GC-MS Analysis of the Methanolic extract of Cuscuta reflexa Roxb. and Gymnema sylvestre (Retz.) R. Br. ex. Sm.

 

Bishnupada Biswal¹*, Susanta Kumar Panda²

¹Department of Pharmacy, Biju Patnaik University of Technology, Rourkela, Odisha, India.

²Department of Pharmacy, Royal College of Pharmacy and Health Sciences, Berhampur, Odisha, India.

 

ABSTRACT:

The present study was designed to determine the bio-active compounds in the aerial parts of the methanolic extract of Cuscuta reflexa Roxb. and Gymnema sylvestre (Retz.) R. Br. ex. Sm. GC-MS analysis of the extracts was performed on a GC–MS equipment (Thermo Scientific Co. Thermo GC-TRACE ultra ver.: 5.0, Thermo MS DSQ II). The phytochemical tests of extracts of both the plants confirmed the presence of alkaloids, glycosides, saponins, steroids, terpenoids, phenols, tannins and flavonoids. The GC-MS analysis has shown the presence of eleven and thirteen different phytochemical compounds in the methanolic extract of Cuscuta reflexa and Gymnema sylvestre, respectively. From the results, it is evident that both the plant contains various phytocomponents and both the plants are recommended as medicinal plant for the treatment of some diseases.

 

 

 

INTRODUCTION: 

Plants as a source of medicine which can be produce new drugs for the benefit to mankind. India is the largest producer of medicinal herbs and is appropriately called the botanical garden of the world¹. The use of medicinal plant products as therapeutic agents and is mainly involved or applied in the traditional type of medicine and especially seen in Ayurvedic and Unani treatment^{2, 3}. Today, nearly 88% of the global populations switch to plant derived medicines as their first line of defense for maintaining health and combating diseases⁴. In last few decades, there is an overwhelming demand of natural medicine around the world, which has promoted the well organized and systemic research and development for the search of newer pharmaceutical drugs from medicinal plants⁵. Herbal medicines are accepted to be more secure than synthetic medicine⁶.

 

In the last few years, gas chromatography mass spectrometry (GC-MS) has become firmly established as a key technological platform for secondary metabolite profiling in both plant and non-plant species^7-9.

 

Therefore, in GC-MS analytical research, traditional medicine has been inherited interest in identify for important biologically active compounds for the treatment of various diseases. The traditional medicine involves the use of different plant extracts or the bioactive constituents¹⁰.Chemical constituents embedded in the herbs such as carbohydrates, flavonoids, alkaloids, glycosides and lignans have been found to possess medicinal benefits¹¹. Scientific evaluation of plants has often shown that active principles in these are responsible for therapeutic success¹².

 

Cuscuta reflexa Roxb. (Family: Cuscutaceae) is known as “Akashabela, Amarbela kasur” in Hindi, also used in India¹³. This plant is tropical and sub-tropical herb which is depending on the host plants for originated as parasite weed. Antibacterial properties have been observed in stem of Cuscuta reflexa Roxb. (C. reflexa) which is used internally to treat fever and externally in itching¹⁴. This plant also exhibits anticancer and anti-inflammatory activities¹⁵, diuretic activity¹⁶ and anti-HIV activity¹⁷.

 

Gymnema sylvestre (Retz.) R. Br. ex. Sm. (G. sylvestre) is an indigenous herb, belonging to the class dicotyledonous of the family Asclepiadaceae¹⁸. The plant is a good source of a large number of bio-active substances¹⁹. It has deep roots in history, being one of the major botanicals used in Ayurvedic system of medicine to treat conditions ranging from diabetes, malaria to snakebites²⁰. According to the Ethanomedical information of Gymnema sylvestre, it is being used as anti-diabetic, stomachic, stimulant, laxative and diuretic²¹.

 

Plants are important sources of potentially useful constituents for the development of new therapeutic agents because most of them are safe with little side effects²². The GC-MS analysis of the methanolic extract of C. reflexa and G. sylvestre are help to investigate the various compounds available in these plants which was not reported still today. Hence, the aim of the present study is to identify the chemical compounds of the methanolic extracts of C. reflexa and G. sylvestre by GC-MS.

 

MATERIAL AND METHODS:

Collection and authentication of plant materials:

The aerial parts of C. reflexa (Convolvulaceae) and G. sylvestre (Apocynaceae) were collected in the month of December 2020 and March 2021, respectively from various areas of Purba Medinipur District fields of West Bengal and authenticated by Botanist K. Karthigeyan and Vinay Ranjan (Scientist-E, Botanical Survey of India, Kolkata, West Bengal, India), respectively. Plant’s herbarium was prepared and preserved in the Central National Herbarium, Botanical Survey of India, Kolkata, West Bengal, India.

 

Preparation of extracts:

110 g of dry aerial plant parts, suitably powdered, of each drug was extracted with 700 ml. of methanol by Soxhlation process ²³. The extracts obtained were concentrated under reduced pressure and air dried²⁴. The concentrated extracts were further dried in desiccators²⁵ which contain fused calcium chloride. After that, the extracts were stored in a refrigerator at 4°C for further use. 

 

Identification of phyto-constituents by preliminary phytochemical tests:

The methanolic extracts were tested for alkaloids, glycosides, saponins, steroids, terpenoids, phenols, tannins, flavonoids, proteins and amino acids and Pholbatannin^26-30.

 

GC-MS study:

GC-MS analysis of methanol extracts were performed on a GC-MS instrument (Thermo Scientific Co. Thermo GC-TRACE ultra ver.: 5.0, Thermo MS DSQ II). Experimental conditions of GC-MS system were as follows: DB 35-MS capillary standard non-polar column, dimension: 30Mts, ID: 0.25 mm, Film thickness: 0.25µm. Flow rate of mobile phase (carrier gas: He) was set at 1.0 ml/min. In the gas chromatography part, temperature programme (oven temperature) was 50°C raised to 260°C at 10°C/min and injection volume was 1 µl. Samples were run fully at a range of 50–650 m/z and the results were compared by using Wiley Spectral library search programme.

 

RESULTS AND DISCUSSION:

Alkaloids, glycosides, saponins, steroids, terpenoids, phenols, tannins and flavonoids phytoconstitutents were present in the methanolic extracts of C. reflexa and G. sylvestre.

 

The Phytochemical investigation of the methanolic extract of C. reflexa showed 11 various compounds by GC-MS analysis and it’s confirmed through GC-MS spectrum with different retention times (RT) as illustrated in Figure 1 and identified compounds were shown in Table-1. Benzoic acid, methyl ester, 2-Propenoic acid, 1,7,7-trimethylbicyclo[2.2.1]hept-2-yl ester, exo-, Irgacure 184, 6-Octadecenoic acid, methyl ester, Cholestano[7,8-a]cyclobutane, 3-methoxy-6-oxo-2'-methylene, Thymolphthalein, Cholest-2-eno[3,2-a]naphthalene, Diethyl 2-(2-furyl)-4-hydroxy-4-methyl-6-oxo-1,3-cyclohexaned icarboxylate tbdms, 2,5-Dichloro-3-ethoxycarbonyl-4,6-di(4-methylphenylth io)pyridine, EPINEPHRINE-TETRATMS, Pregnane-3,11,20-trione, (5á) were present in the methanolic extracts of C. reflexa.

Table 1. Compounds identified in the extract of Cuscuta reflexa Roxb. by GC-MS

RT: Retention time (min).

 

Figure 1. GC-MS Chromatogram of methanolic extract of Cuscuta reflexa Roxb.

 

 

Figure 2. GC-MS Chromatogram of methanolic extract of Gymnema sylvestre (Retz.) R. Br. ex. Sm

 

Table 2. Compounds in the extract of Gymnema sylvestre (Retz.) R. Br. ex. Sm. by GC-MS

RT: Retention time (min).

 

The GC-MS analysis of methanolic extract of G. sylvestre detected 13 various compounds which were confirmed by GC-MS spectrum with various retention times as illustrated in Figure 2 and identified compounds were shown in Table-2. Hexasiloxane, 1, 1, 3, 3, 5, 5, 7, 7, 9, 9, 11, 11-dodecamethyl-, Naphthalene, 1, 2, 3, 4, 4a, 5, 6, 8a-octahydro-7-methyl-4-methylene-1-(1 -methylethyl)-, (1à,4aá,8aà)-, 1,6,10-Dodecatrien-3-ol, 3,7,11-trimethyl-, (E)-, Globulol, Methanone, (1-hydroxycyclohexyl)phenyl-, Benzo[c][2,7]naphthyridinone, 2-Hexadecen-1-ol, 3, 7, 11, 15-tetramethyl-, [R-[R*,R*-(E)]]-, Docosane, Nonacosane, 4à,14à,24-trimethyl-9(11)-cholesten-3á-yl acetate, 6-Chloro-2-(2-hydroxyethylamino)-4,8-bis(4-methoxya nilino)pyrimido[5,4-d]pyrimidine,   Methyl [4-(5',6',7',8'-Tetrahydro-5',5',8',8'-tetramethyl-2'-anthra cenyl)phenyl] sulfone, 1,2-Benzenedicarboxylic acid, dioctyl ester were shown in the methanolic extract of G. sylvestre.

 

CONCLUSION:

The aerial parts of plant extracts contained significant levels of phytoconstitutents which were identified by GC-MS as per present investigation. The GC-MS analysis result has shown in the C. reflexa and G. sylvestre methanolic extracts which contains eleven and thirteen of different bio-active compounds, respectively.

 

Thus, isolation of individual phytocomponents may be useful for biological activity and can be established as a new drug for the treatment for any disease because these compounds can be responsible for different therapeutic and pharmacological properties. Further research to determine its bioactivity, toxicity profile, pre-clinical tests and clinical studies are necessary for drug discovery and development. Screening of pharmacological study is under progress.

 

CONFLICT OF INTEREST:

The authors declare that there is no conflict of interests regarding the publication of this article.

 

ACKNOWLEDGEMENTS:

The authors express their gratitude to the Management, Bharat Technology, Uluberia, Howrah, West Bengal for providing their continuous support throughout the work. The authors are also grateful to Prof. (Dr.) Lakshmi Kanta Kanthal and Mrs. Anima Jena for their continuous encouragement and valuable inputs and co-operation while carrying out this study.

 

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Accepted on 07.02.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2023; 16(1):18-22.