INVESTIGATION OF PHYTOCONSTITUENTS IN STEM BARK OF FICUS BENGHALENSIS LINN. USING GC-MS TECHNIQUE

INVESTIGATION OF PHYTOCONSTITUENTS IN STEM BARK OF FICUS BENGHALENSIS LINN. USING GC-MS TECHNIQUE

Shiva, Sumitra Singh ^* and Shailendra Kumar Singh

Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science and Technology, Hisar - 125001, Haryana, India.

ABSTRACT: Objective of the present study is to investigate the chemical composition of methanol extract of stem bark of Ficus benghalensis Linn. (Indian Banyan Tree). Shade and air dried stem bark were powdered and extract was prepared according to selective sequential extraction using different solvents of increasing polarity. Methanol extract of stem bark of this plant was further subjected to gas chromatography and mass spectrometry as to the best of authors’ s knowledge no published literature exist about the characterization of chemical constituents of methanol extract by GC-MS technique. Total 63 compounds were reported by GCMS studies out of which two were never reported earliar. Qualitative and quantitative presence of different biologically important phytoconstituents were reported and two new phytoconstituents were investigated. Results of this study suggested the phytopharmacological importance of reported constituents of stem bark extract of Ficus benghalensis Linn. and justify the use of Ficus benghalensis Linn. in ancient literature of Ayurveda.

Phytoconstituents, Stem bark, Ficus benghalensis Linn., GC-MS

INTRODUCTION: Ficus benghalensis Linn. ‘The national tree of India’, is native to India and commonly known as Banyan tree ¹. Its stem bark is greyish, hard, with uneven surface, on rubbing white papery flakes come out from the outer surface and inner surface is light brown without any characteristics odour ^{2 - 3}. Traditionally, stem bark is used as antioxidant ⁴, anti-inflammatory ⁵ analegesic ⁶, antipyretic ⁶, antiasthmatic ⁷ and in wound healing ⁸. According to Ayurveda the bark of Ficus benghalensis Linn. is astringent and is useful in leucorrhoea, lumbago, sores and bruises ⁹.

The bark is considered useful in burning sensation, ulcer pain, and toothache ^{10 - 11} methanol extract of Ficus benghalensis Linn. bark possess phyto-constituents with antidiabetic ¹² antiproliferative ¹³ anti-inflammatory and analgesic properties in animal models ¹⁴. Methanol extract of stem bark of Ficus benghalensis Linn. is reported to produce marked inhibitory effect on edema due to arthritis based  on various  animal models ¹⁵. The majority of activities are reported in methanol extract, thus in the present study, methanol extract of this plant was subjected to gas chromatography and mass spectrometry as to the best of author’s knowledge no published literature exists about the charac-terization of chemical constituents of methanol extract by GC-MS technique.

MATERIAL AND METHODS:

Plant Sample: The plant material (stem bark) was collected from the campus of Guru Jambheshwar University of Science and Technology, Hisar in the month of February, 2016 and was Identified by Dr. Anjula Pandey, Principal Scientist, ICAR-NBPGR, New Delhi, vide reference no. NHCP/ NBPGR /2016-1, as Ficus benghalensis Linn.

Extraction: 100 g of shade and air dried stem bark were powdered and different extracts were prepared according to selective sequential extraction using solvents of increasing polarity in order of petroleum ether, chloroform, methanol and distilled water in soxhlet extractor. Percentage yield of methanol extract was calculated and was stored in dessicator for further use.

GC-MS Analysis: GC-MS analysis of extract for identification of phytoconstituents was done using Shimadzu GCMS-QP 2010 Plus Model from Shimadzu corporation, Kyoto, Japan with Restek Rxi ^R-5Sil MS crossbond ^R similar to 5 % diphenyl /95 % dimethyl polysiloxane with 30 meter in length and 0.25 mm diameter and 0.25 micrometer in thickness.

Identification of Phytoconstituents: Bioactive phytoconstituents extracted from methanol extracts of Ficus benghalensis Linn. were identified based on GC retention time on column and by matching of this with computer software data of standards from National Institute of Standards and Technology (NIST) and Wiley Library. Molecular weights were calculated by mass spectroscopy.

RESULTS:

Percentage Yield of Extracts: Percentage yield of methanol extract of Ficus benghalensis Linn. stem bark  was 4.07 % w/w.

Physical Properties of Extracts: Methanol extract of Ficus benghalensis Linn. was brown in colour and after fluorescent study of extracts, colour changes to dark brown when viewed under short UV(254 nm) and black when viewed under long UV(365 nm) .

Phytoconstituents Reported in Extract after GC -MS: Bioactive compounds present in methanol extract of stem bark of Ficus benghalensis Linn. are shown in table no.1and as chromatogram in Fig. 1. Identification and characterization were based on their elution order in 5 Sil MS column. Elution time, molecular formula and the percentage of these phytoconstituents are also shown in the Table 1.

FIG. 1: GC-MS CHROMATOGRAM OF METHANOL EXTRACT OF FICUS BENGHALENSIS LINN.

Quinic acid (37%), α-Amyrin (4.49%), Androstan-17-one (5.53%) Lupeol acetate (2.67%) and Diglycerol (2.74%) were reported in majority according to chromatogram area percentage (Fig. 2-6). Total 63 compounds were present and two new compounds were also found in the chromatogram as their peak does not match (Fig. 7 and 8) with any compound in NIST and Wiley library. Some important phytosterols like fucosterol, lanosta-8, 24-dien-3-one, stigmasterol were present in traces. A large no. of fatty acids and their esters (8-Octadecanone) has also been identified. Tri-terpenoids like lupeol acetate, Terpenoids(α-Amyrin) were also been identified.

TABLE 1: BIOACTIVE COMPOUNDS OF FICUS BENGHALENSIS LINN. STEM BARK

FIG. 2: GC-MS CHROMATOGRAM OF QUINIC ACID          

FIG. 3: GC-MS CHROMATOGRAM OF DIGLYCEROL

FIG. 4: GC-MS CHROMATOGRAM OF LUPEOL ACETATE  

FIG. 5: GC-MS CHROMATOGRAM OF ALPHA-AMYRIN

FIG. 6: GC-MS CHROMATOGRAM OF ANDROST-17-ONE   

FIG. 7: GC-MS CHROMATOGRAM OF NEW COMPOUND-1, R.TIME-40.199, PERCENTAGE-14.40                             

FIG. 8: GC-MS CHROMATOGRAM OF NEW COMPOUND-2, R.TIME-40.013, PERCENTAGE-0.43

FIG. 9: QUINIC ACID IS SUGAR ACID AND STARTING MATERIAL FOR SYNTHESIS OF AROMATIC COMPOUNDS IN PLANTS

DISCUSSION: Chemically, quinic acid is sugar acid and starting material for synthesis of aromatic compounds in plants (Fig. 9). In a comparative study, quinic acid has been reported to be having greater anticancer potential than curcumin based on in silico study of compound ¹⁶. It is reported to exhibit antitumor activity without inducing normal cell death ¹⁷.  Another study report about significant increases in number of spleen lymphocyte cells by quinic acid ¹⁸. Hepatoprotective activity of quinic acid is reported by suitable animal models ¹⁹. Quinic acid is an antioxidant compound ²⁰. Chemically, quinic acid is cyclic polyol in nature suggesting that the six-carbon ring structure bound to a carboxylic group plays an important role in the action of polyphenols. Androstan-17-one produces antiobesity and tumor chemopreventive activity ²¹. α- Amyrin show anxiolytic, analgesic, anti-inflammatory and antidepressant effect ²². It is pentacyclic triterpenoid in nature ²³.

Another pentacyclic triterpenoid or phytosterol (Lupeol acetate) have a wide spectrum of biological activities ²⁴. In a study, lupeol acetate presents an anti-inflammatory activity in animal model by TNF-alpha and IL-2 specific mRNA regulation ^{25 - 26}. Diglycerol fatty acid esters were successfully investigated for their antibacterial activity ²⁷. Similarly other major phytoconstituents are also antioxidants in nature. Antioxidants are the substance responsible for treatment of various diseases like diabetes, cancer, inflammation etc ²⁸.

In a previous study, lanostadienyl glucosyl cetoleate and bengalensisteroic acid ester reported as the new phytoconstituents from the methanolic extract of the stem bark of Ficus benghalensis Linn. ^29. Another study report about the mechanism of action of flavonoids isolated from Ficus benghalensis Linn. having antiatherogenic potential ³⁰. GCMS act as valuable technique which provides information about bioactive phytoconstituents in any plant ³¹. According to report of a study conducted in 2014, various antioxidant compounds were reported from Ficus bengalensis Linn. tree using HPLC method ³² but our study focus on investigation of phyto-constituents using GC-MS technique. Results of the present study emphasize that the methanol extract of Ficus benghalensis Linn. is a good source of bioactive phytoconstituents  and can be used further in the field of therapeutics.

CONCLUSION: The existing knowledge regarding its phytoconstituents may be increased by the present phytochemical investigation as the two new phytoconstituents were reported from the GC-MS chromatogram of methanol extract of the stem bark of Ficus benghalensis Linn. This plant has been used in the traditional Indian System of Medicine since long time, presence of maximum phyto-constituents reported by present study also justifies the use of stem bark of Ficus benghalensis Linn. in various diseases.

ACKNOWLEDGEMENT: Study received financial support from DST, Government of India in form of DST-INSPIRE fellowship with registration no. IF140587.We would like to acknowledge JNU, AIRF Centre for providing the necessary instrumentation.

CONFLICT OF INTERST: Nil

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    How to cite this article:

    Shiva, Singh S and Singh SK: Investigation of phytoconstituents in stem bark of Ficus benghalensis Linn. using GC-MS technique. Int J Pharm Sci Res 2018; 9(6): 2478-83. doi: 10.13040/IJPSR.0975-8232.9(6).2478-83.

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