INTRODUCTION:

Amaranthus spinosus which is also known as Pig weed is a very common plant found to grow throughout India and other tropical countries¹. The plant is believed to have several medicinal aspects. The leaves of the plant are considered to have properties to treat menstrual disorder, gastroenteritis, gall bladder inflammation, abscesses, arthritis and also snakebite². The tribal people of Kerala boil the leaves without salt and consume for 2-3 days to cure jaundice³.

Taxonomic Classification: Kingdom: Plantae

Division: Magnoliophyta

Order: Caryophyllales

Family: Amaranthacaea

Genus: Amaranthus

Species: spinosus

Figure- 1: Amaranthus spinosus

Various pharmacological published reports have also revealed that the plant has anti-protozoal, anti-inflammatory and analgesic property⁴. A study performed on the fractions obtained from methanolic leaf extract of A. spinosus revealed that four of the fractions obtained amongst 17 fractions showed antimicrobial activity against Staphylococcus aureus⁵.

In recent years, GC-MS and FT-IR has played an important role in pharmaceutical analysis. GC-MS analysis is a breakthrough in analysis of phytoconstituents and structure elucidation of these compounds as they have a sensitivity of detecting compounds as low as 1ng⁶. FT-IR is considered to be the most powerful tool for identifying the types of chemical bonds (functional groups) present in compounds. The wavelength of light absorbed is characteristic of the chemical bond as can be seen in the annotated spectrum, the chemical bonds in a molecule can bedetermined⁷. The issue of antimicrobial resistance is one major problem arising in the scientific advancement of today’s medical world⁸. This situation has compelled the researchers to develop efficient new antimicrobial agents⁹. Plants are rich in secondary metabolites which has a lot biological activity¹⁰. Therefore, this study was undertaken to analyse the various phytochemical constituents found in the ethanolic extract of leaf of A. spinosus using GC-MS and FT-IR which may provide an insight of its use in traditional medicine.

OBJECTIVE OF THE STUDY:

As Amaranthus spinosus traditionally have been found to have many medicinal values therefore this study has been planned with the objectives to find the phytochemicals of the ethanolic leaf extract of the plant and also to identify the bioactive metabolites of the leaf extract through GC-MS and FT-IR analysis.

MATERIAL AND METHODOLOGY:

Collection of plant material:

The healthy fresh leaves of Amaranthus spinosus was collected from Panikhaiti, Assam, India. Voucher specimen has been deposited in the Department of Botany, Gauhati University and has been identified as Amaranthus spinosus L. Family Amaranthaceae Acc. No. 18132 dated 25.04.2016. Collected leaves were washed with water and rinsed with sterile distilled water.

Preparation of extract:

The leaveswere shade dried under room temperature and then ground into a coarse powder. One hundred gram of the powdered material were taken and cold extraction was performed using ethanol as a solvent. The flasks containing the mixture was placed in the water bath for 48 hours and it was stirred at regular intervals. It was then filtered, evaporated and concentrated which provided us the final extract.

Phytochemical Analysis:

The ethanolic extract was subjected to preliminary phytochemical analysis to assess the presence of various phytoconstituents¹⁰

The phytochemical tests were performed by the conventional methods

1) Test for alkaloids: It was performed using the Wagner’s test.

2) Carbohydrate test: It was performed using Molish test and Benedict test

3) Protein identification:Xanthoproteic test was used for identification of protein.

4) Glycoside test:Libermann’s test and Salkowski’s test were performed to determine the presence of glycoside.

5) Tanin test:Test for tannin: About 0.5gram of dried powered sample was boiled in 2ml of water in a test tube and then filtered. A few drops of 0.1% ferric chloride solution were added and observed for brownish green or blue-black colouration.

6) Saponin test:

a) 5ml of aqueous extract was shaken vigorously with 5ml of distilled water in a test tube and warmed. The formation of the stable form was taken as an indication for the presence of saponin. The frothing was mixed with 3 drops of olive oils and shaken vigorously and then emulsion was formed.

b) Foam test: A very little quantity of the extract was taken and mixed with water and shake it vigorously. If foam produced persist for ten minutes it indicate the presence of saponin.

7) Flavonoids:

a) 2ml of dilute sodium hydroxide was added to 2ml of extract. Yellow colour appearance indicates the presence of flavonoids.

b) Flavonoids give dull green/ reddish brown colour on treatment with ferric chloride solution.

c) Alkaline reagent test: To the test solution, add few drops of sodium hydroxide. Intense yellow colour was formed which turn colourless on addition of few drops of dilute HCL indicates flavonoids.

8) Steroids: Salkowski’s test andLibermann test was performed to determine the presence of steroids

9) Terpenoids:Carr- Price Reagent test was performed to determine the presence of terpenoids

10) Phenolic compound test:When 1% aqueous or alcoholic ferric chloride is added,an intense green, purple, blue or black colour is seen in the solution. This procedure is modified by using a fresh aqueous mixture of 1% ferric chloride and 1% potassium ferricyanide.

GC-MS Analysis:

GC-MS analysis of the active ethanol extract of Amaranthus spinosus was carried out by using the GC-MS instrument (Model Clarus 680 GC and amp, Perkin Elmer USA), equipped with a capillary column’Elite-5MS’ having dimensions-length-60m. ID-0.25mm and film thickness-0.25°. The instrument was operated in electron impact mode at ionization voltage (70eV), an injection volume of 1µl was employed at an injector temperature of 280°C. The carrier gas was Helium (99.9% purity) at a flow rate of 1ml/min. The oven temperature was initially programmed at 60°C for 3 mins and then increased at the rate of 6°C/min to 300° for a holding time of 10mins. The identification of compounds from the spectral data was based on the available mass spectral records (NIST-2008 library).

Fourier Transform Infrared Spectrophotometer (FTIR) Analysis:

Dried powder of the ethanolic extract of Amaranthus spinosus were used for FTIR analysis. Translucent sample discs were prepared by encapsulating 10 mg of the dried extract powder into 100 mg of KBr pellet. The powdered sample of Amaranthus spinosus was loaded in FTIR spectroscope, with a scan range from 400 to 4000 cm^-1 with a resolution of 4cm^-1.

Observation and Result:

Table 1: Phytochemical tests of the ethanol extract:

Name of the Test	Qualitative Result
Alkaloid test	Positive
Carbohydrate test	Positive
Protein test	Positive
Glycoside	Positive
Tanin test	Negative
Saponin test	Positive
Flavonoid test	Positive
Steroid test	Positive
Terpenoid test	Negative
Phenolic compound test	Negative

The GC-MS analysis to identify the bioactive compounds present in Amarnthus spinosus was performed. Ethanolic extract was used for the purpose because it was the most competent extractcontaining a large number of volatile compounds which could be detected in it. The distinctive chromatogram of the of the ethanolic leaf extract of A. spinosus is shown in figure1. The analysis separated and identified 78 known compounds belonging to different chemical classes (Figure 2). The major compounds included 4-Piperidinone,2,2,6,6-Tetramethyl, Methyprylon and 2 Hexanol, 2- Methyl. Few other compounds identified in the extract are2-Pentanol,2,3 – Dimethyl, 3-Octanol, 13- Octadecenal,(2), Z,Z-6,28-Heptatriactontadien-2-One, N- Hexadecanoic Acid, Pentadecanoic Acid, Octadecanoic Acid, Tetradecanoic Acid, Tridecanoic Acid. Many of these identified compounds were known to possess many pharmacological activities.

Figure 2: GC-MS chromatograph of ethanolic leaf extract of Amaranthus spinosus

Table2: The major phytocompounds detected in the ethanolic leaf extract of Amaranthus spinosus by GC-MS analysis

Sl No.	Name of the compound	Corresponding Retention Time*	Total Number of times detected
1.	4-Piperidinone, 2,2,6,6-Tetramethyl	19.583, 19.573	07
2.	Methyprylon	19.583, 19.573	06
3.	2 Hexanol, 2- Methyl	10.629	02
4.	2-Pentanol,2,3 – Dimethyl	10.629	02
5.	3-Octanol	10.629	03
6.	13- Octadecenal, (2)	44.258	02
7.	Z,Z-6,28-Heptatriactontadien-2-One	44.258	01
8.	N- Hexadecanoic Acid	40.121	03
9.	Pentadecanoic Acid	40.121	02
10.	Octadecanoic Acid	40.121	02
11.	Tetradecanoic Acid	40.121	02
12.	Tridecanoic Acid	40.121	03

Therefore, we assume that the strong bioactivities exhibited by A. spinosus in this study are correlated to the occurrence of these bioactive compounds in the ethanolic solvent extract. However, further studies on the isolation, characterization and biological evaluation of these identified compounds are necessary to confirm their potential benefits.

Figure 3: FT-IR spectrum report of ethanolic leaf extract of Amaranthus spinosus

DISCUSSION:

Amaranthus spinosus has been used as a plant for medicinal purpose since many years. In a study conducted by Jaya Mathur et.al. 2010, on Pharmacognostic and Preliminary Phytochemical Investigations of Amaranthus spinosus (Linn.) Leaves revealed the presence of flavonoids, phytosterols, glycosides, tannins, phenolic compounds and carbohydrates¹¹. Fernard W. Nana et.al (2012) had studied on phytochemical composition of Amaranthus cruentusL and Amaranthus hybridus L extract. The phytochemical analyses revealed the presence of polyphenols, tannins, flavonoids, steroids, terpenoids, saponins and betalains¹². Our study also found the presence of Alkaloid,Carbohydrate, Protein, Glycoside, Saponin, Flavonoid and Steroid.

Another study that was conducted by Solomon A. Muru et.al. (2018), had performed a GC-MS analysis of aqueous and ethanolic extracts of Amaranthus spinous leaves. In the study, they found the presence of 1,5-methano-1H,7H,11H-furo[3,4-g]pyrano[3,2-b]xanthene-7,15-dione,3,3a,4,5-tetrahydro-8-hydroxy-3,3,11,11-tetramethyl-1,13-bis(3-methyl-2-buten-1-yl)-, (1R,3aS,5S,14aS)- and 7,11-hexadecadienal as the most abundant constituents while the least abundant compounds were the 9-oxabicyclo[6.1.0] nonane and 7-octen-2-one¹³. In another study conducted by Paranthaman R et. Al (2012), they had found the ethanol extract of the plant showed 12 constituents, the major constituents were Isoxazolidine, 5-hexyl-(+)-(peak area 0.47%), 1,2,4-Butanetriol (peak area 0.47%), trinitrate, N-Ethyl-N’- nitroguanidine (peak area 0.47%), Phenol, 2,6-bis(1,1-dimethylethyl)- 4-methyl, methylcarbamate (peak area 2.37%), 2,5-Pyrrolidinedione, 3-ethyl-1, 3-dimethyl-(peak area 0.16%), 2-Piperidinone, N-[4-bromo-n-butyl]-(peak area 0.16%), 3-Hexadecyloxycarbonyl-5-(2- hydroxyethyl)-4-methylimidazolium ion (peak area 0.16%), 2-(3-Oxo- 3-phenylpropyl)-3,5,6-trimethylpyrazine (peak area 0.47%), Pyridine- 3-carboxamide, 4-dimethylamino-N-(2,4-difluorophenyl)-(peak area 1.11%), Phytol (peak area 84.97%), Pseudoephedrine, (+)-(peak area 2.06%), Aspidofractinine-3-methanol (peak area 7.12%)⁽¹⁴⁾. Varalakshmi Thanikachalam et.al.conducted the GC-MS analysis of Amaranthus viridus. In the study they found the presence of Benzoic acid, 2-[(trimethylsilyl)oxy]-, trimethylsilyl ester (24.09%) is found as the major compound and forty nine minor compounds such as Nonamethyl, Phenyl-, Cyclopentasiloxane (6.10%), (+,-)-3.beta.-(acetyloxy)-3-ethynyl-1,2,3,4,4a. beta.,12a. (5.67%), [Bis (trimethylsilyl) methyl] diphenyl phosphine $$ Phosphine (5.59%), Benzoic acid, 2-[(trimethylsilyl) oxy]-, trimethylsilyl ester (5.32%), 1,2-Diphenyl tetramethyldisilane $$ Disilane,1,1,2,2-tetramethyl-1,2-diphenyl (4.98%), Benzophenone, 2-(trimethylsiloxy)- (CAS) Trimethylsilyl ether of O-hydroxybenzophenone (3.50%), 7-(P-Chlorophenyl)imino-6-(P-tolyl)-1,3-dimethyl-2,4 dioxo-1,2,3,4,6,7- hexahydro pyramid [4,5-d] pyramidine (3.14%), 4-.alpha.,20-dimethyl-3-.beta.-dimethyl-.. (2.84%), Silane, [1,3,5-benzenetriyltris (oxy)] tris [trimethyl- (CAS) Phloroglucinol tris MS (2.76%), 2-Isopropyl phenol- Trimethylsilyl- Ether (2.73%), 4H-1-Benzopyran-4-one, 2-(2,6-dimethoxyphenyl)- 5,6-dimethoxy- (CAS) Zapotin $$ Flavone (2.46%), (Z)-1-[(1’,1’-dimethylethyl)diphenylsilyl]- 3-trimethylsilyloxyprop-1-ene (2.37%), Prosta-5,10,13- trien-1-oic acid, 15-[[(1,1-dimethylethyl) dimethyl ethylsily] oxy]-9-oxo- (2.19%), Silane, trimethyl (triphenylethenyl)- $$ (2.18%) and the remaining compounds peak area ranged from 1.84% to 0.16%¹⁵. The present study on the ethanolic extract of Amaranthus spinosus we found the presence of similar compounds like 4-Piperidinone,2,2,6,6-Tetramethyl, Methyprylon and 2 Hexanol, 2- Methyl, 2-Pentanol,2,3 – Dimethyl, 3-Octanol, 13- Octadecenal,(2), Z,Z-6,28-Heptatriactontadien-2-One, N- Hexadecanoic Acid, Pentadecanoic Acid, Octadecanoic Acid, Tetradecanoic Acid, Tridecanoic Acid.

In a study conducted by Ratul Kumar Das et.al (2012) on the FT-IR analysis of the ethanolic leaf extract of A. spinosus, showed the presence of characteristic bands for several functional groups. IR peaks for hydroxyl (-OH), aromatic amines (-C₆H₅NH₂), aliphatic amines (R-NH₂), carbonyl (>C=O), C-H and C=C (benzene) functional groups were observed at around 3378, 1118, 1380, 1040, 1694, 2848 and 1604 cm⁻¹ respectively¹⁶. Another study performed on the FT-IR analysis of Methanolic extract of Curcuma caesiaby Muthukumaran Pakkirisamy et.al. in 2017, confirmed the presence of N-H, O-H, C=C, C-H, C-O and CH₃ functional groups¹⁷. In a similar study performed on the ethanolic extract of Amaranthus spinosus, the presence of similar functional groups like OH stretch, C-H Alkane, Aromatic rings, Phenol or tertiary alcohol, Aromatic ether, Aliphatic fluoro compounds were confirmed.

CONCLUSION:

From this study it may be concluded that Amaranthus spinosus is a plant of great medical importance. Confirmation on the presence of phytochemical compounds like Alkaloid, Carbohydrate, Protein, Glycoside, Saponin, Flavonoid and Steroid determined the importance of the plants. Also the presence of bioactive compounds like 4-Piperidinone,2,2,6,6-Tetramethyl, Methyprylon and 2 Hexanol,2- Methyl, 2-Pentanol, 2,3 – Dimethyl, 3-Octanol, 13- Octadecenal, (2), Z,Z-6,28-Heptatriactontadien-2-One, N- Hexadecanoic Acid, Pentadecanoic Acid, Octadecanoic Acid, Tetradecanoic Acid, Tridecanoic Aciddetermines the medicinal value of the plant because the same compounds have been found to be present in many other medicinal plants. FTIR spectra showed the presence of the functional group in the extract. The results obtained is a significant finding in this present study.

ACKNOWLEDGEMENT:

I would like to thank the management of Guwahati Biotech Park, IIT, Guwahati for providing us the infrastructure. I would also like to thank the management of Assam down town University for providing the infrastructure and seed grant for the research work. This is apart of PhD work of Miss Sangita Boro.

CONFLICT OF INTEREST:

None.

REFERENCE:

1. Choudhury A. Evaluation of physiochemical and phytochemical parametersof Amaranthus spinosus, International Research Journal of Pharmacy. 2012; 3(10):210-211; https://irjponline.com/admin/php/uploads/1456_pdf.pdf

2. Kawade, R. M., Ghiware, N. B., Sarje, S. K., andVadvalkar, S. M. A pharmacognostic and pharmacological review: Amaranthus spinosus. World Journal of Pharmaceutical Research. 2013; 2(6):99-110; https://doi.org/10.29252/nirp.bcn.8.6.503

3. Jhade, D., Ahirwar, D., Jain, R., Sharma, N. K., and Gupta,S. A pharmacological review: Amaranthus spinosus. Research Journal of Pharmacognosy and Phytochemistry. 2009; 1(3):169-172; https://rjpponline.org/HTMLPaper.aspx?Journal=Research%20Journal%20of%20Pharmacognosy%20and%20Phytochemistry;PID=2009-1-3-6

4. Boro, S., Bharadwaj, B., and Unni, B.G. Prickly Amaranth (Amaranthus spinosus Linn.): a Review on Phytochemistry and Pharmacological Activities. American Journal of Pharmatech Research, 2016; 6(5):81-92.http://ajptr.com/archive/volume-6/october-2016-issue-5

5. Majumdar, J., Boro, S., Bharadwaj, B., and Unni, B.G. Phytochemical analysis and Isolation of Compounds from Methanolic Leaf Extract of Amaranthus spinosus and its antimicrobial effect against Staphylococcus aureus. World Journal of Pharmaceutical and Life Sciences. 2017; 3(7): 100-103.https://www.wjpls.org/home/article_abstract/638

6. Saravanakumar, K., Adaikala, R., and Umaiyambigai, D. GC-MS and FTIR Profiling of leaves methanol extract from the Pleiospermiumalatum (Wall. ex Wt. and Arn) Swingle Rutaceae family. J Phytopharmacol. 2016; 5: 201-204. http://www.phytopharmajournal.com/Vol5_Issue5_06.pdf

7. Ashokkumar, R., and Ramaswamy, M. Phytochemical screening by FTIR spectroscopic analysis of leaf extracts of selected Indian medicinal plants. International Journal of Current Microbiology and Applied Sciences. 2014; 3(1): 395-406. https://www.ijcmas.com/vol-3-1/R.Ashokkumar%20and%20M.%20Ramaswamy.pdf

8. Moellering Jr, R. C., Graybill, J. R., McGowan Jr, J. E., and Corey, L. Antimicrobial resistance prevention initiative—an update: proceedings of an expert panel on resistance. The American Journal of Medicine. 2007; 120(7): S4-S25.doi: 10.1016/j.amjmed.2007.04.001

9. Ayoub, I. M., El-Shazly, M., Lu, M. C., and Singab, A. N. B. Antimicrobial and cytotoxic activities of the crude extracts of Dietesbicolor leaves, flowers and rhizomes. South African Journal of Botany. 2014; 95: 97-101. https://doi.org/10.1016/j.sajb.2014.08.012

10. Rekha, R., Hemachander, R., Ezhilarasan, T., Keerthana, C., Saroja, D. L., Saichand, K. V., and Abdullah, M. G.. Phytochemical analysis and in-vitro antioxidant activity of Mimosa pudica Lin., leaves. Research Journal of Pharmacy and Technology. 2010; 3(2): 551-555. https://rjptonline.org/AbstractView.aspx?PID=2010-3-2-43

11. De Fátima, A., Modolo, L. V., Conegero, L. S., Pilli, R. A., Ferreira, C. V., Kohn, L. K., and De Carvalho, J. E. Styryl lactones and their derivatives: biological activities, mechanisms of action and potential leads for drug design. Current Medicinal Chemistry. 2006; 13(28): 3371-3384 https://doi.org/10.2174/092986706779010298

12. Mathur, J., Khatri, P., Samanta, K. C., Sharma, and Mandal, S. U. Pharmacognostic and preliminary phytochemical investigations of Amaranthus spinosus (Linn.) leaves. International Journal of Pharmacy and Pharmaceutical Sciences. 2010; 2(4): 121-124.https://www.researchgate.net/publication/282696803_Pharmacognostic_and_preliminary_phytochemical_investigations_of_amaranthus_spinosus_LINN_leaves

13. Nana, F. W., Hilou, A., Millogo, J. F., and Nacoulma, O. G. Phytochemical composition, antioxidant and xanthine oxidase inhibitory activities of Amaranthus cruentus L. and Amaranthus hybridus L. extracts. Pharmaceuticals. 2012; 5(6): 613-628.https://doi.org/10.3390/ph5060613

14. Mamuru, A. S., Kaigamma I, and Khan, E. M. Preliminary Phytochemical Screening, and GC-MS analysis of Aqueous and Ethanolic Extracts of Amaranthus spinosus leaves. Journal of Natural Products and Resources. 2019.; 5(2): 213-215.https://doi.org/10.30799/jnpr.071.19050201

15. Paranthaman, R., Praveen, K. P., and Kumaravel, S. GC-MS analysis of phytochemicals and simultaneous determination of flavonoids in Amaranthus caudatus (Sirukeerai) by RP-HPLC. J. Anal. Bioanal. Tech. 2012; 3(147): 2020.https://doi.org/10.4172/2155-9872.1000147

16. Thanikachalam, V., and Jayaraj, I. A. Phytochemistry of Amaranthus viridis: GC-MS Analysis. Int J Cur Res Rev. 2021; Vol 13(07), 162.https://doi.org/:10.31782/IJCRR.2021.13713

17. Das, R. K., Gogoi, N., Babu, P. J., Sharma, P., Mahanta, C., and Bora, U. The synthesis of gold nanoparticles using Amaranthus spinosus leaf extract and study of their optical properties; Advances in Materials Physics and Chemistry. 2012; 2(4); https://doi.org/10.4236/ampc.2012.24040

18. Pakkirisamy, M., Kalakandan, S. K., and Ravichandran, K. Phytochemical screening, GC-MS, FT-IR analysis of methanolic extract of Curcuma caesiaRoxb (Black Turmeric). Pharmacognosy Journal. 2017; 9(6).https://doi.org/10.5530/pj.2017.6.149

19. Sureka, A., Dhanalakshmi, R., Sharmila, C. M., and Devi, R. C. Comparative Analysis on Significance of Purification Process of Monosilai in Siddha medicine using Modern Instrumental Techniques: Standardization Approach. Asian Journal of Pharmaceutical Analysis. 2019; 9(2): 99-104.https://doi.org/10.5958/2231-5675.2019.00019.X

20. John, C., Idris, M., Sudhakar, P., Ghosh, P., Varshney, K. M., and Shukla, S. K. Determination of Clonazepam in chocolate using High Performance Liquid Chromatography and further confirmation by Gas Chromatography-Mass Spectrometry. Asian Journal of Research in Chemistry. 2011; 4(5): 761-765. https://ajrconline.org/AbstractView.aspx?PID=2011-4-5-21

21. Anandan, A., Eswaran, R., Doss, A., Sangeetha, G., and Anand, S. P. Investigation for bioactive compounds of Aervalanata (L.) Juss. ex Schultes. Asian Journal of Research in Chemistry. 2012; 5(4): 469-471. https://indianjournals.com/ijor.aspx?target=ijor:ajrc&volume=5&issue=4&article=005

22. Rajabudeen, E., Ganthi, A. S., and Subramanian, M. GC-MS Analysis of the Methanol Extract of Tephrosia villosa (L.) Pers. Asian Journal of Research in Chemistry. 2012; 5(11): 1331-1334. https://www.researchgate.net/publication/310468515_GCMS_ANALYSIS_OF_THE_METHANOL_EXTRACT_OF_TEPHROSIA_SPINOSA_L_F_PERS_RAJABUDEEN

23. Kulkarni, A., Jan, N., and Nimbarte, S. GC-MS, FT-IR and NMR Spectroscopy Analysis for Metabolome Profiling of Thyme Oil. Asian Journal of Research in Chemistry. 2013; 6(10): 945-949. https://ajrconline.org/HTML_Papers/Asian%20Journal%20of%20Research%20in%20Chemistry__PID__2013-6-10-12.html

24. Jose, B. E., and Selvam, P. P. Identification of Phytochemical Constituents in the Leaf Extracts of Azimatetracantha Lam using Gas Chromatography-Mass Spectrometry (GC-MS) analysis and Antioxidant Activity. Asian Journal of Research in Chemistry. 2018; 11(6): 857-862. https://ajrconline.org/HTML_Papers/Asian%20Journal%20of%20Research%20in%20Chemistry__PID__2018-11-6-5.html

25. Ruhila, A., Srivastava, A., Yadav, P., Galib, R., and Prajapati, P. K. Preliminary GC-MS profiling of Bhallataka Oil. Asian Journal of Research in Chemistry. 2021; 14(2): 108-110. https://doi.org/10.5958/0974-4150.2021.00019.5

26. Suman, T., Chakkaravarthi, K., and Elangomathavan, R. Phyto-chemical profiling of Cleistanthuscollinus leaf extracts using GC-MS analysis. Research Journal of Pharmacy and Technology,(2013); 6(11): 1173-1177. https://www.researchgate.net/publication/258249267_Phytochemical_profiling_of_Cleistanthus_collinus_leaf_extracts_using_GC-MS_analysis

27. Priya, S. S., Sharmili, A. S., Anbumalarmathi, J., and Umamaheswari, K. Evaluation of Phytochemical Constituents, In vitro Antimicrobial, Antioxidant, FT-IR and GC-MS Studies of Leaves of Solanum torvum, Rhizome of Acorus calamus and Whole Plant of Mollugo pentaphylla. Research Journal of Pharmacy and Technology. 2017; 10(2), 592-600. https://doi.org/10.5958/0974-360X.2017.00117.2

28. Gunalan, G., Kumar, M. S., and Sangeetha, N.. Preliminary Phytochemical Analysis and In Vitro Oxidant Scavenging Activity of Rosemary officinalis. Research Journal of Pharmacy and Technology. 2011; 4(10): 1588-1590. https://www.researchgate.net/publication/287686476_Preliminary_phytochemical_analysis_and_in_vitro_oxidant_scavenging_activity_of_Rosemary_officinalis

Received on 09.08.2021 Modified on 13.06.2022

Research J. Pharm. and Tech 2023; 16(8):3685-3690.

DOI: 10.52711/0974-360X.2023.00606