Table of Contents  
ORIGINAL ARTICLE
Year : 2015  |  Volume : 14  |  Issue : 1  |  Page : 65-68

The composition of the lipoidal matter of the seeds of Pleiogynium timorense (DC.) Leenh


1 Department of Pharmacognosy, National Research Centre, Cairo, Egypt
2 Department of Pharmacognosy, Faculty of Pharmacy, Cairo University, Giza, Egypt
3 Chemistry of Medicinal Plants, National Research Centre, Cairo, Egypt

Date of Submission22-Dec-2014
Date of Acceptance09-Feb-2015
Date of Web Publication8-Apr-2015

Correspondence Address:
Gehan F Abdel Raoof
Department of Pharmacognosy, National Research Centre, Dokki, Cairo
Egypt
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1687-4315.154725

Rights and Permissions
  Abstract 

Background and objectives
Nothing dealing with the chemistry of the seeds of Pleiogynium timorense (DC.) Leenh. could be traced in the available literature. The objective of the present work was to investigate the lipoidal matter from P. timorense seeds and to isolate and identify the major sterols and triterpenes in the petroleum ether extract.
Materials and methods
The petroleum ether extract of P. timorense seeds was prepared. The extract was subjected to column chromatography using silica gel (type 60-230 mesh) as an adsorbent. The unsaponifiable matter and fatty acid methyl esters were analyzed by GC/MS.
Results and conclusion
GC/MS analysis of the unsaponifiable matter from the petroleum ether extract of P. timorense seeds revealed the identification of 70.04% of unoxygenated compounds and 23.34% oxygenated compounds. Hydroxylated compounds, ketones, and steroidal compounds represent 23.24, 0.20, and 3.19%, respectively. 1-Heptene (66.47%) was the major compound in the USM, followed by butylated hydroxy toluene (21.07%). GC/MS analysis of fatty acid methyl esters showed that nine fatty acid methyl ester derivatives could be identified, representing 95.84% of the total composition. The major fatty acid was 9,12-octadecadienoic acid (linoleic acid) (33.8%). Saturated fatty acids represent 36.93% of the total fatty acid content, whereas monounsaturated and diunsaturated fatty acids represent 25.11 and 33.8% of the total fatty acid content, respectively. Column chromatography of the petroleum ether extract led to the isolation of two compounds [α-amyrin and 5,24(28)-cholestadien-24-methylen-3β-ol].

Keywords: α-amyrin, 5,24(28)-cholestadien-24-methylen-3β-ol, lipids, Pleiogynium timorense, sterols


How to cite this article:
Said AA, Aboutabl EA, Hussein AA, Abdel Raoof GF. The composition of the lipoidal matter of the seeds of Pleiogynium timorense (DC.) Leenh. Egypt Pharmaceut J 2015;14:65-8

How to cite this URL:
Said AA, Aboutabl EA, Hussein AA, Abdel Raoof GF. The composition of the lipoidal matter of the seeds of Pleiogynium timorense (DC.) Leenh. Egypt Pharmaceut J [serial online] 2015 [cited 2020 Sep 19];14:65-8. Available from: http://www.epj.eg.net/text.asp?2015/14/1/65/154725


  Introduction Top


Pleiogynium timorense (DC.) Leenh. (Anacardiaceae) is an evergreen tree indigenous to tropical and subtropical regions. It is known in Arabic as Gambozia and cultivated in Egypt as an ornamental plant [1],[2]. P. timorense has edible fruits used in the preparation of jellies, jams, and preserves [3],[4]. Previous phytochemical studies resulted in the isolation of quercetin, myricetin, rutin, quercitrin, hyperin, lupeol, and β-sitosterol from the leaves. Aqueous and alcoholic extracts of the leaves showed significant antimicrobial activity against Staphylococcus aureus and Bacillus subtilis [5]. The ethanolic extract of the leaves showed significant hypoglycemic, antioxidant, and anti-inflammatory properties. In addition, 12 phenolic compounds were isolated from the leaves of the plant, which include kaempferol, gallic acid, kaempferol-3-O-β-d-galactopyranoside, kaempferol-3-O-β-d-glucopyranoside, quercetin-3-O-β-d-galactopyranoside, quercetin-3-O-β-d-glucopyranoside, kaempferol-3-O-β-d-6½-methylglucuronopyranoside, kaempferol-3-O -β-d-glucuronopyranoside, myricetin-3-O-α-l-rhamnopyranoside, 3,5-di-O-galloylquinic acid, 1, 4, 6-tri-O-galloyl-β-d-glucopyranose, and 1, 3, 4, 6-tetra-O-galloyl-β-d-glucopyranose [6]. Cyanidin-3-glucoside was isolated from the fruits [7], which is reported to have antioxidant activity. The present study was undertaken to investigate the lipoidal matter composition of P. timorense seeds.


  Materials and methods Top


Plant material

Fruits of P. timorense were collected from Zoo Garden, Giza, Egypt, in April 2010, and were kindly authenticated by Dr. Tereez Labib, Consultant of Plant Taxonomy at the Ministry of Agriculture and director of Orman Botanical Garden, Giza, Egypt, and confirmed by the taxonomist, Dr. M. El-Gebaly, NRC. The fruits were air-dried, and the seeds and the pericarp were separated, powdered, and stored in dark well-closed containers.

Preparation of the lipoidal matter

The powder of the air-dried seeds of P. timorense (800 g) was exhaustively extracted with light petroleum (60°-80°C) in a continuous extraction apparatus. The extract was evaporated under vacuum to yield 28 g of dry residue, representing 3.5% of the air-dried seeds.

Investigation of the lipoidal matter

Saponification of the petroleum ether extract

The petroleum ether extract (1 g) was subjected to saponification according to the method reported by Tsuda et al. [8]. Percentages of the unsaponifiable matter and the total fatty acid were found to be 38 and 60%, respectively.

Preparation of fatty acid methyl esters

Free fatty acids obtained by saponification were methylated according to the method reported by Finar [9].

GC/MS analysis

Both the unsaponifiable and the saponifiable fractions were studied to identify their contents using GC/MS analysis. The constituents were identified by comparison of their mass spectral fragmentation patterns with those of the available database libraries, Wiley (Wiley International, Colorado, USA) and NIST (Nat. Inst. St Technol., Colorado, USA), and/or published data [10],[11]. Quantitative determination was carried out on the basis of the peak area integration. GC/MS analysis of the unsaponifiable matter and fatty acid methyl esters of P. timorense seeds was carried out using conditions described in [Table 1].
Table 1: Conditions of GC/MS an alysis of the unsaponifiable matter and fatty acid methylesters

Click here to view


The Experiment

An NMR Jeol ECA spectrometer (JEOL Corporation, Tokyo, Japan), 500 MHz for 1 H-NMR and 125 MHz for 13 C-NMR, using CDCl 3 as a solvent, was used. All chemical shifts (δ) are given in ppm units with reference to TMS as the internal standard, and the coupling constants (J) are given in Hz. The gas chromatograph was coupled with a mass spectrometer [an Agilent 6890 gas chromatograph coupled with an Agilent mass spectrometric detector, 70 eV (Agilent Technologies, Santa Clara, California, USA), ESR, Brucker, Elexsys, E 500 (Frankfurt, Germany)].

Extraction and isolation of major sterols and triterpenes

Air-dried powdered seeds of P. timorense (800 g) were exhaustively extracted with light petroleum (60°-80°C) in a continuous extraction apparatus. The extract was evaporated under vacuum to yield 28 g of extract, representing 3.5% of the air-dried powder. Part of the extract (20 g) was subjected to column chromatography using silica gel (type 60-230 mesh, 200 g; E. Merck, Darmstadt, Germany) as an adsorbent, and elution was carried out with n-hexane, followed by n-hexane-ethyl acetate mixtures of increasing polarity. Fractions (50 ml each) were collected and concentrated separately to a small volume. All fractions were screened by thin-layer chromatography on silica gel 60 F 254 plates using hexane: ethyl acetate (9 : 1) as the solvent system and P-anisaldehyde-sulphuric acid spray reagent, followed by heating at 110°C for 5 min. Similar fractions were pooled to form two major groups and the solvents were evaporated separately under reduced pressure.

Group 1

It contained one major spot (Rf = 0.38) of violet color with the P-anisaldehyde-sulphuric acid spray reagent. The fraction was freed of the solvent under reduced pressure; the residue was dissolved in the least amount of chloroform and subjected to purification by rechromatography on preparative thin-layer chromatography using hexane: ethyl acetate (8.5 : 1.5). The bands were eluted with chloroform. The chloroformic eluate was filtered and concentrated to a small volume to yield compound L 1 (5 mg).

Group 2

It contained one major spot (Rf = 0.3) of violet color, with the P-anisaldehyde-sulphuric acid spray reagent. The fraction was freed of the solvent under reduced pressure. The residue obtained was purified as in group I to yield compound L 2 (10 mg).


  Results and discussion Top


A total of 28 compounds were identified [Table 2] by GC/MS of the unsaponifiable matter, representing 96.68% of the total unsaponifiable content. The analysis revealed that the identified components consisted of 70.04% unoxygenated compounds and 23.34% oxygenated compounds: 23.24% hydroxylated compounds, 0.10% ketones, and 3.19% steroids. 1-Heptene (66.47%) was the major compound in the USM, followed by butylated hydroxy toluene (21.07%). About nine fatty acid methyl ester derivatives could be identified [Table 3], representing 95.84% of the total composition. The major fatty acids were 9,12-octadecadienoic acid (linoleic acid) (33.8%), followed by 9-octadecenoic acid (oleic acid) (24.14%) and 14-methyl-pentadecanoic acid 12.08%. Saturated fatty acids represent 36.93% of the total fatty acid content, whereas monounsaturated and diunsaturated fatty acids represent 25.11 and 33.8% of the total fatty acid content, respectively.
Table 2: GC/MS analysis of the unsaponifiable matter from Pleiogynium timorense seeds

Click here to view
Table 3: GC/MS analysis of fatty acid methylesters from Pleiogynium timorense seeds

Click here to view


Compound L 1 : white powder (5 mg), m.p. (181°-183°C), a single spot in hexane : ethyl acetate (8.5 : 1.5) (Rf = 0.38), violet color with the P-anisaldehyde-sulphuric acid spray reagent.

Mass spectral data: 426(M + ), 411 (M + -CH 3 ), 409 (M +1 -H 2 O), 218 (100%) 203 (28%) and 189 (29%). The mass spectrum showed a molecular ion peak at m/z 426 for the molecular formula C 30 H 50 O, in addition to the following peaks at m/z 411(M + -CH 3 ), 409 (M +1 -H 2 O), 218 (M-208 (C 14 H 24 O), and 203 (218-CH 3 )+. On the basis of the spectral data, melting point, and comparison with an authentic sample, compound L 1 was identified as α-amyrin. This compound was previously isolated from P. timorense leaves [5]. This is the first report in the seeds of the plant.

Compound L 2 : white powder (10 mg), m.p. (142°-143°C), gave a single spot in hexane : ethyl acetate (8.5 : 1.5) (Rf = 0.3), gave violet color with the P-anisaldehyde-sulphuric acid spray reagent.

1 H-NMR spectral data of compound L 2 1 H-NMR, CDCl 3 : δ 5.35(H-6, broad s), δ 3.9(H-3, m), δ 4.6, δ 4.5 (H-28, s), δ 0.82(3H-18, s), δ 0.84(3H-19, s), δ 1.4 (3H-26,d, J=7 Hz), δ 1.4(3H-27,d, J = 7 Hz), δ 0.9 (3H-21,d, J=6.5 Hz), δ 2.2(H-25, m)

13 C-NMR in CDCl 3 spectral data of compound L 2 : The spectrum showed peaks at δ 140.813 (C-5), δ 147.5 (C-24), 129.7 (C-6), 111.3 (C-28), 67 (C-3), 19.3 (C-21), 11.8 (C-19,C18), 22.6 (C-26, C27), 30.9 (C-23), 33.8 (C-25), 34.5 (C-22), 35.6 (C-20). Thus, compound L 2 is identified as 5,24 (28)-cholestadien-24-methylen-3β-ol, which was isolated, for the first time, from the plant.


  Conclusion Top


This study showed the identification of 28 compounds of unsaponifiable matter from the seeds of P. timorense, which were identified by GC/MS. 1-Heptene (66.47%) was the major compound in the USM. About nine fatty acid methyl ester derivatives could be identified. The major fatty acid was 9,12-octadecadienoic acid (linoleic acid) (33.8%).


  Acknowledgements Top


The authors thank the National Research Centre, Egypt, for funding this work.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Bailey LH. The standard encyclopedia of horticulture. New York: The Mac Millan Company; 1953. III:2713.  Back to cited text no. 1
    
2.
Jessup LW. Pleiogynium. Flora of Australia: Australia; 1985. 25.  Back to cited text no. 2
    
3.
Winton AL, Winton KB. The structure and composition of foods. New York: John Wiley and Sons, Inc.; 1935. II:728.  Back to cited text no. 3
    
4.
Everett TH. The New York botanical garden illustrated encyclopedia of horticulture. New York, London: Carland publishing Inc. 1981. 8:2721.  Back to cited text no. 4
    
5.
El-Fiki NM, Ahmed FI. Phytochemical study of Pleiogynium solandri (Benth.) Engl. Azhar J Pharm Sci 1999; 24:38-50.  Back to cited text no. 5
    
6.
Al Sayed E, Martiskainen O, Sinkkonen J, Pihiaja K, Ayoub N, Singab AE, El-Azizi M. Chemical composition and bioactivitiy of Pliogynium timorense (Anacardiaceae). Nat Prod Commun 2010; 5:545-550.  Back to cited text no. 6
    
7.
Netzel M, Netzel G, Tian Q, Schwartz S, Konczak I. Native Australian fruits - a novel source of antioxidants for food. Innov Food Sci Emerg 2007; 8:339-346.  Back to cited text no. 7
    
8.
Tsuda K, Sakai K, Tanabe K, Kishida Y. Isolation of 22-dehydrocholesterol from Hypnea japonica. J Am Chem Soc 1960; 82:1442-1443.  Back to cited text no. 8
    
9.
Finar IL. Organic chemistry. 5th ed. London, UK: Longmans Green and Co. Ltd; 1967. 1:212.  Back to cited text no. 9
    
10.
Adams RP. Identification of essential oils by ion trap mass spectroscopy. New York: Academic Press Inc.; 1995.  Back to cited text no. 10
    
11.
Jennings W, Shibamato T. Qualitative analysis of flavor and fragrance volatiles by glass capillary gas chromatography. New York: Academic Press; 1981.  Back to cited text no. 11
    



 
 
    Tables

  [Table 1], [Table 2], [Table 3]


This article has been cited by
1 Phytoconstituents and bioactivities of the Bark of Pleiogynium timorense (DC.) Leenh (Anacardiaceae)
Gehan Fawzy Abdel Raoof,Ataa Abdelhaleem Said,Khaled Younes Mohamed,Hesham A. Gomaa
Journal of Herbmed Pharmacology. 2020; 9(1): 20
[Pubmed] | [DOI]



 

Top
 
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Introduction
Materials and me...
Results and disc...
Conclusion
Acknowledgements
References
Article Tables

 Article Access Statistics
    Viewed1425    
    Printed44    
    Emailed0    
    PDF Downloaded140    
    Comments [Add]    
    Cited by others 1    

Recommend this journal


[TAG2]
[TAG3]
[TAG4]