miracle gifts of nature because they are used as anti-inflammatory agents, antimicrobial, anticancer, anti ulcerogenic, anticoagulant, antioxidant, and anticonvulsant.12 Germacrene D has antibacterial property, cytotoxicity, and antioxidant.13 Copaene used as antimicrobial, antiproliferative, antioxidant, antigenotoxic, and for cytotoxicity.14 Kaurene is so important because it is used as antimicrobial, anti-inflammatory, cardiovascular, diuretic, anti-HIV, and cytotoxic effects.15,16

 

The constituents of A.muricata peels in this study differ when compared to other region, that crude drug extract of A.muricata from India contains 4H-Pyran-4-one, 2,3-dihydro-3,5- dihydroxy-6, Tetradecanoic acid, 3,7,11,15-Tetramethyl-2-hexadecen-1-ol, Hexadecanoic acid methyl ester, nHexadecanoic acid, Phytol and Octadecanoic acid.16 while the crude drug extract of A.muricata from Uganda contains 7-Tetradecenal, n-Hexadecanoic acid, Oleyl Alcohol, Phytol, cis, cis, cis-7,10,13-Hexadecatrienal, 2-Pentadecanol, 9,12-Octadecadienoic acid, ethyl ester, 1,2-Benzenedicarboxylic acid, butyl octyl ester, and 13-Octadecatriene.17,18 and the crude drug extract of seeds of A.muricata from Nigeria contains hexadecanoic acid, 2,6-dimethyl-1,7-octadien-3-ol, 9-octadecanoic acid and nonadecanoic acid.19,20

 

CONCLUSIONS:

The extract of A. muricata peels that collected from local market in Iraq showed the presence of alkaloids, flavonoids, glycosides, saponins, tannins, terpenoids, and anthraquinone by preliminary phytochemical screening test. Then extraction of crud drug exract by soxhlet apparatus and analyzed by GC/MS identify 18 components. The major constituents are humulen (10.68%), longipinane (2.09%), cyclononasiloxane (1.22%), then followed by borneol acetate represents about (0.93%), alpha limonene diepoxide (0.87%), copaene (0.62), alpha pinene (0.56%), caryophylline oxide (0.55%), Dodecamethylcyclohexasiloxane (0.52%), kaurene (0.52%), retinal (0.46%) while the minor constituents are Germacrene, beta pinene, aromadendrene, and 4,7-Octadecadiynoic acid, methyl ester is the minimum one.

 

ACKNOWLEDGEMENT:

The author would like to acknowledge the ministry of science and technology for their help.

 

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Received on 14.09.2021 Modified on 08.12.2021

Accepted on 18.01.2022 RJPT All right reserved

Research J. Pharm. and Tech. 2022; 15(7):3137-3140.

DOI: 10.52711/0974-360X.2022.00524