Evaluation of Phyto components in the Ethanolic Extract from Launaea Nudicaulis

K. Prabakaran¹*, M. Sathiyaseelan², S. Rameshkumar³, A. Shiva Kumar¹

¹Department of Chemistry, Sri Indu College of Engineering and Technology (Autonomous),

Sheriguda (V), Hyderabad - 501510, Telangana, India.

²Department of Chemistry, Swami Dayananda College of Arts and Science, Manjakkudi, 612610

(Affiliated to Bharthidasan University), Tamil Nadu, India.

³Department of Physics, Sri Indu College of Engineering and Technology (Autonomous),

Sheriguda (V), Hyderabad-501510, Telangana, India.

*Corresponding Author E-mail: kpraba1985@gmail.com

ABSTRACT:

Launaea nudicaulis is a significant medicinal plant with anti-inflammatory, anti-cancer, antidiabetic and antibacterial properties. In the present study, we have studied the phytochemical analysis of Launaea nudicaulis leaf extract by using NMR and GC-MS technique. According to the preliminary phytochemical analysis, identify the various active ingredients. For the purposes GC-MS study was suggested to look into the organic compounds of the unique drug Launaea nudicaulis. From the results, Launaea nudicaulis' ethanolic whole plant extract was found 11 different phytochemical compounds. The GC-MS analysis confirmed their presence various Phenyl methyl esters of 6, 9, and 12-octadecatrienoic acid (368) and Phytol (296) Molecular weight compounds. Squalene, an important chemical, was identified in the fraction using a GC-MS analysis, with an 80% match rate. By analyzing the extract using spectroscopy, ¹HNMR ¹³C NMR and GC-MS techniques, the compound squalene could be identified.

KEYWORDS: Launaea nudicaulis, Ethanolic extract, GC-MS, NMR Phytocomponents.

INTRODUCTION:

To address their basic healthcare needs, communities in impoverished nations, mainly rely on traditional herbal remedies. Herbal medications are becoming more and more used as supplementary and alternative therapies in many developed nations. Biological and pharmacological processes occur in plants. Numerous biologically active substances that have been widely employed as medicines, food additives, tastes, insecticides, colourants, perfumes, and chemicals are produced by plants as their secondary metabolites¹. With more people seeking treatments and health strategies, free from side effects brought on by synthetic pharmaceuticals, medicinal plants are transitioning from being utilized in a fringe capacity to being more widely accepted².

There is a need to re-evaluate the treatment efficacy of herbal medicines because they have been used as bioactive molecules for thousands of years³. Biologically active secondary metabolites can provide It is essential supply Various structural arrangements can be found and characteristics⁴. Notable Compounds that belong to this group include phenols, saponins, cyanogenic glycosides, and phenolic glycosides⁵. For additional chemical and pharmacological investigation, it has been found that in vitro screening the raw plant extracts containing An extremely useful property⁶.

Purgative drugs that are extracted from plants or created using technology may be preferred due to their strong activity against human illness, but also have problems. At parallel doses or percentages of the active ingredient, they seldom ever exhibit Similar to crude extract in terms of activity⁷. Only 6% of the estimated 6 lakh plant species on Earth have been studied, which highlights the opportunities and difficulties presented to phytochemists⁸.

Numerous Launaea species are widely recognised in traditional medicine for their wide range of therapeutic characteristics, including diuretic, aperients, soporific, galactagogue, febrifuge, and stomachic^9-11. They also contain anticancer, insecticidal, cytotoxic, and antioxidant activity. The ethanolic fraction of the Launaea nudicaulis leaf extract was chosen as the study's subject because there is no available information on the phytoconstituents of this fraction. The goal of this work is to employ Analyzing the chemical composition of a sample using GC-MS active fraction of the Launaea nudicaulis leaf extract, which may give insight into its usage in traditional medicine. The separation is performed by GC-MS after a simple solvent extraction, we discovered the active compounds found in this medicinally useful plant. At the same time, ¹H and ¹³C NMR have been widely employed for identification and evaluation, as well as quality assessment and authenticity.

MATERIALS AND METHODS:

Plant Materials:

Sengipatti, Thanjavur District, Tamil Nadu, was selected as the source of the fully mature Launaea nudicaulis leaf. The identified plant part this authentication was provided by Dr. S. Mahesh, Department of Microbiology, Marudupandiyar College, Thanjavur.

Plant sample extraction:

After soaking 20 gm powdered plant material after overnight immersion in 50 ml of absolute alcohol, For the removal of sediments and residues, Whatman filter paper No.41 and 2gm sodium sulphate are used. In order to filter the solution, 100% alcohol is sprayed on the filter paper and the sodium sulphate. 1ml of nitrogen gas is then bubbled into the filtrate to concentrate it. Many of the medicinal qualities of plants are attributed to complex chemicals or active principles. The current study's major goal was to examine the chemical makeup and Standardization of herbal medications based on phytoconstituents found in plants ¹².

Figure 1: Launaea Nudicaulis

GC –MS analysis:

The Perkin Elmer GC Clarus 500 system, which includes an AOC-20i autosampler and gas chromatograph connected to a mass spectrometer equipment, was set up for GC-MS analysis using the following settings: Helium gas (99.999%) was utilised as the carrier gas at a constant flow of 1ml /min and an injection volume of 0.5 I was used in column Elite-1 fused silica capillary column (30 x 0.25mm ID x 1Mdf, made entirely of dimethylpolysiloxane) (split ratio of 10:1) Temperatures of the ion source are 280°C., while the injector is 250°C. With 0.5 second scanning intervals and fragment sizes ranging from 40 to 450 Da, mass spectra at 70 eV were obtained. The GC takes 36 minutes to complete in its entirety. The chromatogram was produced using Turbo Mass Ver 5.2.0 and modified software for detecting mass spectra. Test materias were named, molecular weighted, and structurally composed and identified by comparing the spectrum of the unknown component to the spectrum of known components recorded in the NIST and MS libraries. The mobile phase is made up of helium gas. The observed outcomes were deciphered^13.

NMR analysis:

On a Bruker AMX 400 equipment (Bruker Company, Faelladen, Switzerland), conventional pulse sequences operating at 400 MHz for ¹H NMR and ¹³C NMR investigations have been carried out. The coupling constants (J) are reported in Hz and the chemical shifts are given in ppm around TMS as the internal reference material¹⁴.

Table: 1 The components identified in the ethanolic leaf extract of Launaea nudicaulis (GC MS study)

Sl. No	RT	Name of the compound	Molecular	MW	Peak Area %
1	11.14	3,7,11,15-Tetramethyl-2-hexadecen-1-ol	C₂₀H₄₀O	296	7.58
2	12.13	Tridecanoic acid, methyl ester	C₁₄H₂₈O₂	228	0.55
3	12.89	Decanoic acid, ethyl ester	C₁₂H₂₄O₂	200	1.73
4	14.12	2-Cyclopentene-1-undecanoic acid, (+)-	C₁₆H₂₈O₂	252	0.30
5	14.20	9,12,15-Octadecatrienoic acid, methyl ester, (Z, Z, Z)-	C₁₉H₃₂O₂	292	1.35
6	14.34	Phytol	C₂₀H₄₀O	296	15.97
7	15.05	9,12,15-Octadecatrienal	C₁₈H₃₀O	262	2.62
8	20.07	1,2-Benzenedicarboxylic acid, diisooctyl ester	C₂₄H₃₈O₄	390	13.40
9	23.81	Squalene	C₃₀H₅₀	410	47.71
10	27.96	Vitamin E	C₂₉H₅₀O₂	430	6.79
11	30.91	6,9,12-Octadecatrienoic acid, phenylmethyl ester, (Z, Z, Z)-	C₂₅H₃₆O₂	368	2.00

Table: 2 Activity of Phyto-components identified in the ethanolic leaf extracts of the Launaea nudicaulis by GC-MS

Sl. No.	Retention Time	Name of the Compound	Biological activity*
1	14.20	9,12,15-Octadecatrienoic-acid, methyl ester, (Z,Z,Z)	Anti-viral activity ¹⁵
2	14.34	Phytol	Cancer-Preventive ¹⁶
3	12.89	Decanoic acid, ethyl ester	Nematicide, pesticide and flavor,
4	23.81	Squalene	Antibacterial, Antioxidant, Antitumor, Cancer-Preventive, Chemopreventive, Immunostimulant and Lipoxygenase-Inhibitor.
5	27.96	Vitamin E	Hepatoprotective,Hypocholesterolemic, Hypoglycemic and Immunostimulant ^{17, 18}
6	30.19	Phenyl methyl ester of 6,9,12-octadecatrienoic acid	anticancer, cancer preventive and antioxidant activity

*Source: Dr.Duke's phytochemical and ethnobotanical databases [Online database]

RESULTS AND DISCUSSION:

Plants contain and produce aromatic compounds, the majority of which are phenols. These operate as defensive systems for plant extracts against insects and herbivores. Hepatotoxic, anti-fungal, anti-inflammatory, and anti-ulcer activities are biological examples of reactions. The constituents of the test materials' molecular weight and structure were determined. The biological processes mentioned in Table 2.

GC-MS ANALYSIS:

The eleven chemicals were found in Launaea nudicaulis leaf extracts using GC-MS analysis Table 1. The prevailing compounds were 3, 7, 11, 15, -Tetra methyl -2-hexdeen-1 (11.4), Tridecanoic acid, methyl ester, Decanoic acid, ethyl ester (12.89), 2-cyclopentene-1-undecanoic acid (14.12), 9,12,15-octadecatrienoic acid, methyl ester (Z,Z,Z) (14.20), Phytol (14.34), 9,12,15-octadecatrienal(15.05), 1,2 Benzenedicarboxlic acid diisooctyl ester (20.07) (26), Squalene (23.83), Vitamin E (28.03), Phenyl methyl ester of 6,9,12-octadecatrienoic acid (30.19)¹⁹. It is clear that the leaf extracts include a variety of phytocompounds that might be useful in the pharmaceutical industry due to their suitable biological properties. However, more research like bio-prospecting is necessary to support its biological features support its biological features, however, more research like bio-prospecting is necessary ²⁰.

NMR Analysis:

The presence of six carbon peaks in the olefinic carbon range (C 124.42, 124.45, 125.03, 131.18, 135.06, 135.20 ppm) indicates the presence of three double bonds in the molecule. C-17 (C 131.18ppm), an olefinic quaternary carbon from the HMBC spectrum,connected with protons on C-6, 9, and 11. The presence of singlet splitting in the protons linked to C-6, H 1.59ppm (s), and C-9, H 1.68ppm (s), suggests that C-6 and C-9 were directly bound to the olefinic carbon and that there were no surrounding protons. Additionally, C-6 had a connection with protons at H 5.11ppm, showing that C-6 was not the only olefinic carbon linked with a proton (H-C=C) at two or three different bonds. It was discovered that the proton at H 5.11ppm correlates with the proton at H 2.05ppm and protons at H 1.97ppm²¹. Squalene with a symmetrical structure has been introduced as the suggested structure and comparing this with the declared literature value will prove this.

TLC:

A pale yellow substance (33.01 mg) with Rf value of 0.32 (20% DCM: 80% hexane) was found in the compound.

CONCLUSION:

In the current study, the presence of chemical components of an ethanolic leaf extracts from Launaea nudicaulis was identified by using NMR and Gas Chromatogram-Mass Spectrometry analysis. Squalene in samples may be detected by an invention of the 13C and 1H NMR spectroscopic analysis. The presence of numerous bioactive components proved that many disorders were treated with the plant by traditional healers. Results can be obtained by isolating certain phytochemical components and submitting them through biological action. Launaea nudicaulis is thought to contain a variety of bioactive substances. It is suggested as a plant with the potential for use in pharmaceuticals. Future research will be required to completely determine Agricultural products, its bioactivity, and its toxicity profile.

AUTHOR CONTRIBUTIONS:

All authors contributed to the study conception and design, Dr. K. Prabakaran: Conceptualization, Methodology, Investigation, Writing – original draft, Writing – review and editing, Supervision.

CONFLICT OF INTEREST STATEMENT:

The author declared no conflict of interest.

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Received on 07.02.2022 Modified on 23.11.2022

Research J. Pharm. and Tech 2023; 16(11):4989-4992.

DOI: 10.52711/0974-360X.2023.00807