E-ISSN:2456-3110

Research Article

Vishalyakarani

Journal of Ayurveda and Integrated Medical Sciences

2023 Volume 8 Number 9 September
Publisherwww.maharshicharaka.in

Quantification of Flavonoids & Phenols presents in Tridax procumbens L. (Vishalyakarani) Leaves found in Gandhamardhan hills, Bargarh (Odisha)

Mohanthy S1*, Rautaray K2, Mahesh M3, Nayak S4
DOI:10.21760/jaims.8.9.9

1* Shiva Prasad Mohanthy, Associate Professor, Department Of Dravyaguna Vigyana, Monark Ayurveda Medical College And Hospital, Ahmedabad, Gujarat, India.

2 Kshirabdhi Tanaya Rautaray, Professor, Dept of Samhita Siddhanta Sanskrit, Monark Ayurveda Medical College and Hospital, Ahmedabad, Gujarat, India.

3 Mallipudi Durga Mahesh, Sri Sri Nrusinghnath Ayurved College Research Institute Nrusinghnath, PO Paikmal Bargarh, Odisha, India.

4 Sonali Nayak, Post Graduate Scholar, Department of Kaumarabhritya, KAHERS Shri BMK Ayurveda Mahavidyalaya, Belagavi, Karnataka, India.

Background: Vishalyakarani or Tridax procumbens L. is an Ayurvedic herb under asteraceae family with traditional use in wound healing purpose. It has many pharmacological properties including anti-microbial, anti-cancer, wound healing and anti-septic. Purpose: The present study is a combined study consisting reviewing the anti-microbial lspectrum of Tridax pocumbens L. (Vishalyakarani) and quantification of flavonoids and phenols from its leaves specially from the ganghamardhan hill area, Odisha. This aims to equip researchers a useful data about the chemical contents of T. procumbens. The extract was prepared by using Soxhlet extractor and concentrated by distillation. Phytochemical analysis revealed the presence of flavonoids, phenols in extract and quantification by the high-throughput 96-well plate method. Method: The review process yielded 748 journals and after thorough discussion 81 papers were selected. The plant has many chemical compounds including flavonoids, essential oils, phenols and wide range of pharmacological activities from which we have included the anti-microbial activities. Conclusion: Historically and from Ramayana we have heard about the effectiveness of the Vishalyakarani in wound healing and in present review proves its worth. But there was a lack of quantification of two important phyto contents and its done by high-throughput 96-well plate method.

Keywords: Tridax procumbens, Hot oven method, Soxhlet Extractor, Micro Plate Reader, 96-well plate method

Corresponding Author How to Cite this Article To Browse
Shiva Prasad Mohanthy, Associate Professor, Department Of Dravyaguna Vigyana, Monark Ayurveda Medical College And Hospital, Ahmedabad, Gujarat, India.
Email: 		Mohanthy S, Rautaray K, Mahesh M, Nayak S, Quantification of Flavonoids & Phenols presents in Tridax procumbens L. (Vishalyakarani) Leaves found in Gandhamardhan hills, Bargarh (Odisha). J Ayu Int Med Sci. 2023;8(9):54-58.
Available From
https://jaims.in/jaims/article/view/2766

Manuscript Received Review Round 1 Review Round 2 Review Round 3 Accepted
2023-07-15 2023-07-25 2023-08-02 2023-08-16 2023-08-27
Conflict of Interest Funding Ethical Approval Plagiarism X-checker Note
None declared Nil Yes 22.22

© 2023by Mohanthy S, Rautaray K, Mahesh M, Nayak Sand Published by Maharshi Charaka Ayurveda Organization. This is an Open Access article licensed under a Creative Commons Attribution 4.0 International License https://creativecommons.org/licenses/by/4.0/ unported [CC BY 4.0].

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SP Mohanthy et al. Quantification of Flavonoids & Phenols presents in Tridax procumbens L.

Introduction

Tridax procumbens L. is a famous Ayurvedic herb under Asteracea family and included in binomial nomenclature system by Linnaeus in 1753.[1] It is seen as a dominant weed in many states such as Maharastra, Odisha, Bihar.[2] It is the most potent species among 30 species of the genus Tridax.[3] This herb. T. procumbens have been used from ancient times to treat wounds, skin diseases and to stop blood clotting. It possesses anticoagulant, antileishmanial, antioxidants, anticancer, immunomodulatory agent, insecticidal, anthelmintic cardiovascular, antiseptic, antimicrobial, and insecticidal properties.[4]

Methodology

Review Methodology

An internet based literature search of T. procumbens L was done using Google scholar, Science direct, Google patent, Pubmed, ACS and Mendley desktop databases of plants. Keywords used for literature survey, plant location, and antimicrobial spectrum of T. procumbens L. Undergraduate students of SSN Ayurveda College and Research Institute have participated in this exercise and study conducted in Pharmacognosy lab of SSN Ayurved College and RI, Paikmal, Bargarh (Odisha) and Dept of Biotechnology and Bioinformatics, Sambalpur University, Burla (Odisha). The recommended reporting items for systematic reviews and Meta Analysis (PRISM) guidelines were followed.

Pharmacological studies methodology

Different extracts were studied for their pharmacological potentials and here special emphasis was given for anti-microbial activities both in vivo and in-vitro.

Anti-microbial activities

The anti-bacterial activities with control, MIC/MBC by different methods are mentioned. Various extracts were used such as ether, chloroform, and ethanolic by Agar well diffusion method against B.faecalis, B. subtilis, E. coli, P.aeruginosa .The chloroform extracts was found to be effective against B.faecalis and E.Coli.[5] After going through all these literatures the summery of the anti-microbial activities are reflected in the given table.

 

Review summery of Anti microbial activities of T. procumbens L. (Vishalyakarani)

PartsClass of CompoundMethodsEffectsStandardsDosesReference
LeavesTannins,SteroidsAgarwell diffusionB. subtillisB.faecalisE.ColiAmpicillin  200µg400µg800µg.Christudas et al., (2012)
LeavesFlowersFlavonoidsROS,Lipid peroxidationM. aeruginosAmpicillin25mg/L-100mg/L for 10 daysMecina et.al.,2019
WholePlantFlowersTanninsCarbohydrateDisk diffusionmethodsE.ColiS. typhiS. paratyphi--(Taddei –Rosas-Romero2000)
Fresh flowersEssential oilsTubedilutionassayE.ColiS.typhiS.paratyphiM.segmentis-0.14-0.67mg/LJoshi and Badakar 2012)
Whole plantEssential oilsDisk diffusionmethodS. aureusE.ColiS.typhiK. pneumoniaShigell. flaxineriGentamycin-Mir. S andDar. A 2016
Aerial partEssential oilsAgar wellDilutionmethodMicrospermum fulvum,Microsporumgypsum,GriseofulvinMIC 1.6 to 12.8mg/mLPolicegudret al.,2014
Fresh leavesEssential oilsDisk diffusion S. aureus,S. pneumoniaNorfloxacin-5µg/mLCefepime-10µg/mLGatifloxacin-15µg/mLMIC/MFC 25-100µg/mLManjamlaiEt al.,2012a
LeavesEssential oilsAgar well dilutionCandida albicansAsperagillusfumigatesA.nigerCandida tropicalis-MIF50-500µg/mLManjamlai et al, 2012b.
ArielPartsFatty acidDisk diffusionmethodE. coli,P. mirabilisS. aureusStreptomycinAmpicillin25-10mg/mLAndriana et al., 2019
Leaf-Disk diffusionmethodE. coli,S. aureusS. pyogenesisAmpicillinCephalosporin-T.et al., 2018

 



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SP Mohanthy et al. Quantification of Flavonoids & Phenols presents in Tridax procumbens L.

Morphology

Tridax procumbens has a stem which can reach from to 8-30 inches (20-75 cm) long. The leaves are opposite, pinnate, oblong to ovate, and 1-2 inches (2.5-5 cm) long with cuneate bases, coarsely serrate margins, and acute apexes. The flowers have white rays and yellow disk flowers. They are about 0.4-0.6 inches (1-1.5 cm) wide, and held on a 4-12 inches (10-30 cm) long stalk. Flowering occurs in spring. Fruits are achenes that are dark brown to black in color, oblong, and 0.08 inches (2 mm) long, each with a head of pappus bristles that ovary from 0.12-0.24 inches (3-6 mm) long.[6]

Phytochemical study

Flavonoids

The Phytochemical studies revealed that Tridax procumbens L. is a rich source of Flavonoids, responsible for antioxidant, hepatoprotective, Anticancer, Antibacterial & Wound healing properties. The flavonoids responsible for control the growth of toxin-producing bacteria in plants.[7]

Phenolic Compounds

Tridax procumbens L leaves contents Phenolic compounds having antioxidant properties. The Anti-oxidants are prevent to damage cells by free radicals, the free radicals released during metabolic process of oxidation. The free radicals are reactive oxygen free radicals species (ROS), hydroxyl radicals (OH.), superoxide anion radical (O2), hydrogen peroxides (H2O2) and peroxyl (ROO.). These free radical generates metabolites, which are damage cell membrane DNA. The free radicals are causing carcinogenesis.[8]

Estimation of Phenols and Flavonoids by 96 well plate method

Jaims_2766_01.jpg

Micro Plate Reader

Procedure:

Estimation of phenols:

Reagents for Phenols (Total Phenol Content):

1. Folin reagent 10 % Solution: In the measuring jar Folin reagent is taken 10 ml and adjust 100 ml of volume with the distilled water
2. Sodium Carbonate 5 % solution: In the volumetric flask 5g of Sodium Carbonate is taken and make up with distilled water up to mark in 100 ml.

Procedure: 2 mg of Aqueous Extract is taken in vial. 1.5 ml of Folin reagent is added, 1.5 ml of Sodium Carbonate solution and 2 ml Distilled water is added and made a solution for 96 well plate method solution kept in 37 °C. From this solution 0.1, 0.2, 0.3 ml is added in three wells. The absorbance of the reaction mixture was read at 765 nm using a visible-UV micro plate kinetic reader.[9]

Estimation of Flavonoids

Reagents for Flavonoids (Total Flavonoids Content):

1. Aluminum Chloride 10 % solution: In the volumetric flask 10 g of Aluminum Chloride is taken and make up with distilled water up to mark in 100 ml.
2. Sodium Hydroxide 4 % solution: In the volumetric flask 4 g of Sodium Hydroxide is taken and make up with distilled water up to mark in 100 ml.

Sample: 2 mg of Aqueous Extract is taken in vial. 1.5 ml of Aluminum chloride is added, 1.5 ml of Sodium Hydroxide solution and 2 ml Distilled water is added and made a solution for 96 well plate method solution kept in 37°C. From this solution 0.1, 0.2, 0.3 ml is added in three wells. the absorbance of the reaction mixture was read at from 367 to 510 nm using a visible-UV micro plate kinetic reader.[10]

Result

Estimation of Phenols and Flavonoids by 96 well plate method:

TFC X= Readings obtained 0.420, 0.426, 0.419

Y=0.0098


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SP Mohanthy et al. Quantification of Flavonoids & Phenols presents in Tridax procumbens L.

Amount of TFC of 200 µL contains = = 42.85µg

For 1000 µL contains=  =214.25 (For 2 mg of extract)

For 3.583 gm extract =  = 383.82 mg

TPC X= Readings obtained 0.389, 0.380, 0.386

Y=0.0166

Amount of TFC of 200 µL contains = = 23.43 µg

For 1000 µL contains=  =117.15 (For 2 mg of extract)

For 3.583 gm extract =  = 209.87 mg

Result: The micro plate reader method yielded the following results

Total Phenol Content: 209.87 mg= 5.85%

Total Flavonoids Content 383.82 mg=10.71%

Discussion

The plant extracts have great potential use as antimicrobial compounds against various microorganisms. This plant can be used as antibiotics in various infections. The combined effect of Phenols and flavonoids against various susceptible microbes leads to new choice of treatment for various microorganisms and in future can be used as the drugs of choice for resistant infections. Secondly the plant is found abundantly and the extractions methods are relatively cheap so economically viable.

Conclusion

Review of different journals leads to the conclusion that its leaves are definitely anti-microbial in different concentrations for a varieties of microorganisms. Leaves collected from The Gandhamardhan hill forest contain 10.71 % flavonoids & 5.85 % phenolic compound in aqueous crude which seems marvelous and opens new avenues for further isolation and quantification of other vital phyto constituents.

Future study

Preclinical studies could be done to test its effectiveness in animal models and latter on clinical trials could be planned.


Acknowledgement

We are very much thankful to Department of Bioinformatics & Biotechnology, Sambalpur University, Burla and SSN Ayurveda College and RI, Nrusinghnath, Paikmal (Bargarh) Odisha for allowing us to use the Pharmacognosy lab under Dept of Dravyaguna.

References

1. Chauhan, B.S and D.E Johnson, Ecology of two troublesome asteraceae species of Rain field rice : Chromolanea odorota and Coat Buttons (Tridax procumbens) Johnson Weed. Science, Cambridge, 2008:56:567-573.

2. Chauhan, B.S and D.E Johnson, Ecology of two troublesome asteraceae species of Rain field rice : Chromolanea odorota and Coat Buttons (Tridax procumbens) Johnson Weed. Science. Cambridge, 2008: 56:567-573.

3. Chauhan, B.S and D.E Johnson, Ecology of two troublesome asteraceae species of Rain field rice : Chromolanea odorota and Coat Buttons (Tridax procumbens) Johnson Weed. Science, Cabridge, 2008: 56:567-573.

4. Chauhan, B.S and D.E Johnson,Ecology of two troublesome asteraceae species of Rain field rice : Chromolanea odorota and Coat Buttons (Tridax procumbens) Johnson Weed. Science.Cambridge,2008: 56:567-573.

5. Muthusamy, R.K. Vasu, L Kangaraj, D.Ponnampallam and B. Wilson. Phytochemical screening and evaluation of Antibacterial activities of methanol extract of Tridax procumbens, International Journal of Pharmacy and Biological Science., New Delhi,2013:3(1),521-524.

6. Chauhan, B.S and D.E Johnson,Ecology of two troublesome asteraceae species of Rain field rice : Chromolanea odorota and Coat Buttons (Tridax procumbens) Johnson Weed. Science. Cambridge, 2008: 56:567-573.

7. Bhat, R.S., J.Shankarappa, H.G. Shivakumar,. A review on Pharmacognosy, Pre-Phytochemistry and Pharmacological analysis of Tridax procumbens. Asian Journal of Pharmaceutical Sciences, Tokyo,2007:2(1),11-17.

8. Perez-Gregorio RM, Garcia-Falcon


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MS, Simal-Gandara J, Rodriguez AS, Almeida DPF, Identification and quantification of flavonoids in traditional cultivars of red and white onions at harvest. Journal of Food Compos Anal, Reading (U.K) 2010:23(6)592-598.

9. Perez-Gregorio RM, Garcia-Falcon MS, Simal-Gandara J, Rodriguez AS, Almeida DPF, Identification and quantification of flavonoids in traditional cultivars of red and white onions at harvest. Journal of Food Compos Anal, Reading (U.K),2010; 23(6)592-598.

10. Perez-Gregorio RM, Garcia-Falcon MS, Simal-Gandara J, Rodriguez AS, Almeida DPF, Identification and quantification of flavonoids in traditional cultivars of red and white onions at harvest. Journal of Food Compos Anal, Reading (U.K),2010; 23(6)592-598.


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