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 Table of Contents  
Year : 2021  |  Volume : 6  |  Issue : 1  |  Page : 40-49

Diospyros exsculpta Buch.-Ham. an underutilized plant: Comparative determination of mineral, antimicrobial, antioxidant, and protein-binding qualities of different parts of plant

1 Central Ayurveda Research Institute, Central Council for Research in Ayurvedic Sciences (CCRAS), Kolkata, West Bengal, India
2 Central Ayurveda Research Institute, Central Council for Research in Ayurvedic Sciences (CCRAS), Jhansi, Uttar Pradesh, India
3 Regional Ayurveda Research Institute, Central Council for Research in Ayurvedic Sciences (CCRAS), Gwalior, Madhya Pradesh, India
4 National Ayurveda Research Institute for Panchakarma, Cherethurthy, Kerala, India
5 Regional Ayurveda Research Institute, Central Council for Research in Ayurvedic Sciences (CCRAS), Patna, Bihar, India
6 Central Council for Research in Ayurvedic Sciences (CCRAS), Jankapuri, New Delhi, India

Date of Submission21-Jul-2021
Date of Acceptance02-Aug-2021
Date of Web Publication14-Dec-2021

Correspondence Address:
Dr. Amit Kumar Dixit
Central Ayurveda Research Institute, CCRAS, 4 CN Block, Sec-5, Kolkata, West Bengal.
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jdras.jdras_9_21

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BACKGROUND: Diospyros exsculpta Buch.-Ham. (DE) belongs to the genus Diospyros, is an underutilized plant and needs to explore its phytochemical and biological activities. OBJECTIVES: The Present study aimed to compare phytochemical and biological activities such as antioxidant, antimicrobial, and protein-binding assay of twig and stem bark of DE. MATERIALS AND METHODS: Biological and phytochemical properties of three solvents extracts of twig or branches and stem bark were analyzed. Quantitatively, minerals, antioxidant qualities, total phenolic and flavonoid content, and protein binding were analyzed. Antimicrobial activities were assessed against four bacterial and three fungal strains. RESULTS: It was observed that stem bark and twig of DE showed a broad-spectrum antimicrobial activity. Good concentration of Cu, Fe, Mn, Na, P, and Zn has been observed. Comparatively, stem bark and twig have almost the same amount of nutrients (P ≤ 0.05). Value of total phenolic content (TPC) samples varied from 6.27 to 8.74 mg g-1 dry weight in methanol extracts, 18.48–24.09 mg g-1 DW in acetone extracts and 1.41-2.31 mg/g DW in petroleum ether extracts, which is called as gallic acid equivalents (GAEs). Total flavonoid content varied from 87.52 to 197.2 mg quercetin equivalent (QE)/g DW of sample for methanol extracts, 70.12–86.25 mg QE/g DW for acetone extracts, and 2.55–3.75 mg QE/g DW for petroleum ether extracts. CONCLUSION: It was observed that all three extracts of twig and stem bark of DE have significant variation for antioxidant activities. The antioxidant activities were variable from assay to assay as well as extract to extract. Overall, in antioxidant activities, polar extracts have shown the most effective results. All three extracts have shown a significant protein-binding assay, which was almost close to aspirin. Anti-inflammatory assay of various extracts has revealed potent anti-inflammatory activity. Significant linear correlation (P ≤ 0.05) of TPC with antioxidant activities favored its role in antioxidant activity. These comparative findings on phytochemical, biological activities of stem bark, and twig of DE provide compelling scientific evidence of its therapeutic usage.

Keywords: Anti-microbial, Diospyros exsculpta, phytochemicals, underutilized plants

How to cite this article:
Dixit AK, Kumar V, Meena AK, Srivastava B, Nair PG, Sahu D, Rotwar DS, Das M, Srikanth N. Diospyros exsculpta Buch.-Ham. an underutilized plant: Comparative determination of mineral, antimicrobial, antioxidant, and protein-binding qualities of different parts of plant. J Drug Res Ayurvedic Sci 2021;6:40-9

How to cite this URL:
Dixit AK, Kumar V, Meena AK, Srivastava B, Nair PG, Sahu D, Rotwar DS, Das M, Srikanth N. Diospyros exsculpta Buch.-Ham. an underutilized plant: Comparative determination of mineral, antimicrobial, antioxidant, and protein-binding qualities of different parts of plant. J Drug Res Ayurvedic Sci [serial online] 2021 [cited 2023 Feb 2];6:40-9. Available from: http://www.jdrasccras.com/text.asp?2021/6/1/40/332503

  Introduction Top

Natural product-based medicines are the basis of Western medicine and synthetic chemistry.Various reports mentioned that health-care systems in developing countries are based on these herbal drugs where 70%–80% population rely on natural products.[1],[2],[3] Approximately 70,000 species of plants are used in modern and traditional medicinal methods.[2],[4],[5] Still various plants in nature are underutilized and limited to tribes and need to be explored.[4],[5],[6],[7]

Diospyros exsculpta Buch.-Ham. (DE) belongs to the genus Diospyros and has limited international and national literature.[3],[7],[8] DE is limited to folklore drugs and needs to explore its phytochemical and biological activities. In the literature, two review articles have been noticed.[3],[7],[8] As per literature and folklore data, the whole plant of DE is used for health benefits w.r.t. food and medicinal benefits. The stem bark (StB) of DE is used in the treatment of wound’s healing by Indian tribals. Fruits of DE are used to cure dysentery and diarrhea, whereas roots of DE are active against scorpion venom.[3],[8] Hydroalcoholic (50%) extract of whole plant have shown cardiovascular activities. Hydroalcoholic (50%) extract of dry seeds has shown good antibacterial activities, respiration improvement quality, and blood control properties.[3],[7],[8] DE is part of Ebenaceae family having 500 species. The major chemical constituents of genus Diospyros are β-sitosterol, betulinic acid, betulin, lupeol, oleanolic acid, and ursolic acid.[3],[8]

Due to high nutritional value, the consumption of natural products or medicinal plants is increasing rapidly. These natural products or medicinal plants are used because of their medicinal and antioxidant importance.[3],[8],[9],[10],[11] These natural products are rich of phytochemicals such as polyphenols and flavonoids, so they are considered as alternate of synthetic products having various health hazards.[2],[3],[8],[12] Moreover, metal elements are very important for whole organisms w.r.t. cellular activities. Na and K balance the osmotic pressure regulations.[3],[12] Fe, Co, and Cu are very important to maintain hemoglobin, blood transport, functioning of proteins, and various metalloenzymes of body.[2],[3],[8],[12] Nature and natural sources are considered to be the best option for essential metal ions. More significantly, plants are the integral source of civilization to provide the minerals and antioxidants.

In this study, we have explored the underutilized plant DE. To the best of our knowledge, this is the first detailed study on Diospyros exsculpta and its phytochemical and biological activities such as antimicrobial, antioxidant, and protein-binding assay of the StB and Tw of DE.

  Materials and Methods Top

Plant material collection

The plant materials (Tw and StB) of DE were collected from the local area, District Gwalior of Madhya Pradesh, India. The plant materials were identified and authenticated through standard taxonomical procedure. Tw and StB were dried under dark conditions. The shaded dried samples of Tw and StB were powdered, dried, and stored in an air-tight zip-lock polyethylene for extraction of solvent. The drugs were dried and kept at 25°C at room temperature.

Determination of physicochemical parameters

The Ayurvedic Pharmacopoeia of India (API) protocols were followed to determine the physicochemical parameters (foreign matter, total ash, pH, extractive values [alcohol/water], acid insoluble ash, and its loss on drying).[13] Phytochemical analysis was done as described in the literature.[4],[9],[12]

Preparation of plant extracts

Three different solvents viz. acetone, methanol, and petroleum ether were selected and used to prepare the extract. To extract the phytoconstituents, the extracts were prepared as per API procedure. Briefly, 4 g of the dried powdered samples of Tw and StB of DE were dipped in 250 mL of conical flask having 100 mL of solvent. Flasks were shaken for 6 h. After 24 h, samples of plant material were filtered via Whatman No. 1 filter paper and filtrate was used for further analysis.

Sample preparation for metal ion analysis

The dried and well-powdered samples were microwave digested (ETHOS 900 Microwave System, Milestone, Germany).[14],[15],[16] In triplicates, 25 mg of the samples were weighed in microwave digester vessels. After that 1:2 mixture of HNO3 and H2O2 were added and the samples were kept in digestion unit. After digestion, each sample was diluted with distilled water to make up the solution 15 mL and analyzed by using Inductive Coupled Plasma Optical Emission Spectroscopy (ICP-OES) (Optima 8000, Perkin Elmer, Inc. Shelton, CT, USA).

Metal analysis using Inductive Coupled Plasma Optical Emission Spectroscopy

The ICP-OES instrument is known for multielemental analysis with high accuracy and high range of linearity.[17],[18],[19] The details of instrumentation were as follows: operating conditions: viewing: geometry axial; RF power 1.0 kW; replicate reading time: 9.0 s Argon gas flow rate: 15.0 L min−1; nebulizer gas flow rate: 0.75 L min−1; and auxiliary gas flow rate: 1.50 L min−1. The operating wavelength of selected element was as follows: Mg (285.213 nm), Al (396.153 nm), Na (589.592 nm), P (213.617 nm), K (766.490 nm), Cr (334.940 nm), Ca (317.933 nm), Ti (267.716 nm), Fe (238.204 nm), Mn (257.610 nm), Cu (327.393 nm), Ni (231.604 nm), Co (228.616 nm), Zn (206.200 nm), Cd (228.802 nm), As (193.696 nm), and Ag (328.068 nm). All samples were analyzed in triplicates.

Antioxidant, anti-inflammatory total flavonoids content, total phenolic content, and protein-binding assays using ultraviolet–visible spectrophotometric methods

In this study, total flavonoids content (TFC) and total phenol content (TPC) were analyzed by using a ultraviolet–visible spectrophotometer (Shimadzu-1800).[4],[5],[6] The antioxidant capacity was analyzed using various assays viz. metal chelating (MC), ferric reducing (FR), reducing power (RP), superoxide anion (SA), and nitric oxide (NO) scavenging activity.[1],[4],[5],[6],[9],[12] To analyze the protein-binding and anti-inflammatory activities, literature methods were used.[1],[20]

Antimicrobial assays

To analyze the antimicrobial activities, four bacterial and three fungal strains were used and purchased from the NCIM Pune and MTCC Chandigarh. The fungal strains were as follows: Aspergillus parasiticus NCIM 696, A. niger NCIM 501 and Candida albicans MTCC 183. The bacterial strains were as follows: Pseudomonas aeruginosa MTCC 4673 (Gram –ve),  Salmonella More Details enteric MTCC 164 (Gram –ve), Staphylococcus aureus MTCC 7443 (Gram +ve), and Streptococcus mutans MTCC 497 (Gram +ve). To analyze the antimicrobial activities, previously described and standardized protocols were followed.[1],[21],[22]

Statistical analysis

All the quantitative tests of phytochemical extraction were carried out in triplicates. Analysis of variance (ANOVA) at P ≤0.05 and other statistical analysis were performed using SPSS version 16.

  Results Top

Physicochemical parameters

In this study, the Tw and StB of DE have shown pH values 6.10 ± 0.12 and 6.20 ± 0.16, respectively. The experimental values of total ash of Tw and StB were 10.17 ± 0.74% and 9.91 ± 0.56%, respectively. The observed values of acid-insoluble ash for the Tw and StB of DE were 0.81 ± 0.03% and 0.79 ± 0.03%, respectively. The observed values for water-soluble ash for the Tw and StB of DE were 2.13 ± 0.21% and 2.43 ± 0.24%, respectively. The experimental loss on drying for the Tw and StB of DE were 0.214 ± 0.05% and 0.198 ± 0.07%, respectively. The experimental extractive values for the Tw and StB of DE in petroleum ether were 4.81% and 5.01%, respectively. The observed extractive values for the Tw and StB of DE in acetone were 12.40% and 12.43%, respectively. The observed extractive values for the Tw and StB of DE in methanol were 32.18% and 31.65%, respectively. Of three extraction solvent used under this study, a significant yield was found in the methan and acetone as compared to that of petroleum ether extract.

Qualitative analysis of phytochemicals

Qualitative results of phytochemicals of Tw and StB of DE are tabulated in [Table 1]. Good variety of phytochemicals has been found in the acetone extract as compared to that of other two extract viz. methanol and petroleum ether [Table 1]. [Table 1] shows the presence of terpenoids in all three extracts, of Tw and StB of DE.
Table 1: Qualitative analysis of phytochemicals present in the various extracts of stem bark and twigs/branches of Diospyros exsculpta

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Essential metal ion content

An effort was made in this study to find out content of metal ions such as Ca, Mg, Na, K, Cr, Mn, Fe, Cu, and Zn in Tw and StB of DE by using ICP-OES technique. In ICP-OES results, metal ions, such as Fe, Ca, Zn, Cu, Na, and Mg, have been found in Tw and StB of DE at good concentration (P ≤ 0.05) [Table 2].
Table 2: Minerals and heavy-metal ions of small branches and stem bark of Diospyros exsculpta

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Heavy metal content

[Table 2] shows the observed value of Hg, Pb, Cd, and As within API and WHO prescribed limits. Although the quantification of heavy-metal ions of medicinal plant is important, maximum time it is avoided for single study.

In vitro antioxidant assays

In this study, a group of the antioxidant assay was performed where six assays were used [Table 3] and [Figure 1]. In DPPH assay, the observed values of antioxidant radical scavenging (RS) activity (%) of petroleum ether, acetone, methanol extracts of Tw and StB of DE were 8.46 ± 0.95%, 5.88 ± 0.78%, 88.71 ± 2.67%, 90.12 ± 3.76%, 86.96 ± 2.73%, and 90.36 ± 2.77%, respectively. Results indicate that acetone, methanol extracts of Tw, and StB of DE have close value of antioxidant property as that of ascorbic acid (91.30% ± 2.78%) [Figure 1A].
Table 3: In vitro antioxidant assays of the various extracts of Diospyros exsculpta Buch.-Ham.

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Figure 1: In vitro antioxidant assays of the various extracts of Diospyros exsculpta. (A) DPPH radical scavenging activity. (B) Metal chelating scavenging activity. (C) FRAP scavenging activity. (D) Reducing power. (E) Superoxide radical scavenging activity. (F) Nitric oxide radical scavenging activity. Code: A = small branches, B = stem bark. Solvents: petroleum ether (1), acetone (2), and methanol (3). Values are expressed as mean ± standard deviation (SD) (n = 3).

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In the second assay (RP), antioxidant values (%) for three extracts viz. petroleum ether, acetone, and methanol extracts of Tw and StB of DE were 81.70 ± 0.74%, 55.67 ± 0.65%, 85.52 ± 0.83%, 93.84 ± 0.94%, 83.83 ± 0.74%, and 63.54 ± 0.53%, respectively. Here all extracts of Tw and StB of DE have shown a significant chelating or antioxidant property as compared with that of ethylenediaminetetraacetic acid (EDTA) or control (89% ± 0.84%) [Figure 1B]. [Figure 1B] shows that acetone extract of StB of DE has more activity than that of control (89 ± 0.84%), which was 93.84 ± 0.94% [Figure 1B].

In the third assay of antioxidant, that is in FRAP antioxidant activity, the observed antioxidant value was 82.73 ± 0.42% with ferrous sulfate as control. For three extracts viz. methanol, acetone, extracts of Tw and StB of DE the FRAP antioxidant values were 57.14 ± 0.23%, 46.02 ± 0.19%, 35.88 ± 0.24%, and 20.16 ± 0.17%, respectively. [Figure 1C] shows that methanol and acetone extracts of Tw and StB of DE have mild antioxidant property, whereas petroleum ether extracts have no FRAP antioxidant activities as compared to control [Figure 1C].

In the fourth assay (RP), antioxidant activity ascorbic acid was selected as control and the observed experimental antioxidant capacity was 33.23 ± 0.44%. [Figure 1D] shows that the petroleum ether, acetone, and methanol extracts of Tw and StB of DE have 2.40 ± 0.07%, 2.0 ± 0.05%, 26.71 ± 0.32%, 20.82 ± 0.22%, 21.79 ± 0.24%, and 14.11 ± 0.13% RP antioxidant activity. It is found that under RP antioxidant activity methanol and acetone extracts of Tw and StB of DE have almost the same antioxidant property as that of control one, that is, ascorbic acid [Figure 1D].

The fifth antioxidant assay was the superoxide radical scavenging (SRC) activity assay. The observed SRC antioxidant capacity for ascorbic acid (control) was 64.98 ± 0.38%. Under same conditions, values for petroleum ether, acetone, and methanol extracts of Tw and StB were 18.38 ± 0.17%, 27.91 ± 0.19%, 47.81 ± 0.56%, 47.41 ± 0.53%, 43.71 ± 0.34%, and 38.98 ± 0.29%, respectively. It has been found that polar solvents such as acetone and methanol have SRC antioxidant capacity [Figure 1E].

In the sixth antioxidant assay, RS assay was used and the observed value for ascorbic acid (control) was 29.21 ± 0.58%. Under same conditions, values for petroleum ether, acetone, and methanol extracts of Tw and StB of DE were 23.40 ± 0.17%, 22.88 ± 0.11%, 17.84 ± 0.61%, 17.49 ± 0.32%, 21.48 ± 0.23%, and 16.48 ± 0.22%, respectively. Clearly, nonpolar extracts have very good antioxidant property polar extracts of Tw and StB of DE as compared with control [Figure 1F].

Total flavonoids and phenol contents

The results of TPC and TFC of herbal drugs under study are shown in [Table 3]. It has been noticed that acetone extracts have good values of TFC and TPC. The exact order of TFC and TPC (for both Tw and StB of DE) was as follows: acetone > methanol >> petroleum ether. Except for methanol extract, no significant difference was observed for TPC of Tw and StB of DE. A significant difference was observed for TFC of Tw and StB of DE except for petroleum ether extracts. The TPC was calculated as equivalents of gallic acid (GAE) (Y = 0.303X + 0.007), R2 = 0.999, as shown in [Table 3]. The results for the TPC in extracts, expressed as equivalents of gallic acid (GAE)/g DW of plant, varied to a great extent and ranged from 0.011 to 2.25 mg GAE/g crude extract in petroleum ether extracts, 18.48–24.09 mg GAE/g DW in acetone extracts and 6.27–8.74 mg GAE/g DW extract in methanol extracts [Table 3]. Similarly, TFC expressed as quercetin equivalents (QE) (Y = 0.040X + 0.012), R2 = 0.999 is depicted in [Table 3]. The results for the TFC in extracts, expressed as QE/g DW of plant, varied to a great extent and ranged from 2.55 to 3.75 mg QE/g DW in petroleum ether extracts, 70.12–86.25 mg QE/g DW in acetone extracts and 87.52–197.2 mg QE/g DW in methanol extracts [Table 3].

Anti-inflammatory and protein-binding assays

The results of anti-inflammation activity were checked by using the denaturation of proteins assay. The acetylsalicylic acid was used as control and the results of three extracts under study were compared with that of control. Overall significant inhibitory results ranging from 95% to 98% were found in the heat-induced recovery of BSA protein. Here, the methanolic extract has shown good results as compared to other two solvents viz. acetone and petroleum ether extracts. The BSA protein binding assay of Tw and StB of DE is shown in [Table 3], where acetylsalicylic acid was used as a control [Table 3].

Antimicrobial activities

[Table 4] shows that all three extracts have shown a diverse range of antibacterial capacity due to the presence of a wide class of phytochemicals as mentioned in [Table 2]. In this study, polar extracts viz. methanol and acetone were more effective as compared to nonpolar petroleum ether extracts. It has been found that Tw and StB of DE extracts showed a broad-spectrum of antibacterial activity in both polar. [Table 4] shows maximum inhibition of S. mutans caused by the polar solvents of Tw and StB of DE. Acetone extract has significantly inhibited the growth of all the strains under study. Methanolic extract has significantly inhibited the growth of P. aeruginosa and S. mutans.
Table 4: Antimicrobial activities of various extracts of Diospyros exsculpta of small twigs/branches and stem bark

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In the case of natural products, there is a direct correlation between pH and presence of phytochemical constituents. Phytochemicals such as phenolic and flavonoids can show pH between 4 and 7 and show the high antioxidant activity.[4],[5],[12] The experimental results of the physicochemical parameters of Tw and StB of DE were almost similar, which has shown equal utility of Tw and StB. These results may highlight the presence of same types of phytochemicals in the Tw and StB of DE. Consequently, Tw may be used as alternate of StB after biological tests and dose determination.[4],[5],[6],[7]

It is the well-known fact that variation among the phytochemicals among same plant depends upon the geographical conditions.[4],[5],[6],[7] Metal ions, especially essential metal ions, are an important part of human life.[23] Nature has given a beautiful gift to human races where each plant has a different kind of essential metal ions. Among the plants, medicinal plants are the special gift where metal ions are available with phytochemicals and play a vital role in the potency of any plant.[16],[24] If we consider the World Health Organization (WHO) reports, billions of population are suffering from deficiency of essential metal ions like Zn and Fe.[23],[25] The second most important aspects is the heavy-metal limit for herbal drug where each herbal drug has to comply as per international standards. In this study, heavy metals, such as Cu and Cr, are within permissible limits of WHO, that is, 2 mg/kg and 150 mg/kg, respectively.[6],[24],[26],[27] The metal ion Cr is beneficial to diabetes therapy, but higher concentration of Cr may cause diseases such as cancer and respiratory problems.[24],[28] Similarly, Cu metal ion is involved in maximum physiological process, but high diet or intake of Cu-rich diet harms kidney, liver, gastrointestinal system, and even may cause the death.[24],[29] It is well-known metal ions, such as Zn, Co, and Fe, are basis of maximum physiological process and are very vital for living beings.[23],[25]

In the world market, Indian Medicinal System is still not as popular as expected due to lack of proper quality control data in the shape of contemporary frame.Recently various authors have reported the presence of heavy-metal ions such as Hg, Pb, Cd, and As more than WHO prescribed limit.[24],[28],[30] The other conditions such as agriculture soil and water used for irrigation process are responsible for higher limits of heavy metals for which WHO has also issued the guidelines on Good Agricultural Practices for herbal drugs.[24],[26],[30]

Medicinal plants are good resources of antioxidants and medicinal properties. Both properties viz. antioxidants and medicinal potency are related to each other.[4],[5] A single antioxidant assay is never a good a practice to find out the exact antioxidant capacity of any natural compound.[6],[12],[22],[24],[31],[32],[33],[34],[35],[36],[37],[38],[39],[40] In the present results, all three extracts have shown antioxidant potential where polar solvents such as acetone and methanol have shown dominancy in all assay. These antioxidant-rich extract products/compounds are potent against various illnesses/conditions such as cancer, inflammation, fatigue, and weakness. The naturally occurring antioxidant-rich diet is a good replacement over hazardous synthetic antioxidants viz. butylated hydroxyl anisole, and propyl gallate.[4],[5]

Designing and formulation of herbal drug is based on high phenolic and flavonoids content of aromatic plant(s). The presence of a high percentage of flavonoids and phenolic content reveals the ability of the plant to treat multiple diseases.[12],[31] The data was analyzed using linear correlation to find the correlation between the antioxidant activities to TFC and TPC of twig and StB of DE plant. Relatively strong linear positive association of acetone and methanolic extracts was observed among the MC assay, DPPH assay, FRAP assay, RP assay, superoxide anion free radicals scavenging, and molybdenum assay percentage, which signifies the role of flavonoid and phenol compounds as reducing agents that contribute in antioxidant activity. However, a very weak linear correlation between NO radicals scavenging percentage of sample to phenolic compounds was observed, which could be due to variation in the action mechanism of the assays and nature of the plants under investigation.[31],[32],[33],[34],[35],[36],[37],[38],[39],[40] However, from [Table 3], it is apparent that the plant under investigation has significant phenol and high flavonoid content in acetone and methanol extracts showed notable antioxidant potential indicating the good potency of its various compounds.[12],[31] The anti-inflammatory activity were also assessed by using the denaturation of proteins which is an important aspect for drug delivery.[20]

Our results indicated that all the strains under study were resistant to most of the petroleum extracts, whereas S. enteric and Staphylococcus aureus were resistant to most of the methanolic extracts. This proves the role of extra outer membrane present in the cell selectively permeates the passage of samples. Same time plant under study showed the inhibition of above-mentioned bacteria as it was containing dynamic phytochemicals which can act as the inhibitor of bacterial development deprived of penetrating inside the cell itself. Results of the antifungal experimentation are depicted in [Table 4]. It was found that all phytochemicals extracts have significantly inhibited the growth of fungal strain A. parasiticus. However, petroleum extract was nonactive against the fungal strains C. albicans and A. niger. Methanolic and acetone extracts were active against the fungal strains A. niger and C. albicans. This confirms that alkaloids and flavonoids were present in all extracts, which showed the antimicrobial activity due to the presence of metabolites. Our results were in agreement with literature related to plant extracts showing antimicrobial activities, which is due to the presence of effective phytochemical compounds viz. flavonoids, tannins, and alkaloids.[31],[32],[33],[34],[35],[36],[37],[38],[39],[40]


The present results deliver the supporting evidence for the therapeutic use of phytochemicals extracted from selected plants. In this study, methanolic and acetone extracts showed far better activity in all the phytochemical assays apparently due to efficient extraction of phytochemicals. A significant relationship between the phenolic compounds and antioxidant capacities indicated that these are the main contributors to the antioxidant capacities of these selected plants. Present findings pave the way to isolate exact compounds having bioactive properties and commercially valuable using suitable plant parts on the basis of their good therapeutic values and suitable solvent for extraction. It is worthy to say that both parts of DE have shown very good minerals compositions, and antioxidant and antimicrobial properties. Further, there is a need to explore the studies viz. isolation and antioxidant biological properties of this underutilized plant.

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Conflicts of interest

There are no conflicts of interest.

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Kumar V, Singh S, Singh R, Upadhyay N, Singh J. Design, synthesis, and characterization of 2,2-bis(2,4-dinitrophenyl)-2-(phosphonatomethylamino)acetate as a herbicidal and biological active agent. J Chem Biol 2017;10:179-90.  Back to cited text no. 21
Kumar V, Kaur S, Singh S, Upadhyay N. Unexpected formation of N’-phenyl-thiophosphorohydrazidic acid O,S-dimethyl ester from acephate: Chemical, biotechnical and computational study. 3 Biotech 2016;6:1.  Back to cited text no. 22
Long JK, Banziger M, Smith ME, Diallel E. Analysis of grain iron and zinc density in southern African-adapted maize inbreds. Crop Sci 2004;44:2019-26.  Back to cited text no. 23
Mulaudzi RB, Tshikalange TE, Olowoyo JO, Amoo SO, Plooy CPD. Antimicrobial activity, cytotoxicity evaluation and heavy metal content of five commonly used South African herbal mixtures. S Afr J Bot 2017;112:314-8.  Back to cited text no. 24
Alloway BJ. Zinc in Soils and Crop Nutrition. 2nd ed. Brussels, Belgium and Paris, France: International Zinc Association (IZA), IFA; 2008.  Back to cited text no. 25
WHO Guidelines on Good Agricultural and Collection Practices (GACP) for Medicinal Plants World Health Organization. Geneva: WHO; 2003.  Back to cited text no. 26
WHO Guidelines for Assessing Quality of Herbal Medicines with Reference to Contaminants and Residues Switzerland. Geneva: WHO; 2007.  Back to cited text no. 27
Agency for Toxic Substances and Disease Registry, 2012 Toxicological Profile for Chromium Public Health Services. Atlanta, GA: US Department of Health and Human Services.  Back to cited text no. 28
Agency for Toxic Substances and Disease Registry (ATSDR). Toxicological Profile for Copper Public Health Services. Atlanta, GA: US Department of Health and Human Services; 2004.  Back to cited text no. 29
Stahl T, Taschan H, Brunn H. Aluminium content of selected foods and food products. Environ Sci Eur 2011;11:23-37.  Back to cited text no. 30
Dehghan H, Sarrafi Y, Salehi P. Antioxidant and antidiabetic activities of 11 herbal plants from Hyrcania region, Iran. J Food Drug Anal 2016;24:179-88.  Back to cited text no. 31
Kumar V, Singh SB, Singh S. COVID-19: Environment concern and impact of Indian medicinal system. J Environ Chem Eng 2020;8:104144.  Back to cited text no. 32
Kumar V, Singh S, Singh A, Subhose V, Prakash O . Assessment of heavy metal ions, essential metal ions, and antioxidant properties of the most common herbal drugs in Indian Ayurvedic hospital: For ensuring quality assurance of certain ayurvedic drugs, Biocatal. Agric Biotechnol 2019;18:101018.  Back to cited text no. 33
Kumar V, Singh S, Singh R. Phytochemical constituents of guggul and their biological qualities. Mini Rev Org Chem 2020;17: 277-88.  Back to cited text no. 34
Kumar V, Singh S, Bhadouria R, Singh R, Prakash O . Phytochemical, analytical and medicinal studies of Holoptelea integrifolia roxb planch: A review. Curr Tradit Med 2019;5:270-7.  Back to cited text no. 35
Kumar V, Singh S, Srivastava B, Patial PK, Kondalkar SA, Bharthi V. Volatile and semi-volatile compounds of Tephrosia purpurea and its medicinal activities: Experimental and computational studies. Biocatal Agric Biotechnol 2019;20:101222.  Back to cited text no. 36
Kumar V, Singh S, Kondalkar SA, Srivastava B, Sisodia V, Bharthi V . High resolution GC/MS analysis of the Holoptelea integrifoli’s leaves and their medicinal qualities. Biocatal Agric Biotechnol 2019;22:101405.  Back to cited text no. 37
Kumar V, Singh S, Srivastava B. Bhadouria R, Singh R. Green synthesis of silver nanoparticles using leaf extract of Holoptelea integrifolia and preliminary investigation of its antioxidant, anti-inflammatory, antidiabetic and antibacterial activities. J Environ Chem Eng 2019;7:103094.  Back to cited text no. 38
Kumar V, Singh S, Singh S, et al. Methods and techniques for the chemical profiling and quality control of natural products and natural product-derived drugs. In: Singh J, Meshram V, Gupta M, editors. Bioactive Natural Products in Drug Discovery. Singapore: Springer; 2020. p. 585-98.  Back to cited text no. 39
Kumar V. Phytochemical, pharmacological activities and ayurvedic significances of magical plant Mimosa pudica Linn. Mini Rev Org Chem 2012;18:296-312.  Back to cited text no. 40


  [Figure 1]

  [Table 1], [Table 2], [Table 3], [Table 4]


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