alexa
Reach Us +441414719275
Characterization of the Leaf Essential Oil Composition of Annona squamosa L. from Foothills of North India | OMICS International
ISSN: 2167-0412
Medicinal & Aromatic Plants
Make the best use of Scientific Research and information from our 700+ peer reviewed, Open Access Journals that operates with the help of 50,000+ Editorial Board Members and esteemed reviewers and 1000+ Scientific associations in Medical, Clinical, Pharmaceutical, Engineering, Technology and Management Fields.

Characterization of the Leaf Essential Oil Composition of Annona squamosa L. from Foothills of North India

Ram Swaroop Verma1*, Neeta Joshi2, Rajendra Chandra Padalia1, Ved Ram Singh3, Prakash Goswami1 and Amit Chauhan1

1CSIR - Central Institute of Medicinal and Aromatic Plants, Research Centre Pantnagar, Udham Singh Nagar, Uttarakhand, India

2Department of Chemistry, MB Government PG College Haldwani, Kumaun University, Nainital, Uttarakhand, India

3CSIR - Central Institute of Medicinal and Aromatic Plants, Lucknow, Uttar Pradesh, India

*Corresponding Author:
Ram Swaroop Verma
CSIR - Central Institute of Medicinal and Aromatic Plants Research
Centre Pantnagar, Udham Singh Nagar, Uttarakhand, India
Tel: +919756021222
E-mail:
[email protected]

Received date: October 14, 2016; Accepted date: October 19, 2016; Published date: October 25, 2016

Citation: Verma RS, Joshi N, Padalia RC, Singh VR, Goswami P, et al. (2016) Characterization of the Leaf Essential Oil Composition of Annona squamosa L. from Foothills of North India. Med Aromat Plants (Los Angel) 5:270. doi: 10.4172/2167-0412.1000270

Copyright: © 2016 Verma RS, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Visit for more related articles at Medicinal & Aromatic Plants

Abstract

The leaf essential oil composition of Annona squamosa L., collected from the lower region of Himalaya was investigated using gas chromatography-flame ionisation detector (GC-FID) and GC-mass spectrometry (GC-MS). A total of forty-three constituents, representing 88.6% of the total oil composition were identified. The essential oil was primarily composed of sesquiterpenoids (sesquiterpene hydrocarbons: 63.4% and oxygenated sesquiterpenes: 21.8%). Major constituents of the oil were (E)-caryophyllene (15.9%), γ-cadinene (11.2%), epi-α-cadinol (9.4%), (Z)- caryophyllene (7.3%), γ-muurolene (5.4%), α-humulene (5.2%), viridiflorene (5.0%), α-cadinol (3.9%), aromadendrene (2.9%), δ-cadinene (2.9%), α-cadinene (2.9%), (2Z,6Z)-farnesal (2.2%) and caryophyllene oxide (2.1%).

Keywords

Annona squamosa; Annonaceae; Essential oil composition; Sesquiterpenes

Introduction

Annona L. (Annonaceae) comprises approximately 162 species of trees and shrubs that are found predominantly in lowland tropical regions [1]. Annona squamosa L., commonly known as a ‘custard apple’ or ‘sharifa’ is native to South America and the West Indies, but now it is cultivated throughout India for its nutritive fruits [2]. In Indian system of medicine, it is used as antitumour, diuretic and for wound healing. The leaves are anthelmintic and the powdered unripe fruits are taken internally in the form of paste with water for the treatment of diarrhoea and dysentery [3]. The plant is used in traditional medicines of several tropical countries to treat epilepsy, dysentery, cardiac problems, worm infection, constipation, haemorrhage, antibacterial, dysurea, fever and ulcer. The young leaves of the plant are used extensively for their antidiabetic activity [4]. Seeds of the plant are well known for killing head lice [5]. The dried unripe fruit powder is used to destroy vermin. A paste of seed powder has been used for killing worm in the wound of cattles [6]. The plant is also attributed with antifertility, abortifacient, antitumor and antimalarial activities [7-9]. An alkaloid, p-hydroxy benzyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, isolated from the leaves is reported to be act as cardiotonic [10]. A fraction of total root alkaloids is reported to be antihypertensive, antispasmodic, antihistaminic and a bronchodilator [11]. The leaf essential oil of the plant has been demonstrated potent trypanocidal and antimalarial activities [12].

The chemical composition of the essential oil of A. squamosa has been studied previously from different countries. It is evident from the literature that oil of A. squamosa is exist in six different chemotypic forms which are: (i) monoterpene hydrocarbon rich; (ii) oxygenated monoterpene rich; (iii) sesquiterpene hydrocarbon rich; (iv) oxygenated sesquiterpene rich; (v) oils with relative amounts of sesquiterpene hydrocarbons and oxygenated sesquiterpenes and (vi) oil containing sesquiterpene and diterpenoids compounds [13]. The leaf essential oil from Benin possessed isocaryophyllene, camphene, β-caryophyllene, epi-α-cadinol and epi-α-muurolol as major constituents [14]. The leaf essential oil rich in (E)-caryophyllene, germacrene D, bicyclogermacrene, (Z)-caryophyllene, β-elemene and α-humulene is reported from Brazil [12]. Major constituents of the leaf essential oil of A. squamosa from north Indian plains are (E)-caryophyllene, germacrene D, bicyclogermacrene, β-elemene, γ-cadinene, α-muurolol and aromadendrene [15]. The leaf oil from southern India is dominated by β-cedrene and β-caryophyllene [16]. The root essential oil of A. squamosa rich in β-caryophyllene, α-pinene, α-humulene and α-gurjunene is reported from Brazil [17]. Main components of the leaf oil from France are germacrene D, β-elemene, α- and β-pinene, sabinene, bicyclogermacrene, τ-cadinol and τ-muurolol, while the fruit oil contained spathulenol, bornyl acetate, germacrene D, borneol and verbenone [18]. The fruit oil contained τ-cadinol, τ-muurolol, spathulenol, α-copaene and α-terpineol is reported from Malaysia [19]. Moreover, the bark essential oil contain 1H-cycloprop(e)azulene, germacrene D, bisabolene, caryophyllene oxide, bisabolene epoxide and kaur-16-ene as main constituents [20].

A review of literature revealed that there is no report on the essential oil composition of A. squamosa growing in the foothill region of north India. Therefore, in this research, leaf essential oil composition of A. squamosa collected from foothills of Uttarakhand has been investigated using gas chromatography and gas chromatography-mass spectrometry.

Materials and Methods

Plant material and isolation of essential oil

Fresh leaves of A. squamosa were collected from the experimental field of CSIR-Central Institute of Medicinal and Aromatic Plants, Research Centre, Pantnagar, Uttarakhand in the month of November (Figure 1). The plant material was authenticated at the Botany Department, CIMAP Research Centre Pantnagar by one of the authors (AC). The experimental site is located between coordinates 29.02° N, 79.31° E and an altitude of 243 m in foothills of north India. Isolation of the essential oil from shade dried leaves was carried out by hydrodistillation for 3 h using a Clevenger’s type apparatus. Isolated oil was dried over anhydrous Na2SO4 and stored at 4°C until further analyses.

Figure 1: Twigs of Annona squamosa showing leaf, flower and fruit.

Gas chromatography (GC/FID)

Gas chromatography-flame ionization detection (GC-FID) was performed for quantification of the essential oil constituents. GC analysis of the essential oil was carried out on a PerkinElmer AutoSystem XL gas chromatograph, equipped with DB-5 capillary column (60 m × 0.32 mm i.d., film thickness 0.25 μm) and flame ionization detector (FID). The oven column temperature ranged from 70-250°C, programmed at 3°C min-1, with initial and final hold time of 2.0 min, using H2 as carrier gas at 10 psi constant pressure, a split ratio of 1:35, an injection size of 0.03 μL neat, and injector and detector temperatures were maintained at 250°C and 280°C, respectively.

Gas chromatography-Mass spectrometry (GC/MS)

Gas chromatography-mass spectrometry (GC-MS) was performed for identification of the essential oil constituents. GC-MS analysis of the essential oil was carried out on a Clarus 680 GC interfaced with a Clarus SQ 8C mass spectrometer of PerkinElmer fitted with Elite-5 MS fused-silica capillary column (5% phenyl polysiloxane, 30 m × 0.25 mm internal diameter, film thickness 0.25 μm). The oven temperature program was from 60°C to 240°C, at 3°C min-1, and programmed to 270°C at 5°C min-1. Injector temperature was 250°C; transfer line and source temperatures were 220°C; injection size 0.03 μL neat; split ratio 1:50; carrier gas He at 1.0 mL min-1; ionization energy 70 eV; mass scan range 40-450 amu.

Identification of essential oil constituents

Characterization of the essential oil constituents was carried out on the basis of retention index (RI), calculated using a homologous series of n-alkanes (C8-C30, Supelco Bellefonte, PA, USA) under identical experimental conditions; mass spectra library search (NIST/EPA/NIH, version 2.0 g, and Wiley registry of mass spectral data 9th edition) and by comparing the mass spectral and retention data with literature [21]. The relative amounts of individual components were calculated based on the GC peak area (FID response) without using a correction factor.

Statistical analysis

To examine whether the essential oil constituents identified are useful in reflecting the chemical relationships between different compositions, their contents (%) were subjected to hierarchical cluster analysis using average method [22]. This software computes the hierarchical clustering of a multivariate dataset based on dissimilarities. The derived dendrogram depicts the grouping of chemical compositions as per their chemical constituents.

Results and Discussion

The shade dried leaves of A. squamosa yielded 0.13% essential oil on hydrodistillation. The resulting essential oil was subjected to GC-FID and GC-MS analyses. Altogether, forty-three constituents, representing 88.6% of the total oil composition were identified. The detailed results are summarised in Table 1. The essential oil composition was mainly dominated by sesquiterpenoids (sesquiterpene hydrocarbons: 63.4% and oxygenated sesquiterpenes: 21.8%) (Figure 2). Major sesquiterpenoid constituents of the oil were (E)-caryophyllene (15.9%), γ-cadinene (11.2%), epi-α-cadinol (9.4%), (Z)-caryophyllene (7.3%), γ-muurolene (5.4%), α-humulene (5.2%), viridiflorene (5.0%), α-cadinol (3.9%), aromadendrene (2.9%), δ-cadinene (2.9%), α-cadinene (2.9%), (2Z,6Z)-farnesal (2.2%), caryophyllene oxide (2.1%), spathulenol (1.9%), β-elemene (1.9%), humulene epoxide II (1.8%), δ-elemene (1.5%) and α-copaene (1.3%). However, notable monoterpenoids constituents of the oil were p-cymene (0.8%), limonene (0.8%) and bornyl acetate (0.5%). The structures of the major constituents of the oil are presented in Figure 3.

S. No. Compound RIa RIb Content (%)
1. α-Pinene 927 932 0.1
2. Myrcene 983 988 0.1
3. α-Phellandrene 996 1002 t
4. p-Cymene 1018 1020 0.8
5. Limonene 1022 1024 0.8
6. 1,8-Cineole 1024 1026 t
7. (Z)-β-Ocimene 1028 1032 t
8. (E)-β-Ocimene 1040 1044 0.1
9. γ-Terpinene 1051 1054 0.1
10. Terpinolene 1082 1086 t
11. Linalool 1094 1095 0.2
12. Camphor 1137 1141 0.3
13. Terpinen-4-ol 1171 1174 0.2
14. Myrtenol 1191 1194 0.2
15. Bornyl acetate 1279 1284 0.5
16. δ-Elemene 1331 1335 1.5
17. Cyclosativene 1364 1369 t
18. α-Copaene 1369 1374 1.3
19. β-Bourbonene 1382 1387 t
20. β-Elemene 1385 1389 1.9
21. (Z)-Caryophyllene 1402 1408 7.3
22. (E)-Caryophyllene 1412 1417 15.9
23. Aromadendrene 1434 1439 2.9
24. α-Humulene 1448 1452 5.2
25. γ-Muurolene 1472 1478 5.4
26. Germacrene D 1474 1480 t
27. β-Selinene 1484 1489 t
28. Viridiflorene 1490 1496 5.0
29. α-Selinene 1492 1498 t
30. α-Muurolene 1494 1500 t
31. γ-Cadinene 1508 1513 11.2
32. δ-Cadinene 1516 1522 2.9
33. trans-Calamenene 1518 1521 t
34. α-Cadinene 1532 1537 2.9
35. α-Calacorene 1538 1544 t
36. Elemol 1542 1548 t
37. Spathulenol 1572 1577 1.9
38. Caryophyllene oxide 1576 1582 2.1
39. Globulol 1584 1590 0.5
40. Humulene epoxide II 1604 1608 1.8
41. epi-α-Cadinol 1634 1638 9.4
42. α-Cadinol 1648 1652 3.9
43. (2Z,6Z)-Farnesal 1676 1683 2.2
44. Class composition      
45. Monoterpene hydrocarbons     2.0
46. Oxygenated monoterpenes     1.4
47. Sesquiterpene hydrocarbons     63.4
48. Oxygenated sesquiterpenes     21.8
49. Total identified (%)     88.6

Table 1: Leaf essential oil composition of Annona squamosa from Uttarakhand, India.

Figure 2: Class composition of the leaf essential oil of Annona squamosa collected from foothills of north India.

Figure 3: Structures of major constituents of Annona squamosa leaf essential oil.

The leaf essential oil composition of A. squamosa has been investigated earlier from different countries (Table 2). Major constituents of the Brazilian oil were (E)-caryophyllene (27.4%), germacrene D (17.1%), bicyclogermacrene (10.8%), (Z)-caryophyllene (7.3%), β-elemene (6.2%) and α-humulene (5.7%) [12]. Two distinct compositions were described from Egypt. First one was dominated by carvone (24.9%), diacetyl (9.3%) and linalool (7.7%) [23], while other composition was characterised by higher amount of β-gurjunene (42.49%), viridiflorene (6.68%), aromadendrene (5.49%), γ-muurolene (5.72%) and allo-aromadendrene epoxide (5.31%) [24]. Moreover, germacrene D (15.7%), β-elemene (12.0%), sabinene (8.8%) and α-pinene/β-pinene (8.1%) were major constituents of the leaf oil from France [18]. Further, major constituents of the leaf oil from north Indian plains were (E)-caryophyllene (22.9%), germacrene D (21.3%), bicyclogermacrene (8.5%), β-elemene (7.8%), γ-cadinene (6.7%) and α-muurolol (5.7%) [15]. On the other hand, leaf oil from southern India was characterised by higher amounts of β-cedrene (23.3%), β-caryophyllene (14.1%), (E,E)-farnesol (7.0%) and cadina-1,4-diene (6.9%) [16].

County Major Constituents Reference
Brazil (E)-Caryophyllene (27.4%), germacrene D (17.1%), bicyclogermacrene (10.8%), (Z)-caryophyllene (7.3%), β-elemene (6.2%), α-humulene (5.7%) [12]
Egypt β-Gurjunene (42.49), viridiflorene (6.68%), aromadendrene (5.49%), γ-muurolene (5.72%), allo-aromadendrene epoxide (5.31%) [24]
Egypt Carvone (24.9%), diacetyl (9.3%), linalool (7.7%) [23]
India (E)-Caryophyllene (22.9%), germacrene D (21.3%), bicyclogermacrene (8.5%), β-elemene (7.8%), γ-cadinene (6.7%), α-muurolol (5.7%) [15]
India β-Cedrene (23.3%), β-caryophyllene (14.1%), (E,E)-farnesol (7%), cadina-1,4-diene (6.9%), allo-aromadendrene (5.5%), calamenene (5.1%) [16]
France Germacrene D (15.7%), β-elemene (12.0%), sabinene (8.8%), bicyclogermacrene (6.0%), τ-cadinol (5.5%) [18]

Table 2: Leaf essential oil composition of Annona squamosa reported from different counties.

To compare the composition of the investigated A. squamosa essential oil with earlier reported oil compositions [12,15,16,18,23,24], twenty-four selected components (amount ≥ 5.0%) of different oils were subjected to the hierarchical cluster analysis. The derived dendrogram clearly showed dissimilarity based on the percentages of the constituents present in the different oil compositions. The examined oil from foothill region of Uttarakhand was quite different from the oils of other regions, hence made a separate group in the hierarchical cluster analysis (Figure 4).

Figure 4: Hierarchical cluster analysis of the leaf essential oil composition of Annona squamosa. 1: Foothills, north India (present study); 2: north Indian plain [15]; 3: southern India [16]; 4: Brazil [12]; 5: Egypt [24]; 6: France [18]; 7: Egypt [23]. For detail compositions of the oils see Tables 1 and 2.

Though the examined essential oil was rich in sesquiterpenoid compounds, there were substantial quantitative and qualitative variations when compared with previous reports from India and other parts of the world. Noteworthy observation is the fact that compounds such as carvone, diacetyl, cadina-1,4-diene, β-cedrene, β-gurjunene, bicyclogermacrene, which were the major constituents in previous studies [15,16,18,23,24] could not be detected in this investigation. Moreover, germacrene D, a major constituent of various previous studies on A. squamosa leaf oil [12,15,18] was present in only trace amount in the examined oil. This may be attributed to several factors such as climatic condition, season, and age of the plant, genotype, and processing procedures.

Conclusions

In this study, leaf essential oil composition of A. squamosa was analysed from foothills of north India. The essential oil was dominated by sesquiterpenoids (85.2%) with (E)-caryophyllene (15.9%), γ-cadinene (11.2%), epi-α-cadinol (9.4%), (Z)-caryophyllene (7.3%), γ-muurolene (5.4%), α-humulene (5.2%) and viridiflorene (5.0%) as major constituents. The examined oil showed considerable dissimilarity in chemical composition with previously reported leaf essential oil compositions from other regions.

Acknowledgements

Authors are thankful to Director, CSIR-CIMAP Lucknow for continuous encouragement and providing necessary facilities to carry out the work. Authors are also thankful to the Central Chemical Facility, CIMAP Lucknow for providing facility for GC and GC-MS analysis.

References

Select your language of interest to view the total content in your interested language
Post your comment

Share This Article

Relevant Topics

Article Usage

  • Total views: 10381
  • [From(publication date):
    October-2016 - Oct 20, 2018]
  • Breakdown by view type
  • HTML page views : 10240
  • PDF downloads : 141
 

Post your comment

captcha   Reload  Can't read the image? click here to refresh

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2018-19
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001Extn: 9040

Clinical Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

Food & Nutrition Journals

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

General Science

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics & Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Materials Science Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Nursing & Health Care Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

Ann Jose

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

https://www.gaziantepescort.info

Steve Harry

[email protected]

1-702-714-7001Extn: 9042

 
© 2008- 2018 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version