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Amanita Caesarea and A 124 Nibha Gupta. et al. / Vol 4 / Issue 3 / 2014 / 124-129. International Journal of Innovative Drug Discovery e ISSN 2249 - 7609 Print ISSN 2249 - 7617 www.ijidd.com NUTRITIVE PROPERTIES AND ANTIOXIDATIVE ACTIVITY OF AMANITA CAESAREA AND A. LOOSII WILD EDIBLE MUSHROOMS FROM ODISHA Sushri Shanta Tripathy, Ashutosh Rajoriya and Nibha Gupta* Regional Plant Resource Centre, Bhubaneswar -751015 Odisha, India. ABSTRACT The aim of this study to determine the nutritional , mineral and antioxidant properties of two species of mushrooms Amanita caesaria and Amanita loosii, collected from dry deciduous forest of Odisha India. These edible mushrooms were analysed in terms of macro and micronutrient, mineral contents and antioxidant properties through various biochemical and spectrophotometrical analysis. Antioxidant properties were determined by evaluating the presence of phenolic content, ascorbic acid, carotenoids and flavonoids. The average protein content ranged 3.13 – 8.66 mg/gm. And carbohydrate ranged 23.61-26.00 g/100g were found. Both mushrooms have exhibited 72-73 % DPPH scavenging activity. More amounts of Fe and Zn found in Amanita caesaria as compared to A. loosii. The examined mushroom species were rich in protein and carbohydrate. The variations in both edible mushrooms are attributed to different species and/or growth stages. Results obtained through various nutritional and biochemical analysis b exhibited the exploitable potential of these species for cultivation and commercialization. KEY WORDS: Distribution, Antioxidants, Amanita. INTRODUCTION Mushrooms are widely distributed all over the mushroom species collected from different parts of world world and some of them have nutraceutical and have been reported for usefulness of their antioxidant pharmaceutical importance. In general the fruiting bodies of activity [9]. Still there are several varieties of wild edible mushrooms contain about 56.8% carbohydrate, 25.05% mushrooms whose nutritive and medicinal properties have protein, 5. 7 % fat and 12. 5 % ash on a dry weight basis [1]. not been studied. Amanita caeserea and A. loosii from Mushrooms are also gaining importance due to high protein Odisha, India are one such edible mushroom is required to content (19-35 %) in comparison to wheat (13.3%) and to be studied in detail. milk (25.2 %); besides low fat content and vitamin C and B The Genus Amanita contains about 600 species [2-4]. In addition the mushrooms are important source of including some of most toxic as well as some edible species. food and gain income in both developing and developed In India approximately 100 species are reported [10]. It is countries [5,6]. found in North Africa and southern Europe, particularly in Edible mushrooms attract much attention due to its the hills of northern Italy. The mushroom is also distributed medicinal value. Mushrooms are reported to protect heart in Hungary, Mexico and China. Amanita caesarea is listed health, prevent the risk of cancer and help to control blood in the Red Data book of Ukraine and it is protected by law sugar level etc [7]. Many of the diseases/disorders are in Croatia, Slovenia. This species has also been reported in mainly linked to oxidative stress due to free radical Arunachal Pradesh and Khasi hill of Meghalaya in India. formation Barros et al [8] reported the presence of The species has been found in Similipal forest of Odisha antioxidant compounds in food diets, which act as agents to India. It has been observed that local tribals use it is as one reduce oxidative damage in human body. Several of their food items. Corresponding Author:- Nibha Gupta Email:- [email protected] 125 Nibha Gupta. et al. / Vol 4 / Issue 3 / 2014 / 124-129. Similipal, located in the northern Odisha has a (Analytic Jena, Germany). D-glucose at concentration of green moist deciduous forests and climate in spring and 0.5mg/ml used to make standard. The mean value expressed autumn provides ideal conditions for fungal growth with in mg of carbohydrate per gram of mushroom sample. temperature ranging between 18-28°C. Local tribal community utilizes the forest product like mushrooms Estimation of reducing sugar (wild) for their livelihood by collecting and selling them in Reducing sugar was determined by treating 100 mg local tribal market with minimal cost. Only these people dried powder with 5 ml of 80% ethanol [13]. The reaction know edible mushrooms and are able to identify them. mixture was heated at 90 °C till the complete evaporation of Tribal forests dwellers are finding few wild edible ethanol. Residue was added with 5 ml of ethanol and mixed. mushrooms and use them as food. Due to scattered and 100 µl of this was added with 3 ml of DNS reagent (200mg minimal population of some wild mushrooms in the area, crystalline phenol + 50 mg sodium sulphite in 100 ml of 1% they may not include as components of their livelihood. NaOH).The reaction mixture was kept in boiling water bath The two species covered in this paper i. e. Amanita for 5 min. followed by addition of 1ml 40% Rochelle’s salt caesaria and A. loosii have not been explored till date for (Sod. Potassium tartarate).The absorbance was measured at their taxonomic characterization, identification and 510 nm. Glucose was used as standard to calculate the specialty for nutritional and medicinal properties though concentration of reducing sugar and expressed in terms of local people use it occasionally as food. Hence, it is mg per gram dry weight of the samples. necessary to have some basic information to avoid the The amount of non reducing sugar was calculated poisoning factors associated with some the mushrooms. as differential amount of total carbohydrate and reducing Hence, the samples collected from this area were evaluated sugar and expressed in terms of g/100g of dry wt. of for some nutritional and antioxidant properties and samples. described here. DPPH Free Radical scavenging activity MATERIALS & METHODS The DPPH activity was estimated in the Collection and characterization of Mushrooms methanolic extracts by a colorimetric method [14]. 1 gm of Mushrooms samples: Healthy, fresh and succulent samples was added with 20 ml methanol. Methanolic extract mushroom from two species of Amanita caesarea and A. was added with 2 ml of DPPH solution (1:2) and incubated loosii were collected from Tropical moist deciduous and for 30 min. in dark after vigorous mixing. Absorbance was semi evergreen forest of Similipal. measured at 517 .Scavenging activity of each extract on Macroscopic examination for the pileus, stipe, veil, DPPH radical was calculated using the following equation: ring, volva, lamellae and gills etc. for the identification was Scavenging activity (%) = (1- Absorbance of done according to Largent, 1981).The mushrooms samples sample/Absorbance of control) ×100. were carried into the laboratory and preserved in AEAC: (Ascorbic acid Equivalent Antioxidant Capacity) Microbiology laboratory of Regional Plant Resource Centre, was calculated by calibrating the value of above absorbance Bhubaneswar, Odisha, India as specimen as well as further in standard ascorbic acid curve and expressed in mg per analytical and biochemical processing. gram of dried sample. Estimation of protein Determination of Carotenoid content in mushroom The protein content was determined from dried The carotenoid was estimated in 500 mg of dried powder of mushroom (100 mesh size). 1 gram of dried mushroom powder treated with 10 ml of 80% acetone and powder of mushroom was treated with 10 ml of phosphate centrifuged at 3000 rpm for 10 minutes at 4⁰ C. This buffer (pH 7.6), and centrifuged fro 20 min at 8000rpm at procedure was repeated until the residue became colorless. 20 °C. 100 µl of supernatant was mixed with 5 ml of The residue was made upto 10ml with 80% acetone and Bradford, incubated in dark (10 min.) and absorbance was measured for absorbance at 480, 645 and 663 nm recorded at 595nm. The values were expressed in mg separately. The quantity of carotenoids was calculated by protein per gram of dried sample [11]. using following formula and values were expressed in mg/gm by using formula [15] Estimation of Total carbohydrate Carotenoid = A.480+ (0.114×A.663 - 0.638 × A.645). The total carbohydrate was estimated by Phenol Where A= Absorbance. Sulphuric acid method [12]. The total carbohydrate content in dried mushroom powder was estimated by boiling of 100 Estimation of Flavonoids mg samples in 5 ml of 2.5 N HCl. The flavonoid content of dried sample was The mixture was neutralized with sodium estimated from methanolic extract. 100 µl of extract was carbonate until the effervescence ceases. The final volume diluted with 1.5 ml of methanol and incubated for 5 min at was made up to 100 ml and centrifuged. The absorbance room temperature. 0.1ml of AlCl3 was added and again was measured at 490 nm using UV-spectrophotometer incubated at room temperature for 5 min. The reaction 126 Nibha Gupta. et al. / Vol 4 / Issue 3 / 2014 / 124-129. mixture was mixed with 0.1ml of Potassium acetate (1M) (g) Estimation of mineral content and total volume was made upto 5 ml with dist. water. All the samples were weighed 0.5gram and kept in 100µl of methanolic extract was taken in a clean 10 ml concentrated Nitric acid overnight. Then the samples test tube later 1.5ml of methanol was added and incubated were heated till the emergence of white fume and treated for 5 minutes at room temperature then 0.1 ml of AlCl3 was with diacid (Hydrochloric acid and Perchloric acid in 2:1) added again followed by 5 minutes of incubation at room with the help of Microwave digestion system (Milestone temperature followed by addition of Potassium acetate, total stat D, Germany) .The sample volume made up to 100 ml volume was made upto 5ml, finally absorbance was with distilled water.
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