GC-MS Analysis of Phytochemicals and Green synthesis of Silver nanoparticles from Amaranthus parganensis

Smarika Chauhan, Swamynathan G*

Department of Biotechnology, College of Science and Humanities, SRM Institute of Science and Technology, SRM Nagar, Kattankulathur, 603203, Kanchipuram, Chennai, TN, India.

*Corresponding Author E-mail: smarikachauhan25@gmail.com, swamynathanganesan.in@gmail.com

ABSTRACT:

There are different kinds of flora found across the world. Diverse variety of plants are there like ornamental plants, indoor plants, vegetable crops, medicinal plants, wild plants etc. Every plant has its own significance and utilizations in diverse fields. Mainly medicinal plants have therapeutic uses in the drug development process. But even other type of plants such as vegetable crops have been found to have medicinal values. Therefore, green vegetables are essential part of our diet from ancient times. This study focuses on evaluating the properties of a leafy vegetable herb namely Amaranthus parganensis. The biologically active compounds that were found through GC-MS revealed the presence of phytol; squalene; 9-Octadecenoic acid (Z)-, 2-hydroxy-1-(hydroxymethyl) ethyl ester; N-Hexadecanoic Acid; 3,4,5-Trimethoxy-2',4'-diaminodiphenylsulfide; 3,7,11,15-Tetramethyl-2-hexadecen-1-ol and Ethoxy(methoxy)methyl silane in varied amounts. Furthermore, silver nanoparticles which has therapeutic applications may be synthesized from Amaranthus parganensis. We conclude through this study that Amaranthus parganensis has bioactive compounds which may have therapeutic value. Also, we have shown that the plant could be a potential source for silver nanoparticles synthesis which may have therapeutic applications.

KEYWORDS: Amaranthus parganensis, Phytochemicals, GC-MS, Silver nanoparticles, Green synthesis.

INTRODUCTION:

Across the world, plants with therapeutic qualities have been utilised broadly to avert numerous diseases in traditional herbal medicine systems^1-3. Due to wide availability of plants with medicinal properties, the demand of these plants is on the rise for the production of cost-effective drugs⁴. Natural holistic medicines that are based on medicinal plants are still used by the local people in many countries for therapeutic purposes. Many countries all over the world follow and have their own indigenous common remedies for treatment of diseases⁵. Several properties such as antioxidant, antibacterial and therapeutic properties of natural plants have been verified which forms the basis for its use in medicines.

After examination of these properties, medicinal plants are used in pharmacology and therapy to treat several diseases^6,7. Different biological compounds with a variety of activities are present in natural plants which can be exploited^8,9. Many types of diseases are treated by active compounds present in parts of plants like roots, stem, leaves and bark¹⁰.

Natural compounds of plants have been utilized in conventional system of medication to cure different infections and disorders. There are many plants whose total medicinal value are even now unknown and Amaranthus spp. is one among them¹¹. Amaranthus parganensis occurs widely in different regions of West Bengal and is cultivated in open areas as a vegetable crop¹². Many researchers have focused on identifying the various compounds present in the Amaranthus plant as each part of this plant may have therapeutic uses. The current focus is to investigate the antioxidant as well as anti-inflammatory properties of this plant so that it can be used as a functional food with medicinal value. Phytochemical study of different Amaranthus spp. has found the existence of various biologically active components such as polyphenols, flavonoids, steroids, saponins, carbohydrates, tannins, terpenoids and fats^13-16. Majority of the Amaranthus spp. have antimicrobial, anti-malarial, antioxidant, anti-inflammatory, anti-cancer and anti-diabetic activity in nature. Protection from many critical disorders and infections can be achieved through the activity of polyphenols that are present in Amaranthus¹¹. Therefore, it is an essential plant that could be used for the development of plant-based drugs. In this study, we have focused on identifying the phytochemicals present in Amaranthus parganensis which could have medicinal properties. Also, we have carried out the green synthesis of silver nanoparticle from Amaranthus parganensis which could be used for the development of therapeutic drugs.

MATERIALS AND METHODS:

Collection and extraction of plant sample:

Amaranthus parganensis was collected from West Bengal, India. The collected leaves were carefully washed with water to remove adherent dust, dried in shade, coarsely powdered and stored in air tight zip lock cover bags^17,18. 10g of powdered leaves of Amaranthus parganensis was extracted successively with 100ml of 99% ethanolic solution. The mixture was then incubated at 4℃ for 3 days and was then filtered and air dried in petri dishes. After complete drying the solid extract was scrapped to powder form and stored for further analysis ¹⁹. Another extraction method used was 10g of powdered leaves of Amaranthus parganensis was homogenized with 100ml of 99% ethanolic solution and contents was kept for incubation at 4℃ for 2 days. This suspension was then filtered. The filtered part was taken and solvent was evaporated in a rotator evaporator at 40℃. The extract was used for further analysis¹⁹.

GC-MS Profiling of phytocompounds:

Amaranthus parganensis was analyzed for various phytochemicals using the ethanolic extract by performing the GC-MS analysis^20-22.

Synthesis and Characterization of silver nanoparticles:

The silver nanoparticles were synthesized via preparing the ethanolic plant extract with 100mg/ml concentration and 1mM AgNO₃ solution. 10ml of 1mM of silver nitrate was added to 2ml of ethanolic leaf extract. The blend was incubated at 27℃ for 2 hrs. Colour change to reddish-brown was observed in the chemical mixture ^23,24. This indicates the synthesis of silver nanoparticles. The synthesized silver nanoparticles were centrifuged and dried for characterization²⁵. The synthesized nanoparticles of Amaranthus parganensis were characterized utilizing SEM and EDX ^26-29.

RESULTS:

Gas chromatography-mass spectroscopy profile:

In Amaranthus parganensis, the biologically active compounds determined through GC-MS includes Ethoxy(methoxy)methyl silane; 3,7,11,15-Tetramethyl-2-hexadecen-1-ol; 3,4,5-Trimethoxy-2',4'-diaminodiphenylsulfide; N-Hexadecanoic Acid; Squalene; Phytol and 9-Octadecenoic acid (Z)-, 2-hydroxy-1-(hydroxymethyl) ethyl ester. (Figure 1 and Table 1)

Figure 1: Chromatogram of Amaranthus parganensis.

Table 1: Bioactive compounds determined from Amaranthus parganensis ethanolic extract.

S. No	Compound Name	Molecular Formula	RT Min	% Area	Cas#	Biological Activities Reported
1.	Ethoxy(methoxy)methyl silane	C₄H₁₂O₂Si	7.005	3.446%	~	~
2.	3,7,11,15-Tetramethyl-2-hexadecen-1-ol	C₂₀H₄₀O	25.918	36.724%	102608-53-7	Anti-inflammatory, antimicrobial ³⁰
3.	3,7,11,15-Tetramethyl-2-hexadecen-1-ol	C₂₀H₄₀O	26.246	3.336%	102608-53-7	Anti-inflammatory, antimicrobial ³⁰
4.	3,7,11,15-Tetramethyl-2-hexadecen-1-ol	C₂₀H₄₀O	26.492	9.016%	102608-53-7	Anti-inflammatory, antimicrobial ³⁰
5.	n-Hexadecanoic Acid	C₁₆H₃₂O₂	27.856	8.669%	57-10-3	Anti-inflammatory ³¹
6.	3,4,5-Trimethoxy-2',4'-diaminodiphenylsulfide	C₁₅H₁₈N₂O₃S	28.195	2.621%	24891-45-0	~
7.	Squalene	C₃₀H₅₀	28.964	7.740%	111-02-4	Antidiabetic, Antioxidant, Anti-inflammatory, Antitumor ³⁰
8.	Phytol	C₂₀H₄₀O	29.374	8.148%	150-86-7	Antimicrobial, Anti-inflammatory, Anticancer ³⁰
9.	9-Octadecenoic acid (Z)-, 2-hydroxy-1-(hydroxymethyl)ethyl ester	C₁₂H₄₀O₄	29.979	20.300%	3443-84-3	Antimicrobial, Anti-inflammatory, Anticancer ³²

Characterization of Silver Nanoparticles:

Characterization of nanoparticles helps us to understand the morphology and chemical constituents of nanoparticles²⁹.Here the synthesized silver nanoparticles were characterized via SEM and EDX spectroscopy. From the SEM micrographs (at different magnifications), we can observe a cluster of relatively spherical and non-uniformly distributed silver nanoparticles with a degree of aggregation (Figure 2). The EDX spectra showed the presence of elements such as Ag, C, O, Cl, Na. In EDX pattern the presence of peaks before 4 keV shows the presence of the pure metal ion silver (Figure 3). The SEM micrographs and EDX spectra confirms the presence of silver nanoparticles synthesized from Amaranthus parganensis.

Fig 2: SEM micrographs of silver nanoparticles from ethanolic extracts of Amaranthus parganensis.

Fig 3: EDX Spectra of Amaranthus parganensis.

DISCUSSION:

Amaranthus parganensis is a novel plant whose activities are not known until now. The bioactive compounds that were detected using GC-MS includes phytol; squalene; 9-Octadecenoic acid (Z)-, 2-hydroxy-1-(hydroxymethyl) ethyl ester; 3,7,11,15-Tetramethyl-2-hexadecen-1-ol; 3,4,5-Trimethoxy-2',4'-diaminodiphenylsulfide; N-Hexadecanoic Acid and Ethoxy (methoxy) methyl silane. Many of the compounds that have been identified in our study have shown various types of biological activities. Various studies have already reported the antioxidant, anti-cancer and anti-inflammatory activities of these compounds^30-32. Among these compounds, the following compounds namely 3,4,5-Trimethoxy-2',4'-diaminodiphenylsulfide and Ethoxy(methoxy)methyl silane are those whose activities are relatively unknown. Moreover, through this study we have shown that silver nanoparticles via green synthesis were synthesized using the ethanolic leaf extract and these silver nanoparticles have great potential for therapeutic and medicinal use. We conclude that the plant Amaranthus parganensis has important bioactive compounds and can be utilized for silver nanoparticles synthesis, thereby making it a potential source for medicinal and therapeutic applications in the future.

ACKNOWLEDGEMENT:

The authors are grateful to the management of SRM Institute of Science and Technology for providing the facilities to carry out the research work.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

REFERENCES:

1. Fabricant DS., Farnsworth NR. The Value of Plants Used in Traditional Medicine for Drug Discovery. Environ. Health Perspect. 2001; 109: 69–75.

2. Mahadeva Rao U.S et al. Screening of Phytochemicals and Comparative Antioxidant activity of Leaf and Fruit of MalaysianMengkudu Using Aqueous and Organic Solvent Extracts. Research J. Pharm. and Tech. 2013; 6(9): 1064-1072.

3. Ranjitha Dhevi V.S et al. Potential Medicinal Plants to Treat Leprosy - A Review. Research J. Pharm. and Tech. 2018; 11(2): 813-821.

4. Yuan H et al. The Traditional Medicine and Modern Medicine from Natural Products. Molecules. 2016; 21(5): 559.

5. Chen SL et al. Conservation and sustainable use of medicinal plants: problems, progress, and prospects. Chin Med. 2016; 11(37).

6. Cowan M. Plant products as antimicrobial agents. Clin. Microbiol. Rev. 1999; 12(4): 564–582.

7. Muthukumaran P, Shanmuganathanm P and Malathi C. Antioxidative and antimicrobial study of Aerva lanata. Asian Journal of Biochemical and Pharmaceutical Research. 2011; 2(1): 265-271.

8. Sasidharan S et al. Extraction, isolation and characterization of bioactive compounds from plants’ extracts. Afr. J. Tradit Complement Altern Med. 2011; 8(1): 1–10.

9. Abutbul S et al. Screening of desert plants for use against bacterial pathogens in fish. The Israeli Journal of Aquaculture - Bamidgeh. 2005; 57(2): 71-80.

10. Khan AW et al. Phytochemical analysis and enzyme inhibition assay of Aerva javanica for ulcer. Chemistry Central Journal. 2012; 6(76): 1-6.

11. Peter K, Gandhi P. Rediscovering the therapeutic potential of Amaranthus species: A review. Egyptian Journal of Basic and Applied Sciences. 2017; 4(3): 196-205

12. Das S. Amaranthus parganensis (Amaranthaceae), a New Species from West Bengal, India. Novon: A Journal for Botanical Nomenclature. 2015; 23(4): 406-410.

13. Nana FW et al. Phytochemical composition, antioxidant and xanthine oxidase inhibitory activities of Amaranthus cruentus L. and Amaranthus hybridus L. extracts Pharmaceuticals. 2012; 5(6): 613-628

14. Sharma N, Gupta PC, Rao Ch V. Nutrient content, mineral content antioxidant activity of Amaranthus viridis and Moringa oleifera leaves. Res J Med plants. 2012; 6(3): 253-259.

15. Jimoh MO, Afolayan AJ and Lewu FB. Antioxidant and phytochemical activities of Amaranthus caudatus L. harvested from different soils at various growth stages. Sci Rep 2019; 9: 12965.

16. Amabye T. Evaluation of Physiochemical, Phytochemical, Antioxidant and Antimicrobial Screening Parameters of Amaranthus spinosus Leaves. Nat Prod Chem Res. 2015; 4(1).

17. Ritu Paliwal et al. Elucidation of Free Radical Scavenging and Antioxidant Activity of Aqueous and Hydro-Ethanolic Extracts of Moringa oleifera Pods. Research J. Pharm. and Tech. 2011; 4(4): 566-571.

18. Bharathi V, Vijaya Anand A. Chemical Characterization from GC-MS Studies of Ethanolic Extract of Macrotyloma uniflorum. Research J. Pharm. and Tech. 2016; 9(3): 238-240.

19. Palaniswamy S and Gurusamy K. Free radical scavenging activity of Amaranthus blitum. Pharmacology online. 2010; 3: 625-636.

20. Iordache A et al. Characterization of some plant extracts by GC–MS. Nuclear Instruments and Methods in Physics Research Section B: Interactions With Materials and Atoms. 2009; 267(2): 338–342

21. Suman T, Chakkaravarthi K, Elangomathavan R. Phyto-chemical profiling of Cleistanthus collinus leaf extracts using GC-MS analysis. Research J. Pharm. and Tech. 2013; 6(11): 1173-1177.

22. Priya S, Nethaji S, Sindhuja S. GC-MS Analysis of Some Bioactive Constituents of Diospyros virginian. Research J. Pharm. and Tech. 2014; 7(4): 429-432

23. Kanagavalli U et al. Plant Assisted Synthesis of Silver Nanoparticles Using Boerhaavia diffusa Leaves Extract and Evolution of Antibacterial Activity. Research J. Pharm. and Tech. 2016; 9(8): 1064-1068.

24. Shamalie A and Kalaimathi J. A Comparative Antimicrobial Activity of Annona Squamosa Aqueous Leaves Extract and its Mediated Synthesized Silver Nanoparticles. Research J. Pharm. and Tech. 2016; 9(12): 2228-2233.

25. Krithiga N, Rajalakshmi A and Jayachitra A. Green Synthesis of Silver Nanoparticles Using Leaf Extracts of Clitoria ternatea and Solanum nigrum and Study of Its Antibacterial Effect against Common Nosocomial Pathogens. Journal of Nanoscience. 2015; (1): 8

26. Anket Sharma et al. Phytochemical and Elemental Analysis of Brassica juncea L. Leaves using GC-MS and SEM-EDX. Research J. Pharm. and Tech. 2015; 8(12): 1662-1664.

27. Manimaran T et al. Biosynthesis of Green Nanoparticles from Occimum sanctum and their Characterization. Research J. Pharm. and Tech. 2016; 9(4): 397-400.

28. Forough M and Farhadi K. Biological and green synthesis of silver nanoparticles. Turkish J. Eng. Env. Sci. 2010; 34(4): 281-287.

29. Suresh Dhanaraj P et al. Biosynthesis and Characterization of Silver Nanoparticles from Aspergillus niger and its Antibacterial Activity. Research J. Pharm. and Tech. 2018; 11(12): 5282-5286.

30. Sudha T, Chidambarampillai S and Mohan VR. GC-MS analysis of bioactive components of aerial parts of Fluggea leucopyrus Willd. (Euphorbiaceae). Journal of Applied Pharmaceutical Science. 2013; 3(5): 126-130

31. Aparna V et al. Anti-inflammatory property of n-hexadecanoic acid: Structural evidence and kinetic assessment. Chem Biol Drug Des 2012; 80: 434-9.

32. J Hussein, Hussein and Al-Masudi, Hadi and Hameed, Imad. Study of chemical composition of Foeniculum vulgare using Fourier transform infrared spectrophotometer and gas chromatography -mass spectrometry. Journal of Pharmacognosy and Phytotherapy. 2016; 8(3): 60-89.

Received on 01.02.2021 Modified on 29.04.2021

Research J. Pharm.and Tech 2022; 15(4):1523-1526.

DOI: 10.52711/0974-360X.2022.00253