INTRODUCTION:

D. confragosa (Bolton) J. Schröt. locally known as “lakri phootu”, in the Indian context, but it is globally known as a Blushing bracket, due to the presence of shades in pink or mauve in the upper surface of the cap. It’s having a tough nature of cap, slow-growing fungus, most occurring in dead or dying trunks. Previously, it is called thin-maze polypore, even though some different varieties in altered in mazes position. This is a saprobic fungus and occurs in temperate regions in universally forest biodiversity¹. It is an inedible nature for human or pet animals, but also attention due to having a nutritional and medicinal property, therefore in modern science presented as a good resource for antimicrobial and antioxidants potent materials.

Some other bioactive components were reported by the contemporary researcher from worldwide. D. confragosa is the richness for lipid and the fatty acid such as 32.9% neutrallipids, 53.8% phospholipid, 13.3% glycolipids and 20.1% total lipids (mg/g dry weight)^2,3. Also reported that the hydroxyl fatty acid content of D. confragosa, its having a various concentration of total fatty acids, such as 0.02% 7-hydroxy-8, 14-dimethyl-9-hexadecenoic acid and 0.01% 7-hydroxy-8, 16-dimethyl-9-octadecenoic acid. Similarly, some triterpenes family also found, alike 3α-carboxyacetoxy-24-methylene-23-oxolanost-8-en-26-oic acid, 5α, 8α-epidioxyergosta-6, 22-dien-3β-ol (ergosterol peroxide) and 3α-carboxy acetoxy quercinic acid⁴. Its containing various antioxidant components that are capable of scavenging to free radicals, prevented from oxidative damage in living organisms regulates to normal physiological effects⁵. The diabetic preventing mechanism, there is two important carbohydrate hydrolyzing key enzymes like as α-amylase and α-glycosidase, which is the presence in mushrooms, nowadays they are fascinating applying in the postprandial blood glucose level controls and act as potent anti-diabetic medication⁶. The conventional inhibitors of these enzymes also caused the wide side effect, i.e. flatulence, diarrhea and abdominal bloating⁷. The aim of this study was to investigate extracts preparation by using various polar solvents. Then determine the in vitro antioxidant capacity of extract, through using the various procedures such as DPPH scavenging, reducing power assay, total phenolic and hydrogen peroxide scavenging etc. Furthermore, analyzed the anti-diabetic effectiveness of D. confragosa extracts.

MATERIALS AND METHODS:

Sample Collection:

The fresh fruiting body of polypore mushroom D. confragosa collected from dried Shorea robusta tree trunk from Charama forest, Kanker district (Chhattisgarh) India, at the session of October 2015. These collected samples were kept in polythene bags and packed loosely, also given the specific code. Initial identification was made on the basis of critical observations of the specimens and examination of relevant literature⁸. The identified mushroom germplasm deposited to Mushroom Biotechnology Laboratory, at School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur (Chhattisgarh) India for further analysis.

Standard and Reagent:

EDTA, Ascorbic acid, Gallic acid, Ethanol, Methanol, 1,1-Diphenyl-2-picryl-hydrazyl-hydrate (DPPH), Ferrozine, Sodium phosphate buffer, Potassium Ferricyanate [Fe(CN)6], Ferric chloride (FeCl₃), Trichloroacetic acid (C₂HCl₃O₂), Folin-ciocalteu (FC) reagent, Sodium carbonate (Na₂CO₃) Phosphate buffer, Sodium Chloride (NaCl) Phenol red, Sodium Hydroxide (NaOH), Fehling’s solution, Chloroform, Sulfuric acid (H₂SO₄), Glacial acetic acid, HCl, Alcohol, Ammonia and also deionized water. The entire chemical reagents were purchased from Sigma Aldrich Bangalore and HiMedia Pvt. Ltd. Mumbai India.

Extracts Preparation Methods:

This procedure was carried out according to the method with some slight modification^9,10.Initially, the mushroom sample of D. confragosa was cut into small pieces; a sample of 25 g of D. confragosa was extracted by stirring with 100 mL distilled water at 25 ^oC at 150 rpm, for 24 h, and boiled at water bath (100 ⁰C) for 2 h. Similary performed for methanol and ethanol extractions. For the methanolic and ethanolic extractions a sample of 25 g of D. confragosa was extracted by stirring with 250 mL methanol (purity 99.8%) and ethanol (purity 94.0%) at 25 ⁰C at 150 rpm, for 24 h respectively and subsequently filtered through Whatman no. 4 paper. The residue was extracted for the next 48 h. The collected extract was dried at room temperature. All the extracts were dissolved in 5% DSMO solution in distilled water and kept in refrigerated conditions (4^◦C temp.) until further tests. The extracted dissolved residue was coded individually such as hot water extracted (CHE), methanolic extracts (CME) and ethanolic extract (CEE) respectively.

Yield Extract Measurement:

The % yield extract was calculated using the following Equation¹¹: yield (%) = (weight of extract in g/weight of defatted dried mushroom in g) ×100.

Antioxidant Assay:

A. 2,2-diphenyl-1-picrylhydrazyl radical (DPPH)-Scavenging Assay:

The free radical-scavenging activities of mushroom extracts were determined by using the method with minor modification¹². Briefly, 3 mL of each mushroom extract with different concentrations (20 -100 µg/mL) were mixed with 1 mL of the methanolic solution containing with DPPH (0.1 mM). The mixture was shaken vigorously and left to stand for 30 min in the dark at room temperature and the absorbance was then measured with a quartz glass cuvette) at 517 nm against a blank using a UV-visible spectrophotometer . The low absorbance of the reaction mixture illustrated a high free-radical-scavenging activity¹³. The absorbance of radical without DPPH was used as a control and EDTA was used as a standard. The graph was plotted in which X-axis showed concentration (µg/mL) and Y-axis showed the scavenging effect (%). The percentage of inhibition was calculated according to the formula:

(Ao – A1)

DDPH scavenging effect (% ) = ^{--------------------}× 100

(Ao )

Where Ao was the absorbance of the control reaction and A1 the absorbance in the presence of the sample of mushroom extract respectively. The activity was also expressed as the inhibition concentration at 50% (IC₅₀), the concentration of solution for the test required to obtain 50% of radical scavenging capacity.

B. Reducing Power Assay:

The reducing power of mushroom extracts was determined according to the modified method^14,15. Different concentrations (20-100 µg/mL) of mushroom extract (25 µl) were mixed with sodium phosphate buffer (25 µl, 200mM, pH 6.6) and 25 µl of 1 percent (v/v) potassium ferricyanide. The mixture was incubated at 50^◦C for 20 min and 25µl of 10 percent (v/v) trichloroacetic acid was added to the mixture and centrifuged at 3000 rpm for 10 min. After the centrifugation, the upper layer of solution (100 µl) was mixed with deionized water (100 µl) and 10 µl of 0.1 percent (v/v) ferric chloride. The absorbance of the test extract was measured at 700 nm (ELISA Reader, Microscan MS 5608A). Ascorbic acid was used as the positive control. The graph was plotted against the standard in which X-axis showed concentration (µg/mL) and Y-axis showed anti-radical scavenging activity (%) and a higher absorbance indicates a higher reductive capability. The percentage of reducing power was calculated by an equation

(Control OD – Sample OD)

% of Reducing power assay = ----------------------------------- ×100

Control OD

C. Total Phenolic determination:

Phenolic determination estimated by using the method ^15,16with minor modification. The different concentration of mushroom extracts 50 µl and 50 µl folin-ciocalteu phenol reagent was taken and mixed it and after that 3 minutes saturated Sodium carbonate solution of 50 µl was mixed, its make up to 300 µl with distilled water. Solution kept in dark room for 90 minutes then absorbance was recorded at 725 nm by using 96 well microtitre plate containing by ELISA Reader (Microscan MS 5608A). The Gallic acid was used as a standard. A graph was plotted against the standard in which the X-axis showed con. (µg/mL) and Y axis showed inhibition (%).The percentage of inhibition was calculated according to these formula:

(Control OD – Sample OD)

% of Total phenolic antioxidant capacity=--------------------------- ×100

Control OD

14 15 14 16

D. Hydrogen peroxide scavenging:

This hydrogen peroxide scavenging capacity measured according to method¹⁶ with minor modification. In this method, different concentrations of mushroom extract 20 µl were added to 20 µl of (0.002% ) hydrogen peroxide. Then added 160 µl of 0.1 M phosphate buffer and 100 mM NaCl were added, the reaction mixtures were incubated for 10 min at 37^◦C. Then 1 ml 0.2 mg/mL phenol red was added. After 15 min 10 µl of 1M NaOH was added and absorbance were measured by ELISA Reader (Microscan MS 5608A) at 610 nm. The graph was plotted against standard in which X-axis showed concentration (µg/mL) and Y-axis showed inhibition (%). The percentage of hydroxyl radical scavenging calculating formula is given below.

(Control OD – Sample OD)

% of Hydroxyl radical scavenging assay = -------------------------- ×100

Control OD

E. Antidiabetic Property assay (α-amylase inhibitory activity):

This antidibatic assay performed according to techniques^17,18 with minor modification.The CHE , CME and CEE extracts were diluted (50 μl) and 50 μl of 0.02 mol/L Sodium phosphate buffer (pH 6.9 with 0.006 mol/L NaCl) containing α-amylase (EC 3.2.1.1; 0.5 mg/mL) were incubated at 37 °C for 05 min. Then, 100 μl of 1% starch solution in 0.02 mol/L Sodium phosphate buffer (pH 6.9 with 0.006 mol/L NaCl) was added to the reaction mixture. Thereafter, the reaction mixture was incubated at 37 °C for 03 min and stopped with 100 µl of dinitrosalicylic acid (DNSA).The mixture was then incubated in a boiling water bath at 100^◦C for 8 min, and cooled at room temperature. Then the absorbance was measured at 450 nm in an ELISA Reader (Microscan MS 5608A).

F. Statistical analysis:

All the experiments were performed in three parallel triplicates. The values are presented as mean +SE (n=3). The statically difference among the groups was analyzed by using a one-way variance analysis test by using SPPS programme (SPSS 16.00). The significant levels were defined at P<0.05. The graphical data were created in Microsoft Excel version 7.0.

RESULTS:

Effects of Solvent on Extraction Yield:

D. confragosa, which was collected from Charama forest at session October, 2015 and on the basis of morphology characterization, identification was done with the critical analysis of feature with standard references as per described^19,20. Extraction of defeated powder sample was carried out with employing various polar solvent (CHE), methanol (CME) and ethanol (CEE) respectively. The extraction process might be expected that removal of containing undesirable fatty substance from the prepared powder. This process also improved the polarity of phenolic compounds that increase their yields in extracts individually. The yield of mushroom extracts percentage depicted in Table 1. Among the extracting medium, hot water was the best extracting solvent for D. confragosa, with the percentage recovery of 50.00 (g/dried weight). This was followed by ethanol with the achieved rate of 03.65 %, while methanol extracted gave only 02.70 % respectively.

Table: 1.Yield of Mushroom extracts by different solvents

Extraction solvent	Yield of Extract (g/100 dry weight)
Hot Water 60 °C (CHE)	50.00
Methanol (CME)	02.70
Ethanol (CEE)	03.65

DPPH Scavenging Activity:

DPPH method is fast and quick assay for evaluating to radical scavenging activity. The result of DPPH radical scavenging activity of the various extracts of mushroom and ascorbic acid were depicted as the ways of increasing percentage with the increased concentration manner, shown in Fig. 1. The result indicates that the methanolic extracts (CME) showed the strongest DPPH radical scavenging activity (94.39±0.01), which was higher than the positive control (Ascorbic acid) at (100 µg/mL). Among the other solvent fractions, the ethanolic extracts (CEE) gave moderate scavenging activities (76.60±0.04), and the water extract (CHE) showed the very lesser scavenging activity (55.48±0.00) at 100 µg/mL respectively. For the standards (Ascorbic acid) was recorded (91.03±0.09) at maximum scavenging efficiency at the concentration of extracts (100 µg/ mL). However, the methanolic solvent has extracted maximum secondary metabolites contains from samples, its having a stronger scavenging role. The increased order to DPPH scavenging activity of different mushroom extracts was represented in following CHE< CEE< CME respectively.

Fig. 1. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity of polar solvent extracts of D. confragosa (Bolton) J. Schrot. All values are reported as the mean ± standard error (n=3)

The IC₅₀ values were presented in Table 2. The methanolic extracts showed good inhibitory activity (IC₅₀= 17.05 µg/mL), followed by ethanolic extracts (IC₅₀=18.07 µg/mL) and water extracts depicted very lesser activity.

Reducing Power Assay:

Reducing power of different extracts were compared with standard ascorbic acid. The reducing capacity of mushroom extracts was found to be similar for all used extracts, in order to the values increased with increasing the concentration of extracts gradually.

The results of reducing power of different mushrooms from different extracts were plotted in Fig. 2. The present experiment recorded that standard (Ascorbic acid) was gave lower reducing power, scavenging variation showed with the taken extracts mushroom samples. Among the all polar solvents extracts, the water extracts gave a highest reducing power (84.12±0.00), followed by ethanolic extracts (75.27±0.01) and methanolic extract gave (75.18±0.01) at (100 µg/mL). The highest reducing power shown by water extracts (CHE), it might be having a rich concentration of total phenolics and flavonoids contents, The orders of reducing the ability of different extracts are following: ascorbic acid<CME<CEE< CHE etc. The IC₅₀ value was presented in Table 2. According to the IC₅₀value increased the order of the different mushrooms from the different sample are following: CHE < CEE < CME< STANDARD. In this study, CHE showed the lowest IC₅₀=16.77µg/mL value.

Fig. 2. Reducing power assay of polar solvent extracts of D. confragosa (Bolton) J. Schrot. All values are reported as the mean ± standard error (n=3)

Total Phenolic determination:

Generally, the concentration of phenolic in the extracts expressed as μmoles of GAEs per mg of extracts. In the present studied, the total phenolic content was depicted in Fig. 3. The highest scavenging activity was found for hot water (CHE) extracts 77.29±0.03, followed methanol extracted showed (CME) 76.14±0.01, moderate scavenging capacity but ethanolic extracts (CEE) had given (72.82±0.03) at (100 µg/mL), thus its very poor phenolic scavenging activity. The standard (Gallic acid) gave antioxidant potency, contrary less than in water extracts (CHE) mushroom (72.82±0.03). These are represented also in the increased manner of phenolic content scavenging efficiency of mushrooms are the following: CEE< STANDARD< CME< CHE etc. These results also supported our research finding. The IC₅₀ values of mushroom extract shown in Table 2. The IC₅₀ of the standard sample was very lesser (16.19 µg/µL) then other polar solvent extracts samples. The methanolic extracts (CME) showed very potential and significant IC₅₀ values, in comparison to ethanolic and hot water extracts respectively.

Fig. 3. Total phenolic antioxidant capacity of polar solvent extracts of D. confragosa (Bolton) J. Schrot. All values are reported as the mean ± standard error (n=3)

Hydrogen Peroxide Scavenging:

The H₂O₂ reducing capacity of compound or extract might serve as indicators of its potential antioxidant capacity. From the analysis, we can conclude that the scavenging effect of mushroom extracts on H₂O₂ radicals increased with the increased concentration manner. The results analyzed depicted in Fig. 4. The water extracts (CHE) showed (99.32±0.03) highest H₂O₂ scavenging activity rather than methanolic (86.85 ±0.03), and ethanolic extracts (CEE), (68.21±0.00) at 100 µg/mL. The increasing order of the mushroom extracts is: CEE< CME< CHE etc. The IC₅₀ value was presented in Table 2. The water extracts showed (IC50= 15.63 µg/mL) strong scavenging capacity, comparison to another polar solvent extracts methanolic extracts (19.32) moderate and ethanolic extracts (81.42) very lesser effectiveness respectively.

Fig. 4. Hydroxyl radical scavenging activity of polar solvent extracts of D. confragosa (Bolton) J. Schrot. All values are reported as a mean ± standard error (n=3)

Antidiabetic Assay:

The result of this studied showed that D. confragosa mushroom extracts inhibited α-amylase in a dose-dependent manner (in the range of 20.00-100.00 μg/mL). The α- amylase inhibiting activities are shown in Fig. 5. The in vitro antidiabetic study by α-amylase inhibition assay using hot water (CHE), methanolic (CME) and ethanolic (CEE) of mushroom. These studied showed similar antidiabetic activity. The maximum of α-amylase inhibition were recorded for CEE (33.69), then CME (33.11) showed moderate inhibition activity and CHE (32.78) showed slightly less inhibitory activity at maximum concentration (100 µl /mL) respectively. The IC₅₀ values obtained were as follows CME (34.63); CHE (37.18) and CEE (48.19) all in (μg/mL). However, assessment of the extracts IC₅₀ (extracts concentration causing 50% enzyme inhibition) values reveals that significant difference did exist among the IC₅₀ of the three extracts, with CHE having a higher inhibitory activity than the other two other extracts, while the CEE have the least inhibitory activity. It could be noted from the Table 2, that the IC₅₀ value is lowest in CEE followed by CHE and the highest value was recorded in CME. The lower the IC₅₀ value the more effective the inhibitory activity. Thus the extract of CME is more effective in terms of inhibitory activity.

Fig. 5. Inhibition of α-amylase determination of polar solvent extracts of D. confragosa (Bolton) J. Schrot. All values are reported as the mean ± standard error (n=3)

DISCUSSION:

Antioxidant compounds are significant bio-product of biological systems; therefore it is a major objective to analysis and evaluations of effectiveness from the wild mushroom. The antioxidant analyzed properties with three polar solvents extracts were evaluated by in vitro mechanism, including DPPH assay, reducing power assay, total phenols determination assay, and H₂O₂as well as antidiabetic properties etc.^21,22. The previous research found that no single solvents considered as standard then use two different solvents for different phytoconstituents conditions²³.The present studied recorded hot water extract gave maximum yielding, among to other used solvents. Similarly, work reported ²⁴, they obtained highest recovery value for Ganoderma resinaceum, in methanol-ethanol combination extraction medium. Furthermore, they were recored the yielded, for methanol-ethanol (08.10%), followed by ethanol had (07.85%) and methanol had (6.40.%) the least recovery percentage. Similary we have obtained for methanolic and ethanolic extracts yielding.

Antioxidant measurement DPPH is base assay mechanism. Because DPPH is a free radical molecules, it has been frequently used for scavenging free radical from food components.It is a stable molecule, to become stable diamagnetic molecules it will accept an electron or hydrogen radical. The current study had shown effectiveness scavenging ability for all polar solvent with increasing concentration manner. The similar results were reported by another coworker²⁵. They found the scavenging activity of DPPH radical of ethanolic extracts of Hypsizigus marmoreus mushroom, they achieved a slightly higher capacity of 75.5%, but also a much a higher concentration (5 mg/mL). In addition, the water extracts of D. confragsoa showed scavenging capacity 36.8% at 5 mg/mL²⁴. The radical scavenging capacity was found to exhibit 50% of the inhibition values (IC₅₀ value) at the extract concentration for methanolic extract (17.05), followed by ethanolic (18.07), standard (ascorbic acid) (19.24%) and lesser capacity was recorded for water extract (19.88). They have recorded that methanolic extracts showed very good free radical scavenging capacity²⁶. They found the maximum activity for Ganoderma lucidum (95.36±0.02%) recorded at 1mg/mL concentration. The 50% inhibitory activity (radical scavenging capacity) was also expressed as the inhibition concentration at (IC₅₀) value. IC₅₀determination is an essetntial parameter for scavenging analyze. The present study found that for D. confragosa ethanolic extracts gave (IC₅₀=18.07 µg/mL), that showing potent scavenging ability. Similar work also presented²⁷. They reported water extract gave (0.015±0.007 mg/mL) . Our obtained values, which indicate that higher antioxidant capacity extracts, comparison to the previously reported . Some another coworker²⁸ reported that the P. ostreatus ethanol extract showed strongly neutralized the DPPH radical with the IC₅₀ being 69.30 µg/mL.

Another antioxidant analyzed the process. Here, ferrous ion is commonly present in food systems; it is suggested as an effective pro-oxidant component and phenolic compounds have the ability that can chelate pro-oxidant metal ion, thus preventing free radical formation. Thus the possible to total phenolic contents responsible for break the free radical chain by donating an electron to stabilize and terminate easily radical chain reaction²⁹. Aslo another researcher reported that the reducing power capacity of D. confrasoa (0.442±0.06) and for standard (Ascorbic acid) (1.192±0.3) at 0.2 mg/mL²⁷. Clitocybe odora showed 0.94 mg/mL and Coprinus comatus showed 0.72 mg/mL IC₅₀value³⁰. Forever the reducing power of the present investigated mushroom extract was higher than previously reported, for same species due to the possible region might be affected to geographical habitat, affected to constituent nature of secondary metabolites and its reducing power capacity.

The antioxidant activity of biological materials was well correlated with the content of their phenolic compounds³¹. For the reason that phenolic compounds are a large group of secondary plant metabolites which played a major role in the protection of oxidation processes³². Therefore numerous studies have been conclusively demonstrated that mushrooms also contain many phenolics which are important phyto constituents they preserve the scavenging ability³³. In addition, phenolic compounds exhibit a wide range of biological effects including antibacterial, anti-inflammatory and antihyperglycemic³³.

It has been reported that phenolic compounds are suggested to be powerful chain-breaking antioxidants. Also, it does possess scavenging ability due to their hydroxyl groups. The similar result was reported by other coworker²⁹, they found the highest amount of total phenolic compounds in mushroom extracts for L. edodes (36.19±0.59), V. volvacea (22.97±0.29), P. eous (20.31±0.56) and P. sajor-caju (16.46±0.67) respectively. Among to these tested mushroom, L. edodes was presented the highest contents of total phenolics (36.19 mg GAE/g dw). Another associates also found 19.51, 24.71 and 16.2mg/mL IC₅₀values for A. bisporus, P. dryinus and L. nuda respectively³⁴. The findings are supporting to our finding but mushroom species differ. Correspondingly, other colleague reported that the mushrooms contain more polar constituents which dissolved with more polar extractant³⁵. Therefore it can be interpreated that the higher contents of total phenolics found in the mushroom extracts could be considered indicative of higher antioxidant ability of water extracts. The hyderogen peroxide scavenging mode for analyzed to free radical molecules. Therefore, H₂O₂ plays an important role as the radical-forming intermediate in the production of ROS molecules. Thus, H₂O₂ is considered as one of the main inducers of cellular aging and could attack many cellular energy-producing systems³⁶. H₂O₂ scavenging capacity of various mushrooms had analyzed , Hypsizygus ulmarius (cap), H. ulmarius (stipe), A. bisporus (cap), A.bisporus (stipe), Calocybe indica (cap) and C. indica (stipe) showed 2.920, 2.995, 0.908, 2.770, 0.993 and 3.252mg/mL at IC₅₀value respectively³⁷. Our achieved result is more significant, comparison to previously reported. For the α-amylase inhibitory analyzed for in vitro determination of antidiabetic potency. The present studied dealt with three various polar extracts of mushroom. All the extracts were showed efficient antidiabetic activity in dose- dependent inhibitory process. The present study finding is supported by other researchers. They reported that 2, 4-Thiazolidinedione derivatives demonstrated efficiently substitute as antidiabetic property³⁸. Similarly, derivatives might be present in D. confragosa, therefore the present studied examined that the polar solvent had shown well in vitro antidiabetic activity. Similary work done by other researchers they analyzed the in vitro antidiabetic inhibitory aqueous extracts for two varieties of ginger³⁹; same related research evidence also reported⁴⁰ , they found α-amylase activity antidiabetic activity for Calocybe indica mushroom. This finding is the support to our results.

CONCLUSION:

Present obtained results revealed that the antioxidant study strongly suggested that the extract of hot water, methanolic and ethanolic extracts of D. confragosa has significant antioxidant activity, also illustrated the measurement methods of antioxidant compounds, which could serve as an easily accessible item of natural rich antioxidant food which may enhance the immune system against oxidative damage or it may be utilized as a potential source of therapeutic agent.

In the antidiabetic assay, the inhibition of saccharides hydrolysable enzymes by the D. confragosa mushrooms in a dose-dependent manner. Hence, the antidiabetic effect of the all extracted solvent of D. confragosa was determined, it might be attributed to the inhibitory activity of the hydrolysable enzymes thereby retarding the digestion of carbohydrate to delay the rise in blood glucose. The taken solvent showed good inhibitory activity for anti-diabetic activity. So, these mushroom is the best natural resources for making to medicine as well as provide to the nutritive constituent for human beings instead of chemical synthetic material drugs etc.

ACKNOWLEDGEMENT:

The authors are thankful to Chhattisgarh Council of Science & Technology, Raipur for providing to Research Project (Project Sanction No. 722/CCOST/MRP/2015, dated 23/07/2015). We are also obliged to Head, School of Studies in Biotechnology for providing necessary facilities during the study.

CONFLICT OF INTEREST:

The authors declare that there are no conflicts of interest.

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Received on 14.07.2018 Modified on 17.08.2018

Research J. Pharm. and Tech 2018; 11(12): 5623-5630.

DOI: 10.5958/0974-360X.2018.01020.X

Mushroom samples	DPPH (µg/mL)	Reducing power (µg/mL)	Total phenolic determination (µg/mL)	Hydrogen scavenging (µg/mL)	Antidiabetic (µg/mL)
CHE	19.88±0.02	16.77±0.02	92.99±0.04	15.63±0.03	37.18±0.12
CME	17.05±0.03	25.53±0.03	45.36±0.03	19.32±0.02	34.63±0.14
CEE	18.07±0.02	19.79±0.02	89.680.01	81.42±0.02	48.19±0.16
Standard	19.24±0.01	63.04±0.02	16.52±0.02	18.18±0.01	10.47±0.02