INTRODUCTION:

Edible wild fruits have historically been crucial in supplementing people's diets. These non-domesticated fruit plants flourish in the wild, responding to diverse climatic attributes along with biotic and abiotic stimuli.¹ They act as functional foods because they are high in proteins, polyphenols, amino acids, vitamins, fibers, and minerals including sodium, potassium, magnesium, iron, and calcium which can act as dietary supplements and have a variety of medicinal purposes^2-4 and two billion people worldwide consume them for nourishment and used it for income.⁵ However, their popularity has declined as exotic fruits are introduced. These fruits offer nutritional value, minerals, and immunity to diseases, making them popular in Indian folk medicine, such as Ayurveda.⁶ Huge portion of global population today utilizes herbal medicine to assert their health, therefore plant products have become the primary source of novel pharmaceuticals. Changes in conditions, particularly in the post-COVID age, have prompted the resurgence of traditional health practices as a crucial part of people's reflections of their local cultures. In recent years, there has been a surge in the use of plant-based natural goods, driving up market demand. Specifically, in the post-COVID age, dietary patterns have changed in terms of the consumption of local fruits and vegetables in order to enhance immunity.^7,8Numerous phytochemicals that have a positive impact on health, including antioxidant activity, are produced by plants. These chemicals decrease the production of reactive oxygen species (ROS), which prevents cell destruction and delays or prevents the oxidation of oxidizable molecules. While non-enzymatic antioxidants from dietary sources operate as free radical scavengers, enzyme-based antioxidants, such as SOD, Catalase, and peroxidase, neutralize damaging free radicals ensuring overall health.^3,9,10 Isolating and describing naturally occurring antioxidants from cereals, fruits, vegetables, herbs, spices, and seeds is an objective for researchers, as synthetic antioxidants have low solubility and moderate antioxidant activity.^11-13L-ascorbic acid (L-threo-hex-2-enono-1,4-lactone, ascorbate), also known as Vitamin C, is a crucial vitamin that must be consumed in the diet, cause some animals, including humans, and non-human primates, and some groups of bats and birds, have lost the potency to synthesize ascorbate due to mutations in the coding sequence.^14-16 Ascorbate plays a crucial role in antioxidant and redox reactions, as well as controlling cell differentiation.¹⁷ Wild edible fruits in Odisha are consumed in different ways based on their ripeness, taste, and culinary preferences, some are consumed in their ripe or unripe conditions while others are cooked and some are pickled.¹⁸ It is evident that the tribal populations dwell and reside along the forest and consume the wild edible fruits in their daily diet thus it can be inferred from the anecdotal evidence that they are rich in nutritional and therapeutic values. For instance, the fruits of Artocarpus lakoocha Roxb. known as granthiphala, Kshudra Panasa, Pitanaasha in Ayurved commonly referred to as monkey jackfruit, and Jeuta in Odia (Odisha), are distinctive and edible.¹⁹ The fruits are usually eaten raw or made into curries and chutneys or sun-dried to be used as a replacement for mango and tamarind is commonly used by tribes of Paruja, Gadaba, Bhatra, Saora, Gonda, Kondha, Khola, Santhal, Bathudi, Bhumija, Khadia and Mankirdia belonging to the state of Odisha.²⁰ The fruits of Mimusops elengi Linn., sometimes referred to as Spanish Cherry, and Baula in Odia (Odisha), turn red, orange-yellow, or brown when ripe. The fruits are a rich source of carbohydrates and are generally eaten raw or their dried pulps are infused into tea for their unique flavor and are utilized in the formulation of traditional medicines for the intervention of many chronic diseases consumed by the Kondha, Koya, Paraja, Saura, Langia, Bonda, Paika, Bhumia and Bhatra tribal communities of Odisha.²¹Artocarpus lakoocha Roxb. (Jeuta) Moraceae Substantial deciduous tree acts as a liver tonic, shows antibacterial and antiparasitic qualities, inhibits pancreatic lipase activity, and possesses potential cytotoxicity.^22,23Mimusops elengi Linn. (Baula) Sapotaceae Small to medium-sized, dense tropical evergreen tree applied in managing long-term dysentery, migraines, dental pain, and additional oral ailments^24,25; demonstrates fever-reducing properties, and alleviates constipation and dysentery.²⁶But there is a lack of reports on the potentiality of the fruits and their significance. Traditional knowledge thus provides valuable insights and acts as a guide for scientific investigations; however, empirical evidence is crucial to fully understand and utilize the nutritional and therapeutic potential of these fruits.

MATERIALS AND METHODS:

Samples and Sampling Sites:

The target fruit samples (Artocarpus lakoocha and Mimusops elengi) were collected from two different agro-climatic zones i.e. East and South Eastern Coastal Plain (ESECP) and Mid Central table land (MCTL) of Odisha for the analysis of ascorbic acid along with different antioxidants to know the potentiality of the wild fruit species at their different maturation stages.The collections of fruits were done from the rich floral and faunal diversities of Odisha (Table 2) that belong to the East and South Eastern Coastal Plain and Mid Central Table Land between October 2021-December 2022. Ethno-botanical and socio-economic significance was explored through interaction and discussions with tribals and people living in that concerned geographical area. Fruits were botanically confirmed using the reference book “The Flora of Odisha”²⁷ and compared with authenticated herbarium specimens from the Regional Plant Resource Centre in Bhubaneswar, Odisha.

Sample Processing:

Fruit samples utilized for the validation of the Ascorbic Acid, Phenol, POX, CAT, and SOD were kept at -20^ͦ C until their analysis. For total antioxidant content (DPPH, FRAP assay), small pieces of fruit samples were dried

at 45^ͦ C - 50^ͦ C in Hot air Oven. After drying, fruits were finely powdered and kept in an air-tight jar in dark condition until further use.

Reagents and Chemicals:

Standards (98% >Purity) such as Gallic acid, L-Ascorbic Acid, DPPH (2, 2-diphenyl-1-picrylhydrazyl), and Metaphosphoric Acid were procured from Sigma-Aldrich. Other chemicals used were 2,4-dichlorophenol-indophenol (DCPIP), oxalic acid, acetic acid, thiourea, bromine water, 2,4-dinitrophenylhydrazine (DNPH), Sulphuric acid, O-dianisidine, Hydrogen Peroxide, TPTZ, NBT, Aluminium chloride Hexahydrate (AlCl_3.6H₂O), Folin- Ciocaltteus reagent (FCR), Sulphosalicylic acid, Phosphate buffer, Acetate Buffer, Ferric Chloride, Ninhydrin, Hydrochloric Acid, Sodium carbonate, Methanol and Ethanol.

Moisture analysis:

The moisture content of the samples was evaluated through a moisture analyzer (Aczet Make).

Extraction of the sample for quantification of ascorbic acid through the volumetric and Spectrophotometric method:

The fresh fruit pulp (0.5gm) was homogenized with a mortar and pestle in 5ml of 4% oxalic acid. The extract was filtered and was made up to the mark with the reagent followed by the methods of²⁸ with slight modifications for volumetric method. Further for spectrophotometric analysis, the fresh fruit sample (10mg) was homogenized taking 50ml of 3%Metaphosphoric Acid in 10% glacial acetic acid and mixed gently to get a homogenous dispersion and was diluted by adding the extraction solvent. The solution was Centrifuged (Eppendorf cooling Centrifuge, 5430 R) at 6500rpm for 25min at 4^oC and filtered sample was used for evaluation of Ascorbic Acid using the methods described by²⁹ with modifications.

Extraction of sample for Antioxidant Assay:

For extraction of the fruit sample, 5-7gm of powdered sample was taken out in the extraction thimble. The extraction procedure was followed using the Soxhlet Apparatus (Rivotech Rivera). Methanol and ethanol were used as extraction solvent and the extraction was done for about 36-48hours.³ The extraction procedure was done at about 20^ͦC-30^ͦC temperature.

Quantification of Ascorbic Acid through Volumetric Method:

The prepared sample solution (5ml) was taken out in a conical flask and 10ml of oxalic acid was added to it and titrated against 2, 4-dichlorophenol-indophenol (DCPIP) dye (V₁). The endpoint was the appearance of a pink colour which persists for a few minutes. The volume of dye consumed was noticed and it was considered as the amount of ascorbic acid.²⁸ Ascorbic Acid content was calculated through the standard calibration plot for which the L- Ascorbic Acid (Sigma-Aldrich) was used.

Quantification of Ascorbic Acid through Spectrophotometer:

The filtrate fruit sample (2ml) was taken in a test tube with the addition of a few drops of bromine water until the solution became coloured. Now to remove the excess amount of bromine, a few drops of 10% thiourea were added in order to obtain a clear solution. To the sample solution, 1ml of DNPH solution was added thoroughly and kept at 37^oC for 3 hours. After 3hours, the test tube was cooled in an ice bath for duration of 30minutes and was mixed with 5ml pre-chilled 85% sulphuric acid. Then the OD was measured at 280nm using a spectrophotometer (Analytik Jena, Spekol 2000, Germany).²⁹

Total Antioxidant assays:

DPPH Radical Scavenging Assay:

Following the procedure of,² the free radical scavenging activities of the fruit samples for the radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) were evaluated using a spectrophotometer (Spekol 2000, Analytik Jena) at 517nm. By graphing, the ascorbic acid standard curve was created. From the ascorbic acid versus absorbance standard curve graph, sample concentration was estimated.

Ferric Reducing Antioxidant Potential Assay (FRAP):

To calculate the total antioxidant capacity, the FRAP assay was carried out^3,30 using TPTZ. The OD was taken at 620nm using a UV-Vis spectrophotometer (Spekol 2000, Analytik Jena).²The FRAP values were calculated using the standard curve and represented in “mM ascorbic acid equivalent (AAE)/g dry weight”.

Total Phenol:

Gallic Acid (Sigma-Aldrich) was used as the Standard for Total Phenol quantification followed by the methods of²using Folin- Ciocalteu reagent. This reaction mixture was incubated for 1hour at room temperature in the dark. The absorbance was taken at 760nm through UV-Vis spectrophotometer (Spekol 2000, Analytik Jena).

Catalase (CAT):

The enzyme extract was developed by grinding 0.5gm of the fresh wild edible fruit sample in 5ml of phosphate buffer of pH 7 by centrifugation (Eppendorf cooling Centrifuge, 5430 R) at 4000rpm at 4^oC for 15 minutes. The supernatant was stored at 4^oC for further use. Then the reaction mixture was prepared. The OD was measured immediately at 240nm through UV-Vis spectrophotometer (Spekol 2000, Analytik Jena) as initial reading followed by final reading after 240 seconds.³

Peroxidase (POX):

The peroxidase enzyme assay was estimated following the method of³¹ which was further slightly modified.³ After peroxidation of O-dianisidine, the OD was taken at 530nm using spectrophotometer with 1 minute interval upto 10minutes. The enzyme activities were expressed in terms of “an average increment in absorbance per minute per gram fresh weight (∆ O.D/min/g f.wt.)”.

SOD (Super Oxide Dismutase Assay):

The SOD activity was estimated by³ calculating its inhibitory potency for photochemical reduction of nitro blue tetrazolium (NBT), by measuring the OD at 560nm in spectrophotometer (Spekol 2000, Analytik Jena, Germany). For control, a non-irradiated reaction mixture was served used and identical solutions without illumination as blank.

Statistical Analysis:

All the grouped data were statistically evaluated with GraphPad Prisim software (8.4.2). Data were expressed as mean±SD. and analyzed statistically using Two-way ANOVA. Differences were considered statistically significant at P<0.05

RESULTS:

Epidemiological studies show that regular consumption of natural antioxidants is linked to a lower risk of cancer, cardiovascular disorder, and chronic illnesses like Alzheimer's disease, heart disease, diabetes, hypertension, and stroke.^11,32,33 In the present research work, two wild edible fruits were chosen for their antioxidant enzyme potency, especially Ascorbic Acid which would validate to be prime criteria for “free radical scavenging” potency in human health and nutrition owing to their far-flung consumption and assimilation in different forest zones of Odisha. The study findings about the antioxidant activity by assessing Ascorbic Acid as well as enzymatic (Catalase, Peroxidase, Super Oxide Dismutase) and non-enzymatic (DPPH, FRAP, Total Phenol) antioxidant activities.

Moisture Content:

From the moisture analysis (Table 1) it was found that A. lakoocha has a moisture content (89.25%) which is the highest for the particular fruit among its three developmental stages collected from the ESECP zone at its ripen stage while M. elengi moisture content was highest (79.42%) at its mature stage collected from MCTL. This variation may be due to the average rainfall, soil, and humidity in that particular agro-climatic zone. The moisture and humidity content in ESECP zone is relatively higher as compared to the MCTL zones of Odisha.³⁴

Quantification of ascorbic acid through Volumetric Method:

From the analysis result (Table 2) it was found the highest amount of ascorbic Acid in Artocarpus lakoocha at its semi-mature stage i.e., 198.75 mg/100gm f.wt. Collected from ESECP. There is a great variation in Ascorbic Acid content in both fruits at its three developmental stages as there is instability of ascorbic acid and interferences. For the fruits of M. elengi result shows that the highest amount of ascorbic acid was found at its Semi-Mature stage (153.76 mg/100gm fwt.) for fruits collected from ESECP and lowest at its ripening stage (99.37 mg/100gm fwt.) whereas for the fruits collected from MCTL highest Ascorbic Acid content found at it ripen stage i.e., 112.08mg/100gm fwt (Figure 1).

Figure 1: Quantification of Ascorbic Acid through Volumetric method.

Quantification of Ascorbic Acid through UV-Vis Spectrophotometer:

The “oxidation of ascorbic acid to dehydroascorbic acid and diketogulonic acid, followed by coupling with 2,4-dinitrophenylhydrazine (DNPH) under strictly regulated conditions to provide red-colored osazones”, was the basis for the spectrophotometric quantification of ascorbic acid. We found a higher amount of Ascorbic Acid in fruits of A. lakoocha (Table 2) at its semi-mature stage (132.16mg/100gm fwt.) collected from the ESECP zone while the least was found in fruits of M. elengi (Figure 2)collected from MCTL zone at its Mature stage (46.2 mg/100gm f. wt.). These variations may be because of the moisture content and climatic conditions that affect the Ascorbic Acid content in fruits.³⁵ For both the fruits, in the volumetric method as well as the spectrophotometric method we observed that the Ascorbic acid was decreasing mostly with the maturation of fruits.

Figure 2: Quantification of Ascorbic Acid through UV-Vis Spectrophotometer.

Total Antioxidant Capacity:

The total antioxidant capacity, assessed through both the DPPH and FRAP assays, indicates that the extracts from the fruit pulp exhibit varying abilities to scavenge free radicals using these methods.

DPPH:

The DPPH assay, the indirect method for assessing antioxidant activity, measures the ability to stabilize the free radical 2,2-diphenyl-1-picrylhydrazyl by reacting with hydrogen donors such as phenol. The antioxidant capacity of wild edible fruits, significant in ethno medicine, was determined using the DPPH assay and expressed as “AEAC (Ascorbic Acid Equivalent Antioxidant Capacity)”.

IC50 value in DPPH assay:

The IC50 values for each sample were determined graphically (Figure 3), representing the concentration in micrograms of dry sample per millilitre that hinders the development of DPPH radicals by 50%. Measurements were conducted in five times to ensure accuracy and reliability. Measurements were taken (n=5).

% of Scavenging activity = (Ac-As/Ac) x 100

Where, Ac=OD at control at 517nm, As= OD of sample at 517nm

Our study and analysis revealed that M. elengi exhibits the highest scavenging activity compared to other samples i.e., 89.79% at its semi-mature stage collected from ESECP and lowest (45.12%) at its ripen stage collected from MCTL among both the fruits. While A. lakoocha shows maximum DPPH activity i.e., 87.89% at it ripen stage collected from MCTL zone (Table 2).

Figure 3: IC 50 value for DPPH % of Scavenging Activity.

FRAP:

The FRAP value assesses how well an antioxidant can convert Fe (3+) to Fe (2+). In the FRAP assay, the antioxidant's capacity to transform the Fe3+/ferricyanide complex into the Fe2+/ferrous form serves as a useful indicator for its antioxidant activity. It is evident from the study that the ripened stage of Mimusops elengi collected from ESECP (Figure 4) has the highest total antioxidant activity (FRAP) i.e., 107.5 mM AEAC/g dry Wt. (Table 2).

Figure 4: Ferric Reducing Antioxidant Power Assay (mM AEAC/g dry wt.).

Total Phenol:

The FCR reagent reacts with the phenols thus causing a colour change to dark blue, which was then quatified using a UV-visible spectrophotometer (n=5). The total phenol content in the fruit samples' methanolic extract was expressed in milligrams of gallic acid equivalents per gram of dry weight, utilizing the equation derived from the standard curve: y= 1.6104x+0.0082, R²=0.981. Results of the current study demonstrated that Mimusops elengi at its mature stage collected from MCTL has the highest phenolic content(Figure 5) i.e., 399.14mg/GAE dry Wt. among the two fruits. There is a significant variation found in Phenol content among the two fruits. For the fruit of Artocarpus lakoocha the highest Phenolic content was found at its ripen stage i.e., 289.44mg/GAE dry wt. which is collected from MCTL region (Table 2).

Figure 5: Total Phenol content (mg/GAE dry wt.).

Catalase (CAT):

Catalase helps to rapidly neutralize hydrogen peroxide and prevent its harmful effects. The current study shows that the fruits of Artocarpus lakoocha collected from MCTL at its semi-mature stage recorded highest catalase enzymatic i.e., 1.216 U/ml. activity(Figure 6) followed by fruits of Mimusops elengi i.e., 1.1554 U/ml at its semi-mature stage collected from ESECP zone (Table 2).

Figure 6: Catalase enzymatic activity assay.

Peroxidase (POX):

Peroxiredoxins are a family of antioxidant enzymes that reduce hydrogen peroxide and organic peroxides, converting them into water and alcohols. They are involved in protecting cells from oxidative damage and also play a role in cell signaling.³ The study depicts that the fruit of Artocarpus lakoocha showed maximum Peroxidase enzymatic activity(Figure 7) i.e., 0.0246 ∆ OD /min/ g F.Wt. at its ripened stage collected from MCTL region followed by fruits of Mimusops elengi i.e., 0.0186 ∆ OD /min/ g F.Wt. at its matured stage collected from ESECP region of Odisha (Table 2).

Figure 7: Peroxidase Enzymatic Activity.

Super Oxide Dismutase (SOD)

Superoxide radicals are highly reactive and can cause cellular damage, by converting them into less harmful species like hydrogen peroxide, SOD helps protect cells from oxidative stress.³It is evident from the study that the ripened stage of the fruits of Mimusops elengi recorded highest SOD Enzymatic Activity i.e., 0.057 ∆ OD /min/ g F.Wt. However, for the fruit of Artocarpus lakoocha, the highest SOD activity (Figure 8) was recorded in semi-mature stage collected from MCTL region (Table 2).

Figure 8: Super Oxide Dismutase Enzymatic Activity.

Table 2: Quantitative analysis of Ascorbic Acid, DPPH% of Inhibition, FRAP assay, Total Phenol, CAT, POX, SOD.

Species name

Stages

AA^v

(mg/100 g F.Wt.)

AA^s

(mg/

100 g F.Wt.)

DPPH % of Inhibition

FRAP

(mM AEAC/

g dry wt.)

Total Phenol

(mg/GAE dry Wt.)

CAT

(U/ml)

POX

(∆ OD/ min/g F.Wt.)

SOD

(∆OD/Min/gm F.Wt)

AL^a

198.75±

4.595

132.16±

0.7

58.86

47.89±

1.09

66.58±

0.0901

1.046±

0.981

0.009±

1.023

0.031±

0.026

173.6±

4.31

120.09±

0.09

78.75

58.88±

1.87

97.86±

0.081

0.1554±

0.90

0.004±

1.009

0.076±

0.29

160.7±

3.979

99.35±

0.058

79.85

52.48±

2.02

215.34±

0.91

0.085±

0.671

0.0105±

1.302

0.057±

0.042

AL^b

183.5±

5.239

125.57±

0.5798

59.8

51.42±

0.98

40.13±

1.209

1.216±

0.872

0.001±

0.945

0.061±

0.23

158.72±

2.058

107.43±

0.081

73.559

60.95±

0.902

49.98±

1.18

1.024±

0.911

0.0052±

0.894

0.0108±

0.65

142.84±

3.66

81.90±

0.998

87.89

44.02±

1.023

289.44±

1.04

0.1054±

1.19

0.0246±

0.45

0.062±

0.45

ME^a

153.76±

4.895

111.16±

0.579

80.79

49.41±

2.201

47.31±

0.908

1.1554±

1.763

0.0027±

0.34

0.048±

0.22

138.88±

3.394

90.46±

0.102

70.48

63.17±

2.21

54.84±

0.879

0.878±

1.988

0.0186±

0.05

0.0784±

0.32

99.37±

3.457

60.34±

0.175

61.22

107.5±

1.089

115.74±

0.09

0.284±

2.01

0.005±

0.09

0.134±

0.7

ME^b

124±

2.3852

93.04±

0.74

68.12

41.49±

1.001

45.32±

0.12

0.9447±

1.233

0.0033±

0.149

0.0356±

1.002

84.91±

3.4587

46.2±

0.59

49.167

74.85±

1.08

399.14±

0.23

0.1748±

1.52

0.0012±

0.324

0.0267±

1.08

112.08±

3.241

56.86±

0.87

45.12

91.95±

1.022

147.27±

0.459

0.297±

1.98

0.0091±

0.12

0.0223±

1.97

N.B: “Data are expressed as Mean ± SD, (where n=5)”

Statistical significance was at p < 0.001 with compared to different stages of maturation of fruit along with Agroclimaticzones. “AL: Artocarpus lakoocha, ME: Mimusops elengi, S1: Semimature, S2: Mature, S3: Ripen, a: East and South Eastern Coastal Plain, b: Mid Central Table Land, v: Volumetric Method, s: Spectrophotometric Method, AA: Ascorbic acid”

DISCUSSION:

Reactive oxygen species (ROS) are molecules containing oxygen that have one or more unpaired electrons, or they may be oxygen-containing compounds lacking unpaired electrons, such as hydrogen peroxide (H₂O₂) and singlet oxygen (¹O₂). ROS, such as hydroxyl radicals or superoxide anions, can induce oxidative stress in cells, leading to damage to DNA, lipids, and proteins.²³ Apart from the culinary applications, the fruit of A. lakoocha has higher concentration of nutrients^36-38 and possesses a wide range of therapeutic qualities, including antibacterial, cytotoxic, anti-aging, anti-inflammatory, analgesic, anti-diarrheal, anti-glycation, and pancreatic lipase inhibitory activity.The fruits of the Mimusops elengi have cultural value and are employed in traditional medicine in the many regions where the species grows. The dried fruit powder is bisque in colour and coarse in texture. Earlier it was reported that Mimusops elengi has cytoprotective and antioxidant effects which could lead to use of herbal alternatives.³⁹ The study conducted by⁴⁰ from the methanolic extracts of the flower and leaves of Mimusops elengi has anti-inflammatory and antioxidant properties and those extracts showed a significant reduction in inflammation and demonstrated free radical scavenging abilities, suggesting potential therapeutic benefits for inflammatory conditions and oxidative stress-related disorders. However, the fruits of this plant have not been extensively studied so far. From this study, it was found that the variation in antioxidant capacities among both species could be attributed to the various agroclimatic conditions like temperature, humidity, soil condition, and rainfall.³⁴ It was evident from the analysis that the moisture content increases with the maturation of fruits.The moisture content for both the agroclimatic zones were found quite similar for M. elengi. Moisture content in fruits also affects its antioxidant potentiality of fruits which also influenced the phytochemical changes.³⁵ Ascorbic Acid is considered one of the most important immune enhancer compounds which can’t be synthesized by humans due to the lack of denovo synthesis of L- gulono- gamma- lactone oxidase in human body.⁴¹ Therefore, it is essential to intake a certain amount of daily dosage of Ascorbic Acid (Vitamin C) to maintain a healthy life. In this study for both species, the volumetric method was used; however to minimize the error spectrophotometric method was also followed. Ascorbic acid gets oxidized easily with a slight increase in temperature and the interference of light thus, becomes highly unstable and; was handled carefully. 10% Acetic Acid was used initially followed by the methods of ²⁹to get the extracted sample but it was not stable for more than 1 hour. Instead of that 3% Metaphosphoric acid was used to make it stabilized for at least 5-6 hours.³⁴ Notably there was variation in the Ascorbic acid content among both the species along with its stages of maturation and it was marked that the Ascorbic acid was found higher in the mature stages of both the species which may indicate the degradation of Ascorbic acid with the growth that may be affected by moisture and temperature.⁴² In the DPPH assay, jackfruit seeds showed moderate antioxidant potential with an IC50 value of 116.04μg/ml.⁴³ Phenolic compounds exhibit diverse antioxidant responses, which are influenced by their chemical structure.^44,45 However; there might be interference from other chemical components, like sugars or ascorbic acid.⁴⁵ These variations could stem from differences in genetic backgrounds, environmental conditions, or agricultural methods.^45,46An important oxidoreductase enzyme called “catalase (E.C. 1.11.1.6)” converts hydrogen peroxide into water and oxygen within the cells exposed to physiological stress, thereby reducing the harmful effects caused by free radicals. In both cancer and diabetic retinopathy, it is frequently employed.¹¹ Catalase is an enzyme that breaks down hydrogen peroxide (H₂O₂) into water (H₂O) and oxygen (O₂). Hydrogen peroxide is a byproduct of various metabolic processes and can be toxic in high concentrations. Peroxidase, the primary impact of the enzymes is on the flavor and colour retention of both fresh and processed fruits and vegetables.^47-49 Superoxide dismutase enzyme plays a vital role in defending cells from an assortment of illnesses, including cancer and heart attacks brought on by superoxide radicals. The enzyme superoxide dismutase (SOD) facilitates the conversion of superoxide radicals O^2- into O₂ and H₂O₂.⁵⁰ Furthermore, the enzymatic activities (SOD, CAT, POX) were notably higher in Artocarpus lakoocha fruits, suggesting its potential for enhancing cellular antioxidant defenses. The total phenolic content was most pronounced in Mimusops elengi at the mature stage collected from the MCTL zone, indicating its contribution to antioxidant capacity.Overall, the assessment of immune-enhancing properties in Artocarpus lakoocha and Mimusops elengi reveals valuable insights into their potential health benefits and antioxidant activities. Artocarpus lakoocha demonstrated significant levels of Ascorbic Acid, particularly at the semi-mature stage from the ESECP zone. On the other hand, Mimusops elengi exhibited strong antioxidant capabilities, with high DPPH scavenging and FRAP activity, especially at the semi-mature and ripen stages from the same ESECP zone. These findings underscore the importance of indigenous knowledge and traditional practices in identifying and utilizing wild edible fruits for their health-promoting properties. Over the long term, the insights gleaned from such research endeavors hold the potential to validate the therapeutic use of medicinal plants, thereby augmenting the intrinsic value of local botanical resources. This, in turn, could yield significant economic dividends, particularly for communities reliant on these natural assets. Future research should delve deeper into specific bioactive molecules credit worthy for these immune-enhancing attributes, facilitating their integration into functional foods or therapeutic formulations. Overall, Artocarpus lakoocha and Mimusops elengi hold promise as natural sources of antioxidants with potential benefits for human health and wellness.

CONCLUSION:

Artocarpus lakoocha showed high levels of Ascorbic Acid at the semi-mature stage belonging to the ESECP zone, while Mimusops elengi exhibited strong antioxidant activity, particularly at the semi-mature and ripen stages from the same zone. Artocarpus lakoocha also demonstrated elevated enzymatic activities (SOD, CAT, POX), while Mimusops elengi displayed the highest total phenolic content at the mature stage from the MCTL zone. In addition to helping with the food scarcity, increased utilization of these resources as these are high in antioxidant will benefit the local economy. Promoting research on indigenous plants is thus essential to preserve traditional knowledge of wild edible fruit species for future conservation and sustainable use, as it is being lost due to acculturation and the decline in plant biodiversity. To confirm their potential uses as natural antioxidant sources and to support their traditional uses in various medical practices, more research should focus on the fruit species broad in vivo antioxidant activities, link between individual compounds, antioxidants with various mechanisms, isolation and characterization of each compounds for antioxidant potencies.

ACKNOWLEDGEMENT:

The authors express their gratitude to the “Regional Plant Resource Centre, Bhubaneswar”, for providing funding for this research through state plan grant from the “Forest, Environment, and Climate Change Department, Odisha.” The assistance received from Tikarpada, Dhenkanal, and Nayagarh forest divisions during the sample collection, is highly acknowledged.

FUNDING:

State Plan Grant FY 2022-2023 (Forest, Environment and Climate Change Dept., Govt. of Odisha)

DECLARATION OF CONFLICTING INTERESTS:

No Conflict of interest.

ABBREVIATIONS:

AAE: Ascorbic Acid Equivalent; AEAC: Ascorbic Acid Equivalent Antioxidant Capacity; CAT: Catalase; DCPIP: 2,6- Di-chloro-phenol-indophenol; DNPH : 2,4- Dinitrophenylhydrazine; DPPH: 2,2- Diphenyl -1- picrylhydrazyl; EDTA: Ethylene diamine tetra-acetic acid; ESECP: East and South Eastern Coastal Plains; F.Wt. : Fresh Weight; FCR: Folin Ciocalteu Reagent; FRAP: Ferric Reducing Antioxidant Potential ; GAE: Gallic Acid Equivalent; MCTL: Mid Central Table Land; NBT: Nitro blue Tetrazolium Chloride; POX: Peroxidase; ROS: Reactive Oxygen Species; SOD: Super oxide dismutase; TPTZ: 2, 4,6-Tris(2-pyridyl)-s-triazine

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Received on 01.08.2024 Revised on 29.11.2024

Accepted on 28.02.2025 Published on 01.10.2025

Available online from October 04, 2025

Research J. Pharmacy and Technology. 2025;18(10):4679-4688.

DOI: 10.52711/0974-360X.2025.00673

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Fruit Species	ESECP			MCTL
Fruit Species	Semi-mature	Mature	Ripe	Semi-mature	Mature	Ripe
Artocarpus lakoocha	78.9	87.6	89.25	77.26	84.22	86.54
Mimusops elengi	72.5	79.12	76.24	72.44	79.42	78.51