191 Journal homepage: www.fia.usv.ro/fiajournal Journal of Faculty of Food Engineering, Ştefan cel Mare University of Suceava, Romania Volume XVIII, Issue 3- 2019, pag.191 - 200 EFFECT OF RIPENING STAGE ON ORGANIC ACID PROFILES AND ANTINUTRIENT CONTENTS OF THREE SPECIES OF WILD EDIBLE MUSHROOM RUSSULA SSP. Jaures Oscar GBOTOGNON 1 , *Michel Djary KOFFI 2 , Hubert Kouassi KONAN 1 and Eugène Jean Parfait KOUADIO 1 1Laboratory of Biochemistry and Food Technology, Nangui Abrogoua University, 02 BP 801, Abidjan 02, Côte d’Ivoire. 2Laboratory of Biotechnology, Felix Houphouet Boigny University, 22 BP 582 Abidjan 22, Côte d’Ivoire. djaryss@yahoo.fr *Corresponding author Received 1st September 2019, accepted 28th September 2019 Abstract: The present study was aimed at investigating the antinutrient and organic acid contents of three wild edible mushrooms from Russula genus as a function of their ripening stage. Fresh mushrooms Russula lepida, Russula mustelina and Russula delica were harvested from their natural habit in Brobo area’s (7°43'0" N and 4°42'0" W) in center Côte d’Ivoire. The fresh mushrooms were oven dried and ground to obtain the crude flour. Phytate and oxalate contents were investigated using standard colorimetric methods, while organic acid profiles were performed by using HPLC analytical methods. The antinutrient composition showed that the greatest content in phytates (1.60±0.10 mg/100 g) and oxalates (5.17±0.01 mg/ 100 g) were observed respectively, in R. lepida and R. mustelina immature fruiting bodies. These values are much lower than the standard safe limits. As regards organic acid profiles, they revealed the presence of at least fourteen organic acids namely benzoic, oxalic, fumaric, succinic, malic, tartaric, ascorbic, citric, lactic, adipic, propionic, formic, shikimic and acetic acids. The main organic acids in R. lepida were lactic (49.70 mg/ 100 g DW, for post- mature stage), fumaric (36.00 and 31.22 mg/ 100 g DW, respectively for immature and mature stage), citric (21.60 and 31.60 mg/ 100 g DW, respectively for immature and mature stage) and succinic (21.00 mg/ 100 g DW for post-mature stage) acids. Lactic acid (47.90 mg/ 100 g DW) was the dominating organic acid in R. mustelina at immature stage, whereas citric (30.20 mg/ 100 g DW at post-mature stage) and ascorbic acids (21.80 mg/ 100 g DW at post-mature stage) were major in R. delica. R. lepida showed the higher total organic acid contents (ranging from 113.13 mg/ 100 g in immature stage to 103.31 mg/ 100 g in post-mature stage) whatever the ripening stage. As for R. mustelina, the immature stage was better (100.02 mg/ 100 g DW), while R. delica showed better total organic acid content (83.33 mg/ 100 g DW) at post-mature stage. Keywords: Organic acids, edible mushroom, ripening stage, Russula lepida, Russula mustelina, Russula delica. 1. Introduction Mushrooms are the best-known example of macrofungi which are wild edible fungi species with large and visible fruiting bodies [1]. Mushrooms species having a close relationship with the host (usually a tree) are mycorrhizal (symbiotic). Those living on dead organic matter are saprotrophic (saprophytes) while species that live on other species in a non- symbiotic relationship are parasitic species http://www.fia.usv.ro/fiajournal mailto:djaryss@yahoo.fr Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 192 [2]. The mycorrhizal species play a vital ecological role through the symbiotic relationships that they form with trees. They enable trees to grow in nutrient-poor soils. For example, the trees of the miombo woodland of central and southern Africa and the woodland itself would not exist without their fungal partners [3, 1]. Moreover, mushrooms are excellent bio- degraders since they decompose organic waste which releases the nutrients in celluloses, hemicelluloses and lignin. They therefore help in cleaning the environment and recycling the nutrients [4]. The increased interest in consumption of mushrooms as food is as a result of their nutritional, antioxidant and therapeutic values [5]. According to Boa [1], about 1200 species of fungi are used in eighty- five different countries for their gastronomic value and/or medicinal properties. Indeed, wild edible mushrooms are found to be highly rich in proteins, minerals, vitamins, crude fibre and carbohydrate with low fat and oil content [6, 7, 8]. Dué et al. [8] reported that they could be used as valuable substitutes for meat or fish especially in developing countries for malnourished children suffering from kwashiorkor (a protein deficiency condition) and for pregnant women. Chittaragi and Naika [9] argue that mushrooms can provide balancing diet compounds in enough quantities for human nutrition. Due to the low fat and oil content, they are recommended as good source of food supplement for patients with cardiac problems or at risk with lipid induced disorders. Also, several mushroom species have been described to exhibit varied biological and medicinal properties such as antioxydants, antibacterial, antifungal, antiviral, anti-inflammatory, antimutagenic, antitumoral and anti- diabetic activities [10, 11, 12, 13, 14, 15]. Therefore, mushrooms have been categorized as therapeutic foods [16]. The bioactive and taste properties of mushrooms are conferred by compounds such as phenolic derivatives and organic acids [17]. Phenolic compounds and organic acids (especially malic, tartaric, citric, and succinic acids) may have a protective role against various diseases due to their antioxidant activity [18, 19]. Furthermore, organic acids were reported to be responsible of organoleptic characteristics of fruits and vegetables, especially mushrooms [20]. In the center of Côte d’Ivoire, wild edible ectomycorrhizal mushrooms of Russula genus are widely collected and consumed in many households or sold for the improvement of icomes and livelihood of farmers [21]. The species Russula lepida, Russula mustelina and Russula delica were previously investigated for their bioactive compounds and antioxidative properties [22]. Considering the importance of organic acids, the present paper focuses on their acid organic profile and antinutrient contents as a function of their ripening stage. 2. Materials and methods Mushroom collection and preparation of sample Fresh mushrooms were harvested at different ripening stages (immature, mature and post-mature) from their natural habit in Brobo area’s (7°43'0" N and 4°42'0" W) in Côte d’Ivoire. Taxonomic identification was achieved by Dr Souleymane Yorou Nourou (Abome Calavy University of Benin/ Munich University of Germany), as R. lepida, R. mustelina and R. delica. After collection, mushrooms were placed in a cooler with ice to keep their freshness and then Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 193 transported to the laboratory for flour preparation. Fresh mushrooms (500 g) were cleaned to distilled water then, sorted and free from any kind of waste, drained, and dried at 45°C in an oven for 48 h. Dried mushrooms were ground using a blender to obtain the crude flours. Antinutritional factors analysis The titration method as described by Day and Underwood [23] was followed. 1 g of sample was weighed into 100 mL conical flask. 75 mL 3 MH2SO4 was added and stirred for 1 h with a magnetic stirrer. This was filtered using a Whatman No 1 filter paper. 25 mL of the filtrate was then taken and titrated while hot against 0.05 M KMnO4 solution until a faint pink colour persisted for at least 30 s. The oxalate content was then calculated by taking 1 mL of 0.05 m KMnO4 as equivalent to 2.2 mg oxalate [24]. Phytate was extracted according to the procedure described by Mohammed et al. [25]. 1.0 g Sample was extracted with 3% tri-chloro acetic acid (TCA) at 37°C for 45 min. with simple shaking followed by centrifugation and extractation by using anion exchange column. The extracted phytate (0.2 mL) was mixed with 4.6 ml of distilled water and 0.2 mL of chromogenic solution and the tubes were heated in a water bath at 95°C for 30 min, and then were allowed to cool. The developed color was read at 830 nm against blank. Standard phytate solution was prepared by dissolving sodium phytate in distilled water to prepare different phytate concentrations as described above in the tested samples. The amount of phytate in the tested samples was expressed as mg phytate/100 g sample. Extraction of Organic Acids The organic acids of each dried sample of mushroom were extracted according Hasib et al. [26] method by grinding (Waring Blendor, Polychimie, Abidjan, Côte d’Ivoire) in distilled water (1:10, w/v) and clarified by centrifuging at 4000 rpm for 30 minutes. The supernatant was first filtered through Whatmann 4 paper, then through 0.45 μm filter (Millipore; Sartorius AG, Goëttingen, Germany) and stored at −20˚C prior further use. HPLC Analysis of Organic Acids The separation of the organic acids was carried out as previously reported [22] by using a system consisting of an analytical HPLC unit (Shimadzu Corporation, Japan) in conjunction with a column heating device set at 35˚C with the aid of an oven Meta Therm TM (Interchrom, France), with an ions exclusion column ICSep ICE ORH-801 (40 cm × 5 μm, In terchom, France). The system was also coupled to a pump (Shimadzu LC-6A Liquid Chromatograph), a UV detector (Shimadzu SPD-6A UV Spectrophotometric Detector) and an integrator (Shimadzu Chromatopac CR 6A). Elution was carried out isocratically with sulphuric acid 0.04 N, at a solvent flow rate of de 0.6 mL/min and detection was performed at 210 nm. Organic acids in mushroom extracts were identified by comparing the retention times and spectral data obtained from standards under the same conditions. Quantitation was performed by comparing the peak areas with those of the respective external standards. Statistical analysis All chemical analyses and assays were performed in triplicate, unless otherwise indicated. Results were expressed as mean values ± standard deviation (SD). Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 194 Analysis of variance (ANOVA) followed by Duncan’s test was performed to test for differences between means by employing XLstat 2019 statistical software. Significance of differences was defined at the 5% level (P<0.05). Principal component analysis (PCA) was also used in order to discover relationships between independent variables (mushroom species and organic acids). 3. Results and discussion Antinutritional factors The phytate and oxalate contents as a function of ripening stage of the three wild edible mushroom Russula species are presented in table 1. These results showed that the measured antinutritional parameters decreased significantly during ripening of the different mushroom species. We observed the highest content of phytate (1.60±0.10 mg/100 g) in R. lepida, especially in immature fruiting bodies. Gaur et al. [27] reported low values of phytate ranging from 0.11 to 0.19 mg/ 100 g for six selected edible mushrooms in India. However, the phytate contents of the present study were much lower than those found for some cultivated and wild edible mushrooms collected from Ethiopia, ranged from 31.3 to 242.8 mg/ 100 g DW [28]. Furthermore, the phytate contents of studied Russula species are over13 times lower than the standard safe limit of 22.10 mg/ 100 g [29]. This suggest that whatever the ripening stage, the three studied mushroom species could be consumed without risk of toxicity associated with phytate concentration. As regards oxalate, the highest content (5.17±0.01 mg/ 100 g) was estimated in R. mustelina immature fruiting bodies. Oxalate contents obtained in this study were higher than that found for the Oyster mushroom Pleurotus ostreatus (0.41 mg/ 100 g) [30], but quite lower than the tolerable limit of 105 mg/100 g recommended by World Health Organization. This suggests the safe for consumption of these mushrooms at any ripening stage. Moreover, it’s noteworthy that food processes such as boiling or cooking, fermentation and milling could reduce drastically or remove antinutrient elements [31]. Table 1. Antinutritional factors content as a function of ripening stage of three Russula species Parameters Stage Species R. lepida R. mustelina R. delica Phytate (mg/100g) I 0.60±0.20ᵃA 1.07±0.02ᵃB 1.60±0.10ᵇC M 0.40±0.30ᵃA 0.56±0.22ᵇA 1.46±0.25ᵃᵇB PM 0.23±0.23ᵃA 0.53±0.15ᵇA 1.20±0.10ᵃB Oxalate (mg/100g) I 2.83±0.02ᶜA 5.17±0.01ᶜC 4.34±0.03ᶜB M 2.10±0.01ᵇA 4.13±0.02ᵇC 3.26±0.02ᵇB PM 0.77±0.02ᵃA 3.03±0.01ᵃB 3.08±0.01ᵃC Values are mean ± standard deviation of three measurements (n = 3). I: immature; M: mature; PM: post-mature. a,b,cIdentical script indicate no significant difference between mean values in line. A,B,CIdentical script indicate no significant difference between mean values in column. Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 195 Organic acids composition Organic acids are categorized as important food components for the formation of taste and flavour, and determination of the quality and safety of food and food products [32]. They generate important effects on food such as sensorial, antioxidant and acidifying properties [33]. The HPLC-UV analysis (figure not shown) allowed the identification of at least fourteen organic acids namely benzoic, oxalic, fumaric, succinic, malic, tartaric, ascorbic, citric, lactic, adipic, propionic, formic, shikimic and acetic acids. Most of these organic acids has already been found in edible mushrooms [34, 35]. Results summarized in table 2 indicated that the three mushroom species were characterized by a high variation in their organic acid content. Moreover, these organic acid contents were strictly dependent on ripening stage. For the three ripening stages, R. lepida showed the highest total organic acid contents (ranging from 103.31 to 113.13 mg/ 100 g DW), mainly consisting of lactic (49.70 mg/ 100 g DW, for post-mature stage), fumaric (36.00 and 31.22 mg/ 100 g DW, respectively for immature and mature stage), citric (21.60 and 31.60 mg/ 100 g DW, respectively for immature and mature stage) and succinic (21.00 mg/ 100 g DW for post-mature stage) acids. Lactic acid is the product of glycolysis under anaerobic condition. In food, it’s characterized as a natural flavour enhancer, mild and lingering [36]. Fumaric and citric acids are well known for their antimicrobial and antioxidant properties making them important agents in the prevention of mushroom browning [37]. R. mustelina presented total acid content of 100.02 mg/ 100 g DW at immature stage, with dominating lactic acid (47.90 mg/ 100 g DW), whereas R. delica had a total acid content of 83.33 mg/ 100 g DW at post- mature stage. The major acids in R. delica were citric acid (30.20 mg/ 100 g DW at post-mature stage) followed by ascorbic acid (21.80 mg/ 100 g DW at post-mature stage). Total organic acid contents found in this study were higher than those found for Fustulina hepatica [38]. Table 2. Organic acids content as a function of ripening stage of three Russula species Organic Acids Retention time (min) Stage Species R. lepida R. mustelina R. delica Butyric acid 2.53 I nd nd 0.06±0.01aA M nd 0.05±0.01aA nd PM nd nd nd Benzoic acid 5.62 I 4.00±0.02bB nd 0.11±0.01aA M 3.63±0.03aA nd nd PM nd 1.26±0.02aA 3.84±0.01bB Oxalic acid 9.66 I 0.62±0.03bC 0.04±0.01aA 0.61±0.01bA M 0.52±0.03bB 0.41±0.01aC 0.61±0.01cA PM 0.06±0.01aA 0.21±0.02bB 1.14±0.01cB Fumaric acid 10.48 I 36.00±0.04bB 2.62±0.02aB nd M 31.20±0.02cA 2.14±0.01aA 6.47±0.02bB PM nd 3.10±0.02aC 3.14±0.01aA Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 196 Succinic acid 11.25 I 5.31±0.02bB 17.70±0.03aC nd M 4.58±0.02bA 0.76±0.02aA nd PM 21.00±0.03cC 1.88±0.3aB 8.16±0.01bA Malic acid 12.95 I 3.27±0.01bB nd 0.50±0.01aA M 0.43±0.02aA 2.49±0.02bA 5.19±0.06cB PM nd 2.85±0.02aB 9.82±0.03bC Tartaric acid 15.54 I 12.10±0.02bB 10.10±0.03cC 3.76±0.02aA M 11.40±0.03bB 1.29±0.02aA 6.96±0.01bB PM 9.27±0.02bA 9.45±0.01bB 13.10±0.01aC Ascorbic acid 16.31 I 7.87±0.02bB 3.48±0.02aB nd M 8.43±0.02bC 0.48±0.35aA nd PM 4.33±0.02aA nd 21.80±0.05bA Citric acid 17.7 I 29.60±0.03bB 5.90±0.01aC 30.20±0.01bC M 31.60±0.02cC 1.50±0.02aA 2.46±0.03bB PM 7.94±0.02cA 2.90±0.00bB 2.16±0.02aA Lactic acid 20.75 I 6.03±0.03bA 47.90±0.01cC 0.46±0.02aA M 5.98±0.02bA 5.01±0.01aA 11.40±0.01cC PM 49.70±0.02cB 12.10±0.01bB 7.52±0.03aB Adipic acid 22.42 I 0.79±0.02cC 0.52±0.03bA 0.02±0.00aA M 0.54±0.02aA 1.13±0.02bB 2.22±0.02cB PM 0.60±0.01aB 1.10±0.01bB nd Propionic acid 24.15 I 3.39±0.02cB 1.53±0.03bA 0.94±0.02aB M 4.14±0.02cC 0.41±0.01aB 0.63±0.02bA PM 2.00±0.02bA 0.29±0.01aC 5.45±0.02cC Formic acid 28.04 I 1.42±0.03bC 0.24±0.02aA 2.52±0.02cC M 0.99±0.02cB 0.21±0.01aA 0.59±0.02bB PM 0.26±0.02aA 0.51±0.02bB 0.52±0.02bA Shikimic acid 34.22 I 2.73±0.02bA 7.24±0.02cB 0.02±0.00aA M 3.78±0.01cB 1.59±0.01aA 1.65±0.02bB PM 5.60±0.02bC 1.85±0.01aA 6.68±0.02cC Acetic acid 36.46 I nd 2.75±0.01aC nd M nd 0.49±0.01aA 1.43±0.04bA PM 2.55±0.02bA 0.88±0.02aB nd Total I 113.13±0.03 100.02±0.02 39.2±0.02 M 107.22±0.02 17.96±0.01 39.61±0.02 PM 103.31±0.02 38.38±0.03 83.33±0.02 Values are mean ± standard deviation of three measurements (n = 3). I: immature; M: mature; PM: post-mature. a,b,cIdentical script indicate no significant difference between mean values in line. A,B,CIdentical script indicate no significant difference between mean values in column. Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 197 The principal component analysis (PCA) based on the organic acid contents obtained for the three mushroom species as a function of ripening stage is presented in Figure 1. In terms of information, the principal components 1 and 2 contained more than 80 % (81.62) of the total variance explained with eigenvalues for the first two components greater than unity (respectively, 4.226 and 2.922) (Table 3). This value of the total variability is relatively significant. Also, the Kaiser- Meyer-Olkin (KMO) criterion is 0.761 (Table 4) suggesting that sampling is acceptable for the validity of PCA test in this study. Table 3. Eigen values of the main components F1 F2 Eigenvalues 4.226 2.922 Variability (%) 52.262 29.220 Cumulated Percentage 52.262 81.482 Table 4. Sampling accuracy RT 0.149 L1 0.770 L2 0.791 L3 0.802 M1 0.657 M2 0.656 M3 0.957 D1 0.855 D2 0.715 D3 0.769 KMO 0.761 The PCA analysis (Figure 1) makes it possible to distinguish the mushroom species in two large groups. One formed by R. delica and R. lepida and the other represented by R. mustelina. The first one better expresses the axis of the first component (52.41% of the total variance explained) and the second the axis of the second component (29.21% of the total variance explained). The first group is characterized by the relationship R. delica and R. lepida - fumaric, citric, shikimic, malic, tartaric, ascorbic, succinic acids. For R. mustelina (which constitutes the second group), PCA analysis revealed a close relation with lactic, succinic and tartaric acids. The ripening stages were used as additional variables for the PCA analysis (Figure 1). Results suggest a difference in the influence of the ripening stages on the organic acid contents, contributing in different ways to the constitution the study axes. Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 198 Fig. 1. PCA analysis for organic acids content as a function of ripening stage of three Russula species 4. Conclusion Based on the present study, it appears that Russula species from Côte d’Ivoire contained a wide range of organic acids (fourteen). These organic acids are unevenly distributed (with widely varying proportions) according to the species and the ripening stages. R. lepida shows the higher total organic acid contents for all ripening stages mainly consisting of lactic, fumaric, citric and succinic. For R. delica, the major organic acids were citric and ascorbic acids found at post-mature stage, while lactic acid was the dominating acid at the immature stage in R. mustelina. The PCA analysis suggests a relationship between R. delica and R. lepida which would forme a group, whereas R. mustelina constitutes another group. 5. References [1]. BOA E., Wild Edible Fungi.A Global Overview of Their Use and Importance to People.Non-wood Forest Products Series no. 17. Rome: FAO, (2004). [2]. KALAC P., Chemical composition and nutritional value of European species of wild growing mushrooms Nova Science Publishers, (2012). [3]. CAMPBELL B., ed. 1996. The miombo in transition: woodlands and welfare in Africa. Bogor, Indonesia, Centre for International Forestry Research. 266 pp, (1996). [4]. OEI P., Mushroom cultivation: appropriate technology for mushroom growers. Mushroom Cultivation, 3rd edition, Leiden, The Netherlands. pp.xii + 429 pp, (2003). [5]. EGWIM E.C., ELEM R.C., EGWUCHE R.U., Proximate composition, phytochemical screening and antioxidant activity of ten selected wild edible Nigerian mushrooms. American Journal of Food and Nutrition, 1(2): 89-94, (2011). [6]. OKWULEHIE I.C., ODUNZE E.I., Evaluation of the myco-chemical and mineral composition of some tropical edible Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 199 mushrooms. Journal of Sustainable Agriculture and Environment, 6: 163 – 170, (2004). [7]. DUE E.A., KOFFI D.M., DIGBEU Y.D., Biochemical and functional properties of two wild edible mushrooms (Volvariella volvacea and Armillaria mellea) consumed as protein substitutes in South Côte d’Ivoire. Journal of Chemical, Biological and Physical Sciences, 6(4): 1197 – 1206, (2016) [8]. DUE E.A., KOFFI D.M., DIGBEU Y.D., Physicochemical and Functional Properties of Flour from the Wild Edible Mushroom Termitomyces heimii Natarajan Harvested in Côte d’Ivoire. Turkish Journal of Agriculture - Food Science and Technology, 4(8): 651 – 655, (2016). [9]. CHITTARAGI A., NAIKA R., Physico chemical and Primary Biochemical Studies of Hygrocybe cantharellus Collected from Western Ghats region of Haniya, Shimoga (Dist) Karnataka. International Journal of Advances in Pharmacy, Biology and Chemistry, 3(3): 523 – 528, (2014). [10]. GARCÍA-LAFUENTE A., MORO C., VILLARES A., GUILLAMÓN E., ROSTAGNO M.A., D’ARRIGO M., MARTÍNEZ J.A., Mushrooms as a source of anti-inflammatory agents. American Journal of Community Psychology, 48: 125–141, (2011). [11]. SCHILLACI D., ARIZZA V., GARGANO M.L., VENTURELLA G., Antibacterial activity of Mediterranean Oyster mushrooms, species of genus Pleurotus (higher Basidiomycetes). International Journal of Medicinal Mushrooms, 15(6), 591 – 594, (2013). [12]. PHAN C., LEE G., MACREADIE I., MALEK S., PAMELA D., SABARATNAM, V., Lipid constituents of the edible mushroom, Pleurotus giganteus demonstrate anti-candida activity. Natural Product Communications, 8(12): 1763–1765, (2013). [13]. MA K., HAN J., BAO L., WEI T., LIU H. Two sarcoviolins with antioxidative and α‐glucosidase inhibitory activity from the edible mushroom Sarcodon leucopus collected in Tibet. Journal of Natural Products, 77(4): 942−947, (2014). [14]. AMIRULLAH N.A., ABIDIN N.Z., ABDULLAH N., The potential applications of mushrooms against some facets of atherosclerosis: a review. Food Research International, 105: 517–536, (2018). [15]. MUSZYŃSKA B., GRZYWACZ- KISIELEWSKA A., KAŁA K., GDULA- ARGASIŃSKA J., Anti-inflammatory properties of edible mushrooms: a review. Food Chemistry, 243, 373–381, (2018). [16]. APETORGBOR M.M., APETORGBOR A.K., OBODAI M., Indigenous knowledge and utilization of edible mushrooms in parts of Southern Ghana. Ghana Journal of Forestry, 19 & 20: 19 – 34, (2006). [17]. VALVERDE M.E., HERNÁNDEZ-PÉREZ T., PAREDES-LÓPEZ O., Edible mushrooms: improving human health and promoting quality life. International Journal of Microbiology, 2015: 376 – 387, (2015). [18]. SILVA B.M., ANDRADE P.B., VALENTÃO P., FERRERES F., SEABRA R.M., FERREIRA M.A., Quince (Cydonia oblonga Miller) fruit (pulp, peel, and seed) and jam: antioxidant activity. Journal of Agricultural and Food Chemistry, 52: 4705 – 4712, (2004). [19]. BARROS L., PEREIRA C., FERREIRA I.C.F.R., Optimized analysis of organic acids in edible mushrooms from Portugal by ultra- fast liquid chromatography and photodiode array detection. Food Analytical Methods, 6(1): 309–316, (2013). [20]. VAUGHAN J.G., GEISSLER C.A., In The New Oxford Book of Food Plants; Oxford University Press: New York, p 196, (1997). [21]. KONÉ N.A., YÉO K., KONATÉ K.S., LINSENMAIR E., Socio-economical aspects of the exploitation of Termitomyces fruit bodies in central and southern Côte d’Ivoire: Raising awareness for their sustainable use. Journal of Applied Biosciences, 70: 5580 – 5590, (2013). [22]. KOUASSI K.A., KOUADIO E.J.P, DJÈ M., DUÉ A.E., KOUAMÉ L.P., Edible Ectomycorrhizal Mushrooms Russula spp. of Côte d’Ivoire: Total Phenolic Content, HPLC-Profiles of Phenolic Compounds and Organic Acids, Antioxidant Activities. Journal of Agricultural Chemistry and Environment, 5: 73 – 84, (2016). [23]. DAY R.A., UNDERWOOD A.L., Quantitive analysis 5th ed. Prentice. Hall Publication, p. 701, (1986). [24]. CHINMA C.E., IGYOR M.A., Micronutrients and anti-nutritional contents of selected tropical vegetables grown in Southeast, Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XVIII, Issue 3 – 2019 Jaures Oscar GBOTOGNON, Michel Djary KOFFI, Hubert Kouassi KONAN, Eugène Jean Parfait KOUADIO, Effect of ripening stage on organic acid profiles and antinutrient contents of three species of the wild edible mushroom Russula Ssp., Food and Environment Safety, Volume XVIII, Issue 3 – 2019, pag. 191 - 200 200 Nigeria. Nigerian Food Journal, 25(1): 111– 116, (2007). [25]. MOHAMED A., PERERA P.J., HAFEZ Y.S., New chromophore for phytic acid determination. Cereal Chemistry,63(6):475– 478, (1986). [26]. HASIB A., JAOUAD A., MAHROUZ M., KHOUILI M., HPLC Determination of Organic Acids in Moroccan Apricot. Ciencia y Tecnología Alimentaria, 3, 207 – 211, (2002). [27]. GAUR T., RAO P.B., KUSHWAHA K.P.S., Nutritional and antinutritional components of some selected edible mushroom species. Indian Journal of Natural Products and Resources, 7(2): 155 – 161, (2016). [28]. WOLDEGIORGIS A.Z., ABATE D., HAKI G.D., ZIEGLER G.R., Major, Minor and Toxic Minerals and Anti-Nutrients Composition in Edible Mushrooms Collected from Ethiopia. Journal of Food Processing and Technology, 6: 430. doi:10.4172/2157- 7110.1000430, (2015). [29]. GOUD M.J.P., SURYAM A., LAKSHMIPATHI A.V., CHARYA S.M.A., Extracellular hydrolytic enzyme profiles of certain South Indian basidiomycetes. African Journal of Biotechnology, 8 (3), 354 – 360, (2009). [30]. DURU M., EBOAGWU I., KALU W., ODIKA P. Nutritional, Anti-nutritional and Biochemical Studies on the Oyster Mushroom, Pleurotus ostreatus. EC Nutrition 14.1: 36 – 59, (2019). [31]. SANDBERG A.S., ANDERINE R., HPLC method for determination of inositol tri, tetra, penta- and hexaphosphates in food and intestinal contents. Journal of Food Science, 51: 547 – 50, (1986). [32]. YÜKSEL A.K., YÜKSEL M., ŞAT İ.G., Determination of certain physicochemical characteristics and sensory properties of green tea powder (matcha) added ice creams and detection of their organic acid and mineral contents. GIDA 42(2): 116 – 126, doi: 10.15237/gida.GD16072, (2017). [33]. TORMO M., IZCO J.M., Alternative reversed phase high performance liquid chromatography method to analyse organic acids in dairy products. Journal of Chromatography A, 1033: 305 – 310, (2004). [34]. GĄSECKA M., MAGDZIAK Z., SIWULSKI M., MLECZEK M., Profile of phenolic and organic acids, antioxidant properties and ergosterol content in cultivated and wild growing species of Agaricus. European Food Research and Technology, DOI 10.1007/s00217-017-2952-9, (2017). [35]. MAGDZIAK Z., SIWULSKI M, MLECZEK M., Characteristics of organic acid profiles in 16 species of wild growing edible mushrooms, Journal of Environmental Science and Health, Part B, 52: 10, 784 – 789, (2017), DOI: 10.1080/03601234.2017.1356676. [36]. JARRETT T.N., Acids in Confections. Food- grade organic acids in confections offer ranges in sourness, intensity and linger; balance sweetness; and round out the taste profile; and perform other functions. The Manufacturing Confectioner, pp 58-63, (2012). [37]. RIBEIRO B., ANDRADE P.B., BAPTISTA P., BARROS L., FERREIRA I.C.F.R., SEABRA R.M., VALENTÃO P., Leucopaxillus giganteus mycelium: Effect of nitrogen source on organic acids and alkaloids. Journal of Agricultural and Food Chemistry, 56: 4769-4774, (2008). [38]. RIBEIRO B., VALENTÃO P., BAPTISTA P., SEABRA R.M., ANDRADE P.B., Phenolic compounds, organic acids profiles and antioxidative properties of beefsteak fungus (Fistulina hepatica). Food Chemistry and Toxicology, 45: 805–1813, (2007). 1. Introduction 4. Conclusion