Insulin Plant Research Articles

Novel characterisation of Curvularia geniculata associated with leaf blight on Costus igneus in India

Costus igneus Nak, commonly referred as insulin plant or spiral flag, is a perennial herb belonging to the family Costaceae. Owing to its ability to regulate the blood sugar level, it is considered as a boon plant for diabetic patients, hence popularly known as the “sugar plant”. Although few foliar diseases have been documented on C. igneus, they generally occur in minor frequency under Indian conditions. However, in May 2022, a moderate to severe incidence of leaf blight was observed on insulin plants at University of Agricultural Sciences, GKVK, Bangalore, and nearby medicinal gardens. The affected leaves exhibited light brown lesions coalesced to form papery blight, gradually leading to complete drying of leaves. The current study aimed to identify, characterize, and document the fungal pathogen responsible for the disease, with a focus on its morphological, molecular, and biological characteristics to ensure accurate identification. Additionally, the study sought to determine optimal conditions for pathogen growth and sporulation to support the development of effective disease management strategies. The associated fungus was isolated, purified and identified as Curvularia geniculata through morphological and multigene phylogenetic analysis, using internal transcribed spacer (ITS) and large sub unit (LSU) regions of rDNA, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Biological characterization revealed that cornmeal agar (CMA) at 30 °C and pH of 7 were ideal for better growth and abundant sporulation. These findings, highlighting the emergence of C. geniculata as a novel fungal pathogen infecting C. igneus in India, are crucial for developing effective disease management strategies to safeguard the medicinal properties of the plant.

Novel derivatives of Costus igneus towards potentiality against diabetes mellitus receptors: ADME/Tox profiling, Computational Docking, and Molecular Dynamics Simulation study

Costus igneus has been practiced to alleviate a range of human ailments and the substantial vision was to explore potential bio-active compounds of Costus igneus to better comprehend the underlying mechanism in the management of diabetes. In-silico approach using AutoDock and Desmond, the anti-diabetic efficacy of discovered phytochemicals was evaluated. The best-docked were chosen for simulation studies. The molecular dynamic modelling showed that the complexes of ligands with receptors were stable during a time of 100 ns, which further confirmed and enhanced their significant binding affinity. According to docking scores and simulation studies, these phytocompounds could be targeted to garner efficient preventive medications for diabetes from a biological source using contemporary technology, and further in vivo and in vitro research of these compounds may be encouraged in the search for a potent treatment for the disease.

A Comprehensive Study on the Micropropagation of Costus igneus: Media Composition, Growth, and Development

Costus igneus, commonly known as insulin plant, is an ornamental plant valued for its spiral ginger- like foliage and medicinal properties. Micropropagation is an efficient method for rapid multiplication of Costus igneus. This study aims to determine the optimal media composition and culture conditions for in vitro propagation of C. igneus. Axillary bud explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of plant growth regulators including auxins (indole-3-acetic acid, indole-3-butyric acid and naphthaleneacetic acid) and cytokinins (6- benzylaminopurine and kinetin). Microshoots cultured on MS + 1.5 mg/L BAP showed the highest shoot proliferation rate (98%) and maximum number of shoots per explant (12.6). Elongated microshoots were rooted on half-strength MS medium supplemented with different auxins. The highest rooting percentage (95%) and maximum number of roots per shoot (9.8) were observed on medium containing 2.5 mg/L IBA. The regenerated plantlets were acclimatized and successfully transferred to pots with 80% survival rate. Morphological and phytochemical analysis showed no significant differences between the in vitro propagated and mother plants. This study demonstrates the potential of micropropagation for large scale production of quality planting material of C. igneus. Further studies on genetic and epigenetic stability are recommended to validate this protocol for commercial applications.

Green synthesis of m-LaVO4 nanoparticles using Costus igneus leaves extracts and its photocatalytic and electrochemical applications

Green synthesis of m-LaVO4 nanoparticles using Costus igneus leaves extracts and its photocatalytic and electrochemical applications

Zinc nanoparticles mediated by Costus pictus leaf extract to study GC-MS and FTIR analysis

Plants have been used to treat illnesses from the dawn of time, and the term “medicinal plants” has taken on special significance. Because of their pharmacological potency, these plants have continued to be used as a source of medicines and other natural goods worldwide. Costus pictus, Commonly known as flaming costus (Costus igneus Nak), is a South-central American plant recently brought to India. It belongs to the Costaceae family. It is called the “insulin plant” in India because of its alleged anti-diabetic qualities. The methods utilized in this study are GC-MS, FTIR and SEM analysis. The results revealed that GC-MS shows the presence of different compounds: fatty acids, terpenoids, Alkanes, Carboxylic acid and aromatic compounds. The FTIR analysis depicts various functional groups comprising alkanes, alkenes, alkynes, carboxylic acid, aromatic compounds, and halo compounds. SEM analysis depicts the shape and size of nanoparticles, and the size of nanoparticles is 77nm – 113nm. Further research should be done on the characterization of Zn nanoparticles, metabolomics analysis and pharmacological activities.

Optimised herbal transdermal patch of ethnobotanical plant Costus igneus N. E. br. Developed through response surface methodology

Costus igneus (Family: Costaceae) has been used by the aboriginal humans since long through oral route. In our laboratory, we used biodegradable polymer for release control of this transdermal patch. This transdermal patch could be affixed in the lower abdominal region which releases intraperitoneally after principal meals, twice a day; which could be terminated instantly by removing the patch externally. Our objective is Formulation, evaluation and statistical designing of a Transdermal patch containing ethnobotanical plant Costus igneus. As Transdermal Patch reduces can be removed when termination of treatment is necessary and it also has the capability to bypass the different chemical changes that a oral pill has to go through. The Macro-physico evaluation parameters were calculated. The folding endurance was in between of 7 to 16, the maximum value was of the FOPT, the Tensile strength was relatively similar in all the formulations. FOPT showed the highest of all. In percentage elongation test the FOPT showed good resilience, thickness was relatively acceptable for all the formulations. % moisture content and %moisture uptake was acceptable in all of the formulations showed a good and satisfiable range. The drug content was calculated according the total quercetin content taking pure quercetin API as the reference FOPT had the highest drug content of 82.5%. The Response Surface Curve predicted the optimised formulation and the desirability score also gives us our optimised patch. The Kinetics for the optimum by the optimization of transdermal patch was observed in all the different type of plots. The FOPT had a good linearity of 0.938 in regression co-efficient with Korsmeyer-Peppas Plot . It explains the release of drug’s material through a polymeric matrix system while also considering Non-Fickian drug release mechanism. The different Response Surface Curve allows us to predict from the software that the FOPT Formulation is the Gem of Ten. This would produce beneficial blood-sugar content reduction. If we can commercialise this Herbal Transdermal Patch then, it can be an innovation remedy for this chronic disease.

In vitro antioxidant activity and hypoglycemic efficacy of the leaf, stem, and rhizome extracts of <i>Costus igneus</i> nak (Costaceae) in Alloxan Induced Diabetic Rats

Costus igneus has a folk reputation in West Africa for the treatment of diabetes. Nevertheless, from our investigations, the scientific validation of this folkloric claim has not been properly documented. The study was designed to evaluate the antidiabetic potentials and in vitro antioxidant capacity of the leaves, stems and rhizomes of C. igneus using ethanol as the extraction solvent. Phytochemical screening of the extracts was performed using standard analytical procedures. Saponins, tannins, flavonoids, alkaloids, and cardiac glycosides were detected in the leaves, stems, and rhizomes of C. igneus. The median lethal dose (LD50) of the leaf, stem, and rhizome extracts in mice were 2958. 04, 1936.49, and 5000 mg/kg, respectively. The treatment of alloxan-induced diabetic rats with the extracts of the leaves, stems, and rhizomes caused a significant reduction (P<0.05) in fasting blood glucose levels (44.13, 56.21, and 61.12%), respectively in acute study, and 76.85, 69.95, and 79.63% reduction in prolong treatment (2 weeks). The leaf and rhizome extract also demonstrated good antioxidant activity on 2, 2-diphenyl-1-picrylhydrazyl radicals (61 and 62% inhibition, 100 μg/mL) and ferric reducing capacity (absorbance, 0.541 and 0.459, 100 μg/mL), respectively. The leaves, stems, and rhizomes of C. igneus exhibited good hypoglycaemic activity which supports their folkloric claims in the management of diabetes mellitus. The rhizomes extract showed the least toxicity and the most effective in the reduction of fasting blood glucose levels. This is the first comparative evaluation of the leaves, stems, and rhizomes of C. igneus in a study. The findings suggest the use of the rhizomes as a better substitute than the leaves popularly employed in herbal preparations or a combination of all the plant parts for an effective herbal treatment. The antioxidant capacity of this plant extracts may also be involved in the inhibition of oxidative processes implicated in diabetic complications.

Formulation and Evaluation of Anti-Diabetic Tablet from Insulin Plant (Costus Igneus)

In this research article we prepare tablet formulation of Costus igneus plant i.e., Insulin plant by its ethanolic leaves extract and mixing with suitable excipients. Extract was prepared by collecting Costus igneus plant leaves and go for shade drying for 4-7 days, after drying its crushed by mill to form powder and taken for extraction. After extraction we prepare powder for taking different chemical qualitative test and laboratory flow properties such as; (Angle of repose, Loose bulk density, Tapped bulk density , Compressibility index) and after checking flow properties and then punch tablet by direct compression method we take two forms of tablet formula changes in main API one is extract and other is only powder and go for evaluation study such as; ( Color ,Weight variation Test ,Hardness, Friability, Disintegration Time, Thickness)

Micro-qualification by estimation of leaf constants of the ethnobotanical plant Costus igneus N.E. Br by optical microscopical analysis

Costus igneus N.E.Br, traditionally known as Insulin Plant, is an ethnobotanical perennial plant used by aboriginal people for treatment for Hyperglycaemia, or abundancy of sugar in body. In modern studies, researchers have found that the leaves of Costus igneus N.E. Br to be effective especially for Type-I Diabetes. In addition to its’s Anti-Diabetic property it has various other properties like Anti-Oxidant, Anti-Urolithic, Anti-Proliferative, Anti-Inflammatory and Anti-Lipidemic property. The Isolated Pharmacologically active moiety is a Flavonoid called Quercetin. A traditional method of identification of a plant species is by estimation of its Leaf Constants. Just like fingerprints, each plant has its own unique identification in terms of leaf constant. The fresh leaves of the plant were collected from Insulin plant of Bengal School of Technology, Hooghly, Herbal Garden and a Herbarium was prepared. The Herbarium was then authenticated and identified from Botanical Survey of India, Shibpur, and West Bengal. The authenticated plant was collected for fresh leaves and the Leaf Constants that were calculated were Stomatal Number, Stomatal-Index, Palisade Ratio, Vein-Islet Number and Vein-Termination Number. The leaf constants were observed and shown in table.

Micro-qualification by estimation of leaf constants of the ethnobotanical plant Costus igneus N.E. Br by optical microscopical analysis

Micro-qualification by estimation of leaf constants of the ethnobotanical plant Costus igneus N.E. Br by optical microscopical analysis

IN VITRO ASSESSMENT OF NATURAL HERBAL EXTRACTS FOR ANTIMICROBIAL ACTIVITY

Objective: Herbal medicine is still the mainstay of about 75-80% of the whole population, and the major part of traditional therapy involves the use of plant extract and their active constituents against infectious pathogens. Objective: The study aimed to investigate in vitro antibacterial activity of extracts from some medicinal plants against the most common microbial pathogens including MDR bacteria.
 Methods: The processing of plant materials was performed with the washing, drying and grinding of collected plant materials. The plant extracts were prepared by mixing 10 g of powder to 150 ml of ethanol solvent for 5 h at room temperature and sonicated for 15 min; for prepared test samples under laboratory conditions, the air-dried samples were mixed with the respective solvent (1:15 w/v) for 72 h at room temperature with occasional and then filtered through Whattman filter paper No.1. The obtained extract was freed from the solvent by evaporation under reduced pressure and then resuspended in the appropriate solvent to make the solution of known concentration of 10-50 mg/ml. The extract was stored at 4 °C in airtight glass bottle for the antibacterial assay using the Agar-well diffusion method. Ciprofloxacin was used as a control antibiotic.
 Results: The growth of K. pneumoniae, Proteus mirabilis, P. aeruginosa and E. coli were inhibited better with the plant extract Tinospora cardifolia leaves than ciprofloxacin antibiotic. Coagulase-negative Staphylococci was inhibited greatly with Costus igneus leaf extract. The growth of Enterococcus faecalic was inhibited significantly with Tridax procumbens leaf extract than ciprofloxacin. 
 Conclusion: The present study indicates Tridax procumbens, T. cordifolia and Costus igneus methanolic leaf extracts were showed strong antimicrobial activity against all the tested cultures. They were rich in primary and secondary constituents. Most of the biologically active phytochemicals were present in methanolic extract. The tested plant extracts were more efficient than standard antibiotic ciprofloxacin used in the current study.

Detailed study on Costus igneus plant for its medicinal importance - A Review

A medicinal shrub called Costus Igneus may have a miraculous treatment for diabetes. The leaf of this medicinal plant, which is a member of the Costaceae family, strengthens the beta cells in the pancreas, which helps the body produce more insulin. This is why it is known as the "Insulin Plant" in India. There is a strong belief that eating the leaves of this plant can control blood glucose levels and diabetics, according to earlier research. For antidiabetic activities, the experts have used various extracts from plant parts or specific phytochemicals. This review provides the scholar with information on numerous activities on various diseases in addition to just antidiabetic activities

In vitro regeneration through organogenesis in Costus igneus–an important herbal insulin plant

Costus igneus is an important high-valued herbal insulin plant. An efficient protocol for in vitro plant regeneration through organogenesis from leaf and nodal explants was standardized. Explants were incubated in Murashige and Skoog (MS) medium amended with diverse levels of plant growth regulators (PGRs). Optimum frequency (68.25%) of adventitious shoot regeneration was obtained from nodal segments cultured onto MS medium containing 2.0 mg/l of 6-benzylaminopurine (BAP) with an average number of 5.0 shoots per explant. Further, MS medium containing 1.5 mg/l 2, 4- dichlorophenoxyacetic acid (2, 4-D) and 1.0 mg/l Thidiazuron (TDZ) produced a higher frequency of callus induction (55.22%) from leaf explants. Optimum shoot regeneration response (66.0%) through indirect organogenesis was observed from leaf-derived callus on MS medium fortified with BAP (2.0 mg/l) and kinetin (0.5 mg/l) with 6 shoots per callus with mean shoot length of 8.0 cm. The shootlets obtained from node and leaf cultures were rooted with a frequency of (63.52%) on MS medium with 1.0 mg/l indole-3-acetic acid (IAA). Plantlets obtained through organogenesis were successfully acclimatized in the greenhouse and field conditions. There were no significant differences among frequency of somatic plants regenerated through either direct or indirect organogenesis. The present study reports on successful plant regeneration through direct and indirect organogenesis in C. igneus. The regeneration protocols discussed here can be used effectively for large-scale multiplication, germplasm survival, pharmacological and genetic manipulation research.

Biopesticide Activity of Costus Igneus -A Preliminary Study

Costus igneus plant was tested for potential biopesticide use against D. melanogaster larvae as a model organism. Aqueous and methanol extracts were prepared using fresh leaves of C. igneus. Methanol extracts experiential more effective with LC50 (50% lethal concentration) at 2.71% whereas aqueous extract recorded 1.25 folds higher than methanol with LC50 at 3.37%. The LC50 values were statistically significant (p < 0.05). Results explain 92.4% and 83.2% competence of methanol and aqueous extracts of C. igneus against D. melanogaster larvae correspondingly. The preliminary results proposes C.igneus plant as a potent source of biopesticide.

EVALUATION OF IPIL-IPIL (Leucaena leucocephala) SEED GUM AS CO-ENCAPSULATING AGENT FOR TARGETED AND CONTROLLED DELIVERY OF POWDERED INSULIN PLANT (Chamaecostus cuspidatus)

Type 2 Diabetes Mellitus (T2DM) is the most common non-communicable disease in the Philippines, characterized by increased blood glucose levels brought by low insulin production or insulin resistance. Chamaecostus cuspidatus is a medicinal plant known for its glucose-lowering property. The controlled release of the C. cuspidatus leaves must be achieved to maximize its antidiabetic property. Leucaena leucocephala is an endemic tree in the Philippines, having its seed as a source of possible co-encapsulating material for drug delivery. Galactomannan, as the main component of the isolated seed gum, can be used as a substitute for an effective moderate drug release to its intended site. Thus, this study evaluates the drug release property and stability of seed gum as a co-encapsulating agent for targeted and controlled delivery of the C. cuspidatus leaves. The encapsulation process of the C. cuspidatus leaves was done through the extrusion method. The stability of the encapsulation was evaluated through in vitro gastrointestinal simulation analysis and was examined using differential scanning calorimetry (DSC), Fourier transform infrared spectrometer (FTIR), and field emission scanning electron microscope (FE-SEM) to verify its surface morphology. The capsules were observed to fully disintegrate at the fed state (pH 5.4) of the simulated gastrointestinal conditions, which is the target site. The fingerprint on the FTIR spectra of the encapsulated drug presented indicates the successful incorporation of the powdered leaves inside the encapsulating material. Morphological micrographs have shown that the resulting capsules were fairly in spherical, having a size of approximately 3.8 mm. Ridges and pores are also present on the surface of the capsules for their immediate disintegration and hydration. Therefore, the L. leucocephala seed gum can be a potential candidate as a co-encapsulating material suitable for effective, targeted, and controlled delivery of C. cuspidatus leaves for maximum antidiabetic benefits.

The Review on Insulin Plant: A Plant of Ayurvedic Used (Costus Igneus Plant)

Costus Igneus (insulin plant) traditionally used medicinal herb which is native to southeast Asia. The plant has been recently introduced into india and it is grow as ornamental plant in south India. India insulin plant contain various phytochemicals constituent llike steroid, alkaloid , flovonoid, Triepene, glycoside and saponins. Its leaves are been used as dietary supplement in treatment of diabetes mellitus. The catchphrase of the plant is: a leaf a day keeps diabetes away” various pharmacological activities include anti diabetic effect, antiproliferative potential, antimicrobial activity Antiurolithiatic property anti-inflammatory potential it effect on learning and memory ,antioxidant activity neuroprotective role hypolipidemic activity etc. The present review article attempt explore various medicinal properties of costus igneus (insulin plant )for research purpose and its suitable formation development in feature for the welfare of mankind.

Phytochemical Screening, GC-MS and FTIR analysis of Bioactive Compounds in Methanolic Leaf Extracts of Costus igneus (CIMLE) as a Natural Source of Drug Lead Molecules for Next-generation Drug-design, Development and Therapeutics

Diabetes mellitus (DM) is a metabolic disorder that affects people of all ages. Increased prevalence of diabetes worldwide has led to the development of synthetic drugs to provide an interim solution to the ill effects of the issue. However, such drugs, although effective as antihyperglycemic agents, are accompanied by significant side effects, costly, and inaccessible to majority of people in remote in underdeveloped countries. Local medicinal plants have been used by such indigenous people through ages to treat such complicate ailments due to their ease of availability and GRAS nature. As diabetes continues to become prevalent, health care practitioners are considering PBNPs as a potential source of antidiabetic drugs due to their high potency and fewer side effects. To better understand the mechanism of action of medicinal plants, their active phytoconstituents are being isolated and investigated thoroughly. Phytochemical screening followed by GCMS analysis of methanolic leaf extracts of Costus igneus depicted the presence of 19 bioactive compounds viz., Bicyclo [3.1.1]heptane, 2,6,6- trimethyl-; 2-Pentadecanone, 6,10,14-trimethyl-; phthalic acid; 3,7,11,15-Tetramethyl-2-hexadecen- 1-ol; 5,9,13-Pentadecatrien-2-one, 6,10,14- trimethyl; 2(3H)-Furanone,dihydro-5-methyl-5- pentyl; Hexamethyl-2,6,10,14,18,22- tetracosahexaen-1-yl]-6-methox.; Oxalic acid, cyclohexyl isohexyl ester; 6-Octadecenoic acid; Phytol; 4,8,12,16-Tetramethylheptadecan-4- olide; 5-methyl-2,3-dihydro-1H-indole; Bis(2-ethylhexyl) phthalate; Adamantane-1-carboxylic acid methyl ester; Benzopyran-6-ol,3,4-dihydro-; Benzoquinoline; gamma-Tocopherol; dimethyl-5,5'-diphenyl-1H,; 1H-benzo[b]furo[2,3-f]indole that may be ADMET predicted for potential biomedical application in the treatment of diabetics and long term complications associated with the disease. 
 Keywords: Costus igneus; CIMLE; GCMS; Phytochemical Screening; Antidiabetic Leads; NDDT; Antidiabetic Medicinal Plant

Biosynthesis of ZnONP Using Chamaecostus cuspidatus and Their Evolution of Anticancer Property in MCF-7 and A549 Cell Lines.

The ZnO nanoparticle synthesis using the leaf part of Chamaecostus cuspidatus was characterized using UV–Vis spectrophotometry, IR, XRD, DLS, FESEM, EDX, TEM, AFM and XPS. The MTT assay was used to examine the cytotoxicity activity against lung epithelial and breast cell lines, and the IC50 value was determined. The presence of ZnO nanoparticles, which range in size from 200 to 800 nm, was confirmed by the absorption peak at 350 nm. The median particle size was 145.1 nm, and the ζ -the potential was −19.45 mV, showing that ZnONP is stable. Zinc, carbon, and oxygen contribute to the elemental composition of ZnONP, as determined by EDX analysis. MTT assay was used to investigate in vitro cytotoxicity in MCF-7 and A549 cell lines. The cytotoxicity activity IC50 value was determined to be 30 μg/mL for the A549 cell line and 37 μg/mL for the MCF-7 cell line.

Comparative evaluation of antimicrobial activity of newer herbal disinfectant formulations on hospital microbial flora

Background: The world is now focusing on use of herbals because they are eco-friendly and less toxic on human health and environment Objectives:To evaluate antimicrobial activity of newer herbal disinfectant formulations on hospital microbial flora Methods: Herbal disinfectants were prepared with medicinal plant leaf extracts of Tridax procumbens, Withania somnifera, Tinospora cordifolia, Costus igneus and Nepata cataria. Hospital microbial flora was isolated by rubbing and rotating sterile swabs from targeted spots. Extract formulations were tested for the antimicrobial activity against five isolatedbacterial strains i.e., Escherichia coli, Pseudomonas aeruginosa, Enterobacter sp., Enterococcus faecalis and Staphylococcus aureus using Kirby-bauer disc diffusion method. Results: Extracts exhibited antimicrobial activity against all tested microorganisms. Among the five different herbal disinfectants, Tinospora cordifolia showed good antimicrobial activity against Escherichia coli, Enterobacter and Enterococcus faecalis with 27mm, 20mm and 14mm of zone of inhibition respectively. Costus igneus showed high antimicrobial activity to Pseudomonas aeruginosa with 22mm of zone of inhibition. The herbal disinfectant formulations in the present study were reported effective results against hospital microbial flora which cause nosocomial infections Conclusions: Herbal disinfectants Tinospora cordifolia, Costus igneus, Withania somnifera exhibits antimicrobial activities against MDR bacterial pathogens isolated from hospital wards.

Effect of synchronized sound waves in the form of Indian Classical Ragas on Phytochemical analysis of Chamaecostus cuspidatus (Nees & Mart.) C. Specht & D. W. Stev

Synchronized sound waves in the form of Indian classical ragas have a remarkable impact on the living beings. According to various studies, Indian Classical music has been confirmed to encourage plant growth positively and it is observed that the sounds from metal-rock, rock, rap, pop, and monotonous sounds have a negative impact on plant growth. The present research work was aimed at finding the impact and effect of the synchronized sound waves and their frequencies in different strings and closed-pipe Indian classical instrumental music displayed through various Ragas, viz: Raga- Kedar (flute), Raga- Kedar (santoor), Raga-Rageshree (sitar), Raga-Bhairavi (flute), Raga- Shree (Sarangi), Raga- Milan kiTodi (Sarod), Raga- Ramkali (Sitar) on phytochemical analysis in the leaves of Insulin (Chamaecostus cuspidatus). The standard deviation values in all of the pure solvents indicates that the data were spread over a wider range of values, which further showed that the treated plants have higher concentration phytochemicals when compared to that of the untreated samples. The total phenol content was found to be 207.66±0.33 mg/GAE per gram extract and 113.33±3.33 mg/GAE per gram extract in case of treated and untreated plant samples of Chamaecostus cuspidatus respectively. Similarly, the total flavonoid content was found to be 37.66±0.33 mg QE/g extract and 26.66±0.33 mg QE/g extract in case of treated and untreated plant samples respectively and the total tannin content was found to be 843±1.99 mg TAE/g extract and 503±0.57 mg TAE/g extract in case of treated and untreated plant samples respectively.