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Phytochemical Composition of Methanolic Extract of Phyllanthus acidus L

(Skeels) Fresh Leaves by GC/MS Analysis

Article  in  Research Journal of Pharmacy and Technology · April 2016

DOI: 10.5958/0974-360X.2016.00106.2

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 Research J. Pharm. and Tech. 9(5): May 2016

ISSN 0974-3618 (Print) www.rjptonline.org

0974-360X (Online)

RESEARCH ARTICLE

Phytochemical Composition of Methanolic Extract of Phyllanthus acidus L

(Skeels) Fresh Leaves by GC/MS Analysis

Rohan Sharadanand Phatak1*, Anup Subhash Hendre2, Pushpa Prakash Durgawale2

1
Directorate of Research, Krishna Institute of Medical Sciences Deemed University, Malkapur, Karad-415539,
India,
2
Department of Biochemistry Krishna Institute of Medical Sciences, Malkapur, Karad-415539, India
*Corresponding Author E-mail: phatak.rohan1983@gmail.com

ABSTRACT:
Background: Secondary metabolites and different phytochemicals are present in the medicinal plant
Phyllanthus acidus belonging to family Phyllanthaceae. Scientific evidence data related GC-MS analysis of
methanolic extract of Phyllanthus acidus fresh leaves has not been reported till date during literature review.
Objective: The study was aimed to investigate phytochemicals present in the methanolic extract of Phyllanthus
acidus fresh leaves. Material and Methods: Methanolic extraction of fresh leaves Phyllanthus acidus was
prepared by simple maceration and GC-MS analysis data was provided by IICPT, Thanjavur. Phytochemical
investigation was carried out to identify the possible components from Phyllanthus acidus fresh leaves by
evaluating retention time in the GC-MS chromatogram. Results: Eleven peaks have revealed in the GC-MS
spectrum of the methanolic extract of Phyllanthus acidus. From the results, squalene is present as major
bioactive compounds in methanolic extract of Phyllanthus acidus fresh leaves. Conclusion: The present data
provides to find the molecular formula and weight of 11 biomolecules. Further investigation should be done to
isolate bioactive compounds and their structural elucidation and screening of pharmacological activity in the
drug development.

KEYWORDS: GC/MS analysis, Phytochemicals, Phyllanthus acidus

INTRODUCTION:
Phyllanthus acidus L. Skeels is a small tropical tree that
grows up to 12 m tall, which bears pale-yellow waxy
fruits in abundance on the branches, which are edible
and tastes sour belonging to Phyllanthaceae family.[1]
Phyllanthus acidus plant is rich source of
phytochemicals with interesting biological activities
such as antioxidant[2], hepatoprotective [3], anti-
inflammatory and antinociceptive[4] activities. Volatile
constituents from the ripe fruit were analyzed by gas
chromatography-mass spectrometry. Seventy-seven
volatiles were identified: 45 terpenes, 18 esters, seven
acids, four aldehydes, two phenols, and one alcohol, of
which 66 were found for the first time in grosella.[5]
Received on 15.03.2016 Modified on 04.04.2016
Accepted on 28.04.2016 © RJPT All right reserved
Research J. Pharm. and Tech. 2016; 9(5): 559-561.
DOI: 10.5958/0974-360X.2016.00106.2
559
The scientific data regarding GC-MS analysis of
methanolic extract of Phyllanthus acidus fresh leaves
has few reported till date during literature review.
Therefore, in our present study, it was undertaken into
consideration to characterize GC-MS chromatogram for
evaluation of bioactive phytochemicals from methanolic
extract of the plant.
MATERIAL AND METHODS:
Collection of plant material:
Plant material was collected from the local areas of
Karad, Satara District, Maharashtra, India.
Preparation of extract:
Fresh leaves of Phyllanthus acidus was collected,
washed under running tap water and dried under shade
and 2 gm of dried powder of leaves of Phyllanthus
acidus was placed into a conical flask. 50ml of methanol
 Research J. Pharm. and Tech. 9(5): May 2016

was added to the conical flask and was macerated for 3
days in order to maximize the extraction. After 3 days it
was filtered through Whatmann filter paper, prepared as
methanolic stock solution and stored in a suitable
container. 1.5ml of methanolic extract was pipetted into
Eppendorf tube and sent to IICPT, Thanjavur for GC-
MS analysis.
2 min, and the total GC/MS running time was 36 min.
The relative percentage amount of each component was
calculated by comparing its average peak area to the
total areas. The mass-detector used in this analysis was
Turbo-Mass Gold-Perkin-Elmer, and the software
adopted to handle mass spectra and chromatograms was
a Turbo-Mass ver-5.2.

GC-MS analysis: Identification of phytochemical constituents:

GC-MS analysis of methanolic extract of Phyllanthus
acidus was performed using a Perkin–Elmer GC Clarus
500 system comprising an AOC-20i auto-sampler and a
Gas Chromatograph interfaced to a Mass Spectrometer
(GC-MS) equipped with a Elite-5MS (5% diphenyl/95%
dimethyl poly siloxane) fused a capillary column (30 ×
0.25 μm ID × 0.25 μm df). For GC -MS detection, an
electron ionization system was operated in electron
impact mode with ionization energy of 70 eV. Helium
gas (99.999%) was used as a carrier gas at a constant
flow rate of 1 ml/min, and an injection volume of 2 μl
was employed (a split ratio of 10:1). The injector
temperature was maintained at 250 °C, the ion-source
temperature was 200 °C, the oven temperature was
programmed from 110 °C (isothermal for 2 min), with
an increase of 10 °C/min to 200°C, then 5 °C/min to
280°C, ending with a 9 min isothermal at 280 °C. Mass
spectra were taken at 70 eV; a scan interval of 0.5 s and
fragments from 45 to 450 Da. The solvent delay was 0 to
GC-MS mass-spectrum was interpreted by using the
database of National Institute Standard and Technology
(NIST). Mass spectrum of the unknown components was
compared with the spectrum of known components
stored in the NIST library. Three major peaks of GC-MS
spectrum were ascertained while two phytochemical
components were remained unidentified.
RESULTS AND DISCUSSION:
GC-MS chromatogram analysis of the methanolic extract
of Phyllanthus acidus [Fig. 1] showed eleven peaks.
In accordance to reference mass spectra of the
constituents available in the NIST library, eleven peaks
of GC-MS spectrum revealed that squalene and others
were identified as sugar, and fatty acids [Table 1].

Figure 1: GC-MS chromatogram analysis of the methanolic extract of Phyllanthus acidus

Table 1: Phytoconstituents of methanolic extract of Phyllanthus acidus

Sr. No RT Name of Compound MF MW Peak Area %
1 3.87 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl C6H8O4 144 23.65
2 6.17 -D-Glucopyranose, 4-o--D-galactopyranosyl C12H22O11 342 31.91
3 8.07 2-Decanynoic acid C10H16O2 168 20.58
4 11.25 -D-Glucopyranoside, O--D-galactopyranosyl-(1.fwdarw.3)--D- C18H32O16 504 2.23
fructofuranosyl
5 12.13 n-Hexadecanoic acid C16H32O2 256 10.09
6 13.24 9,12-Octadecadienoic acid, methyl ester, (E,E)- C19H34O2 294 0.37
7 14.30 9,12,15-Octadecatrienoic acid, methyl ester, (Z,Z,Z)- C19H32O2 292 7.91
8 16.19 Lactose C12H22O11 342 2.37
9 18.82 1,2-Benzenedicarboxylic acid, mono(2-ethylhexyl)ester C16H22O4 278 0.21
10 22.16 Squalene C30H50 410 0.47
11 27.92 Z,Z,Z-4,6,9 Nonadecatriene C19H34 262 0.22

560
 Research J. Pharm. and Tech. 9(5): May 2016

Eleven peaks have revealed in the GC-MS spectrum of

the methanolic extract of Phyllanthus acidus. From the
results, squalene is present as major bioactive
compounds in methanolic extract of Phyllanthus acidus
fresh leaves. Other phytocomponents are found to be
fatty acids, lipid, sugar derivatives. Although it is need to
fractionate extract by using different solvents which may
helpful to explicate isolated phyto-compound and its
pharmacological assays. In our previous studies, we have
observed that squalene, 9, 12, 15-Octadecatrienoic acid
and Hexadecanoic acid are commonly found in the most
plants. [6], [7]

CONCLUSION:
The present data provides to find the molecular formula
and weight of 11 biomolecules. Further investigation
should be done to isolate bioactive compounds and their
structural elucidation and screening of pharmacological
activity in the drug development.

ACKNOWLEDGEMENT:
Authors are gratefully acknowledged to Dr. S.
Kumaravel, Quality Manager, Food Testing Laboratory
and the Director, Indian Institute of Crop Processing
Technology (IICPT), Thanjavur for providing GC-MS
facility to carry out the work.

CONFLICT OF INTEREST:
None declared.

REFERENCES:
1. Roigy Mesa JT. Diccionario Botánico de Nombres Vulgares
Cubanos. La Habana, Editorial Científico-Técnica, 1988.
2. Shilali K, Ramachandra YL, Rajesh KP, Kumara Swamy BE.
Assessing the antioxidant potential of Phyllanthus acidus bark
extracts. Int J Pharm Pharm Sci. 6(6); 2014:522-531.
3. Lee C, Peng Y, Cheng, WH, Cheng HY, Lai FNMT and Chiu TH.
Hepatoprotective effect of Phyllanthus in taiwan on acute liver
damage induced by carbon tetrachloride. Am J Chin Med. 30(3);
2006:471-482.
4. Chakraborty R, De B, Devanna N, Saikat Sen. Antiinflammatory,
antinociceptive and antioxidant activities of Phyllanthus acidus L.
extracts. Asian Pacific J Trop Biomed. 2(2); 2012: S953-S961.
5. Pino JA, Cuevas-Glory LF, Marbot R and Fuentes V. Volatile
compounds of grosella (Phyllanthus acidus [L.] Skeels) fruit.
Revista CENIC Ciencias Químicas. 39(1); 2008:3-5.
6. Durgawale PP, Hendre AS, Phatak RS. GC/MS Characterization,
Antioxidant and Free Radical Scavenging Capacities of
Methanolic Extract of Oxalis corniculata Linn: An Ayurvedic
Herb. Rasayan J Chem. 8(3); 2015: 271-278.
7. Hendre AS, Phatak RS, Durgawale PP. Phytochemical
composition of methanolic extract of pomegranate flower petals
(Punica granatum) by GC-MS analysis. Int J Pharm Sci Rev Res.
35(1); 2015: 201-202.

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