GC-MS INVESTIGATION OF PHYTOCOMPONENTS PRESENT IN ETHANOLIC EXTRACT OF PLANT ICHNOCARPUS FRUTESCENS (L.) W. T. AITON AERIAL PART

GC-MS INVESTIGATION OF PHYTOCOMPONENTS PRESENT IN ETHANOLIC EXTRACT OF PLANT ICHNOCARPUS FRUTESCENS (L.) W. T. AITON AERIAL PART

Ashutosh Meher ^{* 1}, Biswaranjan Behera ² and Bikash Kumar Nanda ³

The Pharmaceutical College ¹, Barpali, Bargarh - 768029, Odisha, India.

Kanak Manjari Institute of Pharmaceutical Sciences ², Rourkela - 769015, Odisha, India.

Jayapur College of Pharmacy ³, Jayapur - 764002, Odisha, India.

ABSTRACT: Ichnocarpus frutescens (Apocynaceae) generally renowned as Kali Sariva in Sanskrit, Krishna Sariva in Hindi which is a large, evergreen, red woody climber, native to India, Java, China, Southeast Asia, Ceylon, Northern Australia and found ascending to an altitude of 4,000 ft. Different tribes of India are used this plant as a substitute of Indian Sarsaparilla (Hemidesmus indicus). It has been used conventionally in various diseases and disorders of human being that is a headache, wound, fever, tongue ulcers, cramps, night blindness, stomach pain, bone fracture, skin infection, diabetes, liver disorders, as alterative, tonic, diuretic and diaphoretic. Pharmacologically plant shows different activities include anti-diabetic, anticarcinoma, antiurolithiatic, antiobesity, analgesic, antipyretic, anti-inflammatory, skeletal muscle relaxant, antitumor, hepatoprotective. The present study was aimed to identify the phytocomponents present in I. frutescens using GC-MS analysis. The aerial plant part was extracted successively by solvents according to their increasing order of polarity using Soxhlet apparatus. The phytocomponents of ethanolic extract was identified by GC-MS investigation and found 19 number of phytocomponents. The major constituents of the extracts were 3-O-Methyl-d-glucose, 3, 4, 6-Tri-O-methyl-d-glucose, 2-O-Methyl- D-mannopyranose, Myo-In-ositol, 4-C-methyl and Myo-In-ositol, 2-C-methyl. This study is the prime characterization of the phytocomponents of plant data that indicates the ethanolic extract has remarkable activities.

Ichnocarpus frutescens, Apocynaceae, Phytocomponents, GC-MS

INTRODUCTION: In various traditional systems of medicine, plants are used as a remedy for different ailments. In general, plants are utilized by Vaidya, tribals and local healer as medicine and are improved by time to time for better efficacy. Different species of plant restrain a huge number of phytocomponents which have medicinal values but are a virgin till date.

Ichnocarpus frutescens (Linn.) W. T. Aiton (Apocynaceae) is well-known as Krishna Sariva in Ayurvedic system of medicine ¹. It is a large, evergreen, laticiferous, woody creeper with rusty red appearance, found almost all over India, ascending to an altitude of 4,000 ft ² and also found in Ceylon, China, Java and Australia ³. The plant root is used as an alternative of Indian sarsaparilla (Hemidesmus indicus).

Conventionally the plant has been used in headache, wound, fever ², tongue ulcers, cramps, night blindness, headache ⁴, bone fracture, skin infection, diabetes and liver disorders ⁵. Pharmacologically plant showed different activities include antidiabetic, anticarcinoma, antiurolithiatic, anti-obesity, analgesic, antipyretic, anti-inflammatory, skeletal muscle relaxant, antitumor, hepatoprotective ^{6, 7, 8} anticonvulsant activity ⁹. The plant may show different activity due to the presence of phytocomponents phenylpropanoids, phenolic acids, coumarins, flavonoids, sterols and pentacyclic triterpenoids, i.e. Δ¹²-dehydrolupanyl-3β-palmitate, lupeol acetate, friedelin, friedelinol, Δ¹²-dehydrolupeol, oleanolic acid, nonane, 5-hydroxyoctacosan-25-one, dotriacontanoic acid, sitosterol and sitosterol palmitate ¹⁰. With this background, the present study was aimed to identify the phytocomponents from the ethanolic extract of I. frutescens aerial part by using GC-MS investigation.

MATERIAL AND METHODS:

Collection and Preparation of Plant Materials: Fresh aerial parts of the plant were collected from an adjoining area of Barpali, Odisha, India and identified by Botanical Survey of India, Kolkata, India, bearing Ref. no: CNH/I-I(5)/2009/Tech. II/35. The aerial parts of the plant are dried under shade and pounded into powder using a mechanical grinder. The powdered material was stored in an airtight container until use.

Preparation of Extract: The powdered material was extracted with petroleum ether (60-80 °C), chloroform, ethyl acetate and ethanol successively in the Soxhlet apparatus. The extracts thus obtained were concentrated in a rotary evaporator and stored in the refrigerator at 4 °C for further use. The extracts were employed for GC-MS analysis.

GC-MS Investigation: The investigation of I. frutescens ethanolic extract was performed by using GC-MS instrument Thermo Trace 1300GC coupled with Thermo TSQ 800 Triple Quadrupole MS. The investigational circumstance of GC-MS system was as the following condition; TG 5MS (30 m × 0.25 mm ID × 0.25 µm) column composed of 5% phenyl methylpolysiloxane, helium gas was used as carrier gas at a constant flow rate of 1.0 ml/min and an injection volume of 1.0 µl was employed (split ratio of 10:1, injector temperature operated at 250 °C; ion-source temperature 280 °C and the oven temperature was programmed from 60 °C (isothermal for  2 min.) with an increase of 10°C/ min. to 280 °C (isothermal for 10 min.). Mass spectroscopy (MS TSQ 8000) was taken at 70 eV; a scanning interval of 0.5 sec and fragments from 40 to 550 Da.

Identification of Compounds: The phyto-components were identified by interpretation of mass-spectrum with the library data of National Institute standard and Technology (NIST). The name, molecular formula and molecular weight of the components were established.

RESULTS: The GC - MS chromatogram of the I. frutescens ethanolic extract showed the presence of nineteen compounds. The identified phyto-components name, retention time (RT), peak area (%), molecular formula, molecular weight, nature of compound and activities are presented in Table 1.

TABLE 1: PHYTOCOMPONENTS IDENTIFIED IN THE ETHANOLIC EXTRACT OF I. FRUTESCENS AERIAL PART BY GC-MS

The phytocompound prediction is based on Dr. Duke’s Phytochemical and Ethnobotanical Databases by Dr. Jim Duke of the Agricultural Research Service, USDA. The results showed the presence of dodecane, 2,6,11-trimethyl, undecane, tetradecane, pentadecane, 3-O-Methyl-d-glucose, 3, 4, 6- Tri-O-methyl-d-glucose, 2- O-Methyl- D-mannopyranosa, Myo-In-ositol, 2-C-methyl (Iso-mytilit), Myo-In-ositol, 4-C-methyl, hexadecanoic acid, ethyl ester (Ethyl palmitate), pentadecanoic acid, ethyl ester, hexadecanoic acid, 2-methyl, methyl ester, phytol, 3, 7, 11, 15-Tetramethyl-2-hexadecen-1-ol, 1-Hexadecen-3-ol, 3, 5, 11, 15-tetramethyl (Isophytol), 13-Docosenamide,(Z),  9-Octadecenamide, (Z), 8-Methyl-6-nonenamide. The spectrum profile of GC-MS confirmed the presence of 19 components and the chromatogram of the phytocomponents are illustrated in Fig. 1-7.

FIG. 1: GC-MS CHROMATOGRAM OF Dodecane, Dodecane, 2, 6, 11-trimethyl and Undecane (RT 7.85)

FIG. 2: GC-MS CHROMATOGRAM OF Tetradecane, Pentadecane (RT 9.77)

FIG. 3: GC-MS CHROMATOGRAM OF 3-O-Methyl-d-glucose, 3, 4, 6- Tri-O-methyl-d-glucose, 2- O-Methyl- D-mannopyranosa (RT 12.26)

FIG. 4: GC-MS CHROMATOGRAM OF Myo-In-ositol, 2-C-methyl (Iso-mytilit), Myo-In-ositol, 4-C-methyl (RT 12.64)

FIG. 5: GC-MS CHROMATOGRAM OF hexadecanoic acid, ethyl ester (Ethyl palmitate), pentadecanoic acid, ethyl ester, hexadecanoic acid, 2-methyl, methyl ester (RT 14.31)

FIG. 6: GC-MS CHROMATOGRAM OF phytol, 3, 7, 11, 15-Tetramethyl-2-hexadecen-1-ol, 1-Hexadecen-3-ol, 3, 5, 11, 15-tetramethyl (RT 15.10)

FIG. 7: GC-MS CHROMATOGRAM OF 13-Docosenamide, (Z), 9-Octadecenamide, (Z), 8-Methyl-6-nonenamide (RT 19.66)

DISCUSSION: In the present study, the GC-MS investigation of the ethanolic extract of Ichnocarpus frutescens showed the presence of nineteen compounds. In terms of percentage quantity, 3-O-Methyl-d-glucose, 3, 4, 6-Tri-O-methyl-d-glucose & 2-O-Methyl-D-mannopyranosa were found to be a prime percentage in the extract. The major phytocompound 3-O-Methyl-d-glucose has preservative activity. Antimicrobial, anti-inflammatory, anticancer and antidiabetic activities are shown by tetradecane, phytol and 3, 7, 11, 15-Tetramethyl-2-hexadecen-1-ol similarly 8-Methyl-6-nonenamide showed antimicrobial and anti-inflammatory activities. The other identified phytocomponents are responsible for various activities except 3, 4, 6-Tri-O-methyl-d-glucose, 2-O-Methyl- D-mannopyranosa, Myo-In-osito- l, 2-C-methyl (Isomytilit), Myo-In-osito-l, 4-C-methyl, pentadecanoic acid, ethyl ester and hexadecanoic acid, 2-methyl, methyl ester.

CONCLUSION: The present investigation concluded that the ethanolic extract has a number of bio-active phytocomponents responsible for many biological activities and justify the use of the plant for different diseases and disorders of a human being by Vaidya, tribals and local healer or traditional practitioners as medicine. So, further separation, isolation and characterization of individual phyto-components from the plant, may be undertaken to discover novel drugs and their therapeutic actions to treat various ailments.

ACKNOWLEDGEMENT: The authors are thankful to Head, CIL Punjab (India) for providing spectral and analytical data.

CONFLICT OF INTEREST: We declare that we have no conflict of interest.

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    How to cite this article:

    Meher A, Behera B and Nanda BK: GC-MS investigation of phytocomponents present in ethanolic extract of plant Ichnocarpus frutescens (L.) W. T. Aiton aerial part. Int J Pharm Sci & Res 2019; 10(10): 4711-16. doi: 10.13040/IJPSR.0975-8232.10(10).4711-16.

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