Antioxidant Activity of Flavonoids and Phenolic Acids from <i>Dodonaea angustifolia</i> Flower: HPLC Profile and PASS Prediction

Tessema, Fekade Beshah; Gonfa, Yilma Hunde; Asfaw, Tilahun Belayneh; Tadesse, Mesfin Getachew; Bachheti, Rakesh Kumar

doi:https://doi.org/10.1155/2023/8315711

Journal of Chemistry

On this page

Abstract Introduction Materials and Methods Results Discussion Conclusions Data Availability Conflicts of Interest Acknowledgments Supplementary Materials References Copyright Related Articles

Special Issue

Natural Bioactive Compounds: Chemistry, Extraction, and Applications

View this Special Issue

Research Article | Open Access

Volume 2023 | Article ID 8315711 | https://doi.org/10.1155/2023/8315711

Antioxidant Activity of Flavonoids and Phenolic Acids from Dodonaea angustifolia Flower: HPLC Profile and PASS Prediction

Fekade Beshah Tessema,^1,2Yilma Hunde Gonfa,^1,3Tilahun Belayneh Asfaw,^1,4Mesfin Getachew Tadesse,^1,5and Rakesh Kumar Bachheti^1,5

Academic Editor: Marwa Fayed

Received04 Nov 2022

Revised23 Nov 2022

Accepted19 Dec 2022

Published30 Mar 2023

Abstract

Background. Dodonaea angustifolia is a known medicinal plant across East Arica. The flower of D. angustifolia is not well investigated in terms of phytochemistry and biological activities. This study aims to investigate the presence of flavonoids and phenolic acids in the flower of D. angustifolia and its antioxidant activity. Methods. Preliminary phytochemical screening was carried out using the standard protocols. Antioxidant activity evaluation using DPPH assay and total phenol content (TPC) and total flavonoid content (TFC) determinations in the flower extract were compared with the values of the leaf extract. UHPLC-DAD analysis was managed to develop the profile of the flower extract. Prediction of biological activity spectra for substances (PASS) was done using an online server for antioxidant and related activities. Results. Preliminary phytochemical screening and TPC and TFC values confirmed the presence of flavonoids and phenolic acids. From the HPLC analysis of flavonoids, quercetin, myricetin, rutin, and phenolic acids such as chlorogenic acid, gallic acid, and syringic acid were detected and quantified. The biological activity spectrum was predicted for the detected and quantified polyphenols. Conclusions. D. angustifolia flower is a rich source of flavonoids and phenolic acids, which are extractable and can be checked for further biological activity. It was possible to identify and quantify phenolic compounds through HPLC analysis in the methanol extract of D. angustifolia flower. The PASS biological activity prediction results showed that there were stronger antioxidant activities for the identified flavonoids. Future work will emphasize the isolation and characterization of active principles responsible for bioactivity.

1. Introduction

Dodonaea angustifolia Lf (syn: Dodonaea viscosa) is a species belonging to the family Sapindaceae and known for many therapeutic purposes [1]. Locally, in Ethiopia, it is known by the names such as Karkare (Agew), Kitkitta (Amh, Gur), Termien (Geez), Ettecca (Oro), Intanca (Sid), Tahses (Tre, Tya), and Den (Som) [2, 3]. The plant is an erect bushy shrub 5–8 m high, with simple and alternate leaves, yellowish-green small flowers stacked without petals, yellowish-green capsule fruits, and flowering after the rainy season from August to September [3]. It occurs in most parts of Ethiopia and is pantropically known for fast-growing and used for soil stabilizing and reforestation activities [1].

Ethno-medicinal reports showed that parts of this plant are known to cure different human and cattle ailments. The roots are used for toothache and wound healing [4], parasitic worms [5], and tapeworms [6, 7]. Roots with leaves have been used for trachoma [8]. Leaf decoctions, juice, and extracts of D. angustifolia are used for the treatment of taeniasis [9], liver ailment [10], wound healing [8, 11, 12], eye infection, herps and fire burn [13], malaria [12, 14], cancer [15], and skin infection and wound [16–18]. The leaves are also used externally for itchy skin and as a remedy for skin rashes [19]. It is also reported that the leaf extract is known for mild purgative and soared throat [17] and hemorrhoid [20]. As an ointment for head swelling, bursting, and hair fungus, dried, powdered leaves and a paste made of oil have been employed [12].

A large group of phytochemicals have been reported from Dodonaea species. Melaku et al. isolated pinocembrin (flavanone), santin (flavanol), and clerodane diterpenes using bioassay-guided extraction and chromatographic separation [21] from D. angustifolia. Similarly, Omosa et al. reported flavonoids (3,4′,5,7-tetrahydroxy-6-ethoxyflavone, 5-hydroxy-3,4′,7-trimethoxyflavone, isokaempferide, kumatakenin, rhamnocitrin, and diterpenoids ((ent-3β,8α)-15,16-epoxy-13(16),14-labdadiene-3,8-diol, 2β-hydroxyhardwickiic acid, and dodonic acid) from the leaf extract of same species using chromatographic separation [22]. Methoxymkapwanin and Mkapwanin were also isolated from the leaf surface exudate using serial extraction and column chromatography fractionation [23].

From earlier investigations, both crude methanol extract [24] and quercetin derivatives [22, 25, 26] isolated from D. angustifolia showed antibactericidal efficacy against gram-positive and gram-negative bacteria. The aqueous extract demonstrated analgesic and antipyretic potential in mice and rats [27]. Ethanol extract displayed a combination of antioxidant and antiproliferative properties with little to no damage to normal cells [28, 29]. Nonpolar extracts of the leaves of D. angustifolia inhibited the viral growth at subtoxic concentrations [30]. Crude extract of both the leave [31, 32] and root [33] showed the strong antiplasmodial activity against P. berghei infected mice. This is in agreement with its medicinal use traditionally to treat malaria [34]. The antihelminthic property of D. angustifolia was reasoned out for the polyphenols present, including kaempferol, quercetin, and myricetin-based flavanol [35].

D. angustifolia is a medicinal plant frequently used to treat toothache, microbial infections, and fever [36]. It showed antifungal activities and was found to be nontoxic. It is also used with other medicinal plants for musculoskeletal ailments [37] and bone fracture [6] with a higher informant agreement ratio.

The biological activity spectrum predicts many different compounds’ biological activity types. Since it is solely dependent on the compound’s structure, it is regarded as an intrinsic property of the substance [38]. The multilevel neighborhoods of atoms (MNA), which are original descriptors, are used in PASS (prediction of activity spectra for substances) to characterize chemical structure [39]. A MOL or SDF (structure data file formats) with the structural details of the molecules being studied serves as the input data for PASS. MNA descriptors are generated automatically using the data from the input files. Using data from MNA descriptors for both active and inactive compounds, for each activity, two probabilities are computed: Pa is the probability that a compound is active, while Pi is the probability that a compound is inactive. Pa and Pi have values that range from 0.000 to 1.000 since they represent probabilities (with 3 appropriate decimals determined) and Pa + Pi < 1 since these probabilities are computed independently. Pa and Pi can be thought of as measurements of the substance being studied that fall within the categories of active and inactive substances, respectively [38].

The antioxidant action of flavonoids and phenolic acids includes suppressing the formation of reactive oxygen species by inhibiting the respective enzymes, scavenging free radicals, and triggering antioxidant defence [40]. These phytochemicals also protect the disintegration of the lipid of biomembrane by preventing or hindering lipid peroxidation [41]. Flavonoids are synthesized by plants following a microbial infection and are known to possess antioxidant and antibacterial activities. The mechanism and activity level depends on the polyphenols’ specific structural variety [41]. Flavonoids have a more significant number of physiological activities promoting human health and minimizing the risk of being infected by a broad spectrum of pathogens.

Flowers of D. angustifolia are ideal for bee forage and are considered a major agricultural value of the plant [3]. The seasonal flower of this plant is not well investigated in terms of phytochemistry and biological activities. Other parts including leaves, seeds, stem bark, and roots were investigated for their phytochemical constituents [22, 23, 26] and biological activities [21, 22, 25, 27, 31, 33, 34].

This study aimed to investigate the profile and antioxidant activity of the flower of D. angustifolia and compare it to the plant’s leave. For this purpose, HPLC analysis and PASS online prediction of biological activities were used.

2. Materials and Methods

2.1. Chemicals and Reagents

All the extraction chemicals and reagents used for the total content of phenol and flavonoid determination were of AR grade. While for HPLC analysis and the antioxidant activity evaluation, HPLC grade solvents and reagents were used. Water was distilled and purified by MQ (18.2) at 21°C in a water purification system (Purelab flex 4 Elga). Phenolic acid standards: syringic acid, chlorogenic acid, and gallic acid; flavonoid standards: myricetin, quercetin, rutin, and kaempferol references purchased from Sigma (>99.9%, Sigma, China). 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) for antioxidant test and ascorbic acid are obtained from Sigma (>99.9%, Sigma, China).

2.2. Plant Material Collection and Pretreatment

The flower part of D. angustifolia was collected from Addis Ababa Science and Technology University campus. Ato Melaku Wondafrash identified the plant and a herbarium sample was deposited at the national herbarium (voucher number: FB-001/11) in the College of Science, Addis Ababa University, Ethiopia. Following collection, the samples were cleaned with tap water and then distilled water to get rid of dirt and other debris. Then, the samples were chopped into smaller sizes and spread onto clean polyethylene plastic sheets at room temperature (23 ± 3°C). The air-dried samples were ground using a sample grinder (stainless steel 700 g electric grains, spices, herbs, cereals, and dry food grinding mill, China).

2.3. Ultrasonic Assisted Extraction (UAE)

The extracts of air-dried and blended leaves and flower samples of D. angustifolia (5 g of each) were obtained from an intelligent ultrasonic processor (SJIA-950W, probe 6) sonicator in 25 mL methanol. The method optimization of UAE followed a method reported by Zakaria et al. [42], with minor modifications considering the bioactive components we are dealing with. Briefly, the settings for sonication were temperature 35°C, time 15 min, and power rate 50%. Up on the optimized method suggested by Zakaria et al. (2021), the nature of the solvent (methanol) was expected to compromise the polarity difference between the phenolic acid and flavonoid with varied polarity. The temperature is also reasonable for the extraction of bioactive components. After 2x successive sonication for each aliquot extract, the extracts were centrifuged using proanalytical @ 600 (10x) for 20 min. Whatman no. 1 filter paper was then used to filter the supernatant, adjusted to a volume of 50 ml, and kept in an amber vial for further analysis.

2.4. Preliminary Phytochemical Screening

Using standard procedures [43], the presence of alkaloids, flavonoids, phenolics, tannins, steroids, triterpenoids, saponins, glycosides, carboxylic acids, anthraquinones, and essential oil was assessed. Briefly, to check for the presence of alkaloids, Wagner’s reagent was used. Triterpenoids were checked by using Salkowski’s reaction. The presence of flavonoids was confirmed by the lead acetate test showing yellow precipitate. Acetic anhydride test was carried out to check the presence of steroids. The presence of tannins was checked by adding 10% of NaOH and shaking well for an emulsion formation. Saponins were checked by foam test, where plant extract was mixed with water and shaken vigorously to see persistent foam for 10 min. Glycosides were checked using an aqueous NaOH test from the methanol extracts. Borntrager’s test determined the presence of anthraquinones. An effervescence test using a sodium bicarbonate solution was used to check carboxylic acid’s presence. To check for the presence of volatile oils, fluorescence tests were conducted. Details of the screening tests are shown in Table 1. The phytochemicals present in the methanol extracts of the leaves and flowers of D. angustifolia were compared.

2.5. Evaluation of Antioxidant Activity

We used the DPPH radical assay because it is easily available, has better radical scavenging potential, and is a commonly used free radical to evaluate the antioxidant potential of plant extracts [44, 45]. Modified protocol for the effect of free-radical scavenging on the DPPH assay from Banothu et al. [46] was followed. More briefly, to 0.25 mL of sample solution, 0.75 mL of DPPH was added and the reaction was left in the dark for 30 minutes. As a control, 0.25 mL of methanol and 0.75 mL of DPPH solution were combined. The standard utilized was ascorbic acid. In a 50 mL brown volumetric flask, 1.9716 mg of DPPH was dissolved in methanol to prepare DPPH solution and then adjusted to nearly 1.000 absorbance. A standard solution of ascorbic acid was prepared at a 1000 ppm concentration by dissolving 0.10 g in 100 ml of methanol for the calibration curve. From this stock solution, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, and 500 mg/L concentrations were prepared. The flower sample extract (0.1 g/mL) was diluted with methanol using 5, 10, 25, 50, 100, 150, and 200 dilution factors. The absorbance was measured using a JASCO V-770 spectrophotometer (Jasco, USA) at 517 nm with a 1 mm path length in a rectangular cell holder (500 μL cuvette). The percentage of inhibition is used to measure radical scavenging activity. The following formula was used to determine the DPPH radical scavenging capacity:

IC₅₀ values were computed from the relation log (sample) vs absorbance (normalized) using graph pad prism 8 software as suggested for better IC₅₀ estimation [47].

2.6. Total Phenol and Total Flavonoid Content Determination

The total phenol content (TPC) of the flower sample extract was determined by the Folin–Ciocalteu colorimetric method as described by McDonald et al. [48] with some modifications. More briefly, 0.2 mL of 2% Na₂CO₃ was added to the mixture after 0.4 mL of the extract and 0.4 mL of Folin–Ciocalteu reagent (10x diluted) were combined. Various concentrations of sample extracts were checked, and the one with dilution factor 5 was used for the determination as the absorbance was between 0.1 to 1 (within Beer’s Law) following consistent color changes. For control, a reagent without the methanol extract was used. The V-770 UV-Vis spectrophotometer (Jasco, USA) was used to measure the absorbance at 765 nm in triplicate after the mixture was incubated at room temperature for 35 minutes. The standard gallic acid was prepared for calibration at 1000 ppm and was serially diluted and 6.25, 12.5, 25, 50, 100, 150, 200, 250, and 500 mg/L standard solutions were prepared. Equation (2) was used to calculate TPC as the milligrams of gallic acid equivalent (GAE) per gram of extract (dry weight).where C is the concentration obtained from the calibration curve in mg/L, V is the final volume of the sample in L, m is the mass of the sample powder taken for extraction, and DF is the dilution factor.

Total flavonoid content (TFC) was determined using Chang et al.’s aluminum chloride colorimetric method [49] with slight modifications. More briefly, the mixture of 0.3 mL extract and 0.3 mL of 2% AlCl₃, 0.3 mL of 1% NaNO₂, and 0.3 mL of 5% NaOH were mixed and incubated at room temperature for a total of 30 min. Sample extracts were prepared in a similar way as for the TPC determination mentioned above. Methanol was used as a control. Absorbance was recorded in triplicate at λ 314 nm using a V-770 UV-Vis spectrophotometer (Jasco, USA). A standard stock solution of quercetin (1000 ppm) (mg/L) was prepared. The calibration standards were also prepared similarly. TFC was calculated as milligrams of quercetin equivalent (QE) per gram of the flower extract (dry weight).

2.7. HPLC Analysis

Stock solutions for standards (1 mg/mL) of both phenolic acids and flavonoids were prepared by dissolving an appropriate amount in methanol. Calibration standard solutions at 6 concentrations ranging from 2.5 to50 mg/mL were prepared and obtained by appropriate dilutions from the stock solutions in the selected mobile phase. The selected mobile phase was a binary isocratic elution consisting of (A) methanol and (B) acidified (1% acetic acid) ultra-pure water (60/40, v/v). The 10 μL injection volume was used at a flow rate of 0.8 mL/min.

Ultrahigh-performance liquid chromatography coupled with a diode array detector (Ultimate 3000 UHPLC-DAD, Thermo Scientific Dionex, USA) was used to separate the analytes via chromatography. The UHPLC system was equipped with a pump (model: LPG-3400SD), autosampler (model: WPS-3000TSL), and temperature-controlled column compartment (model: TCC-3100). Monitoring and quantitation were performed at 254 nm, 272 nm, 360 nm, and 372 nm. Chromeleon (c) Dionex version 7.2.4.8179 was used for instrument control and data acquisition. Chromatographic separation was performed on reverse phase column (Acclaim (TM) 102 with Fortis 5 μm, C18 (column dimension: 4.6 × 250 mm) Thermo Scientific Technologies, USA) operated at 30°C. Using these conditions, flavonoids and phenolic acid with external standards were separated within 25 min per sample. By comparing the retention times of the various compounds to standards, the individual compounds were identified and quantified. The amounts of the flavonoids and phenolic acids in the methanol extract of D. angustifolia flower were calculated using equation (2) [50].

2.8. PASS (Prediction of Activity Spectra for Substances) Test

PASS prediction for the flavonoids, phenolic acids, and reference antioxidant (ascorbic acid) was performed using the PASS online web server (http://www.pharmaexpert.ru/passonline). The PASS prediction uses canonical smiles to determine the probability of being active (Pa) and probability of being inactive (Pi) values. Pa and Pi values indicate a compound’s biological activity. The biological activities selected were those activities related in one way or another to antioxidant activity [51]. Interpretation of the result as recommended by Langunin et al. [39] and Maharani et al. [52] was as follows:(i)If Pa > 0.7, the compound is very active and is likely to display the aforementioned activity in trials conducted in a wet lab.(ii)If 0.5 < Pa < 0.7 is likely to exhibit the activity and there is a lower chance than in the first case, then the compound will demonstrate the activity in a wet lab experiment.(iii)If Pa < 0.5, the compound is unlikely to show the respective activity in the wet lab.

The 2.0 version of PASS online was used to perform the PASS test [53]. First, SMILES were retrieved for the candidate’s compounds from PubChem (http://pubchem.ncbi.nlm.nih.gov), then the MOL or SDF file for the compounds was given as input to the PASS software and activity prediction was performed (get prediction). Before conducting lab testing, it was crucial to confirm the results of the PASS biological activity test. The probability activity score would display the findings and indicate the likelihood of success if lab tests were conducted.

3. Results

3.1. Phytochemical Screening

The preliminary phytochemical screening revealed that the methanol extracts of the flower and the leaves were almost comparable qualitatively for the tested secondary metabolites. The glycoside, saponin, triterpenoid, and anthraquinone tests indicated more concentration in the leaves than in the flowers. Alkaloids and tannins were not detected in the flower extract (Table 2).

3.2. Antioxidant Activity Evaluation

The IC₅₀ values and % radical scavenging activity are almost identical for the leaf and flower extracts.

3.3. Total Phenol and Total Flavonoid Determination

Equation of calibration curve for TPC determination was y = 0.0023x − 0.0693, where R² = 0.9981, and for TFC determination was y = 0.002x + 0.0399, where R² = 0.9971.

3.4. HPLC Analysis

The number of components identified by HPLC analysis was 15 using a polar solvent system (Figure 1). Among these 6 were determined by using an external standard method on UHPLC-DAD (Table 3). The concentration (mg/100 g) was computed using V = 0.050 L and m = 5.0046 g on the equation given above in the method section.

Among the investigated flavonoids, myricetin took the lead with a concentration of 219.35 mg/100 g. Quercetin was 20x less than myricetin. The concentrations of rutin and gallic acid were found to be less than 10 mg/100 g. The concentration of syringic acid was 4x larger than chlorogenic acid. The data for kaempferol could not appear on the chromatogram and data table for unknown reasons after initially being tracked in the standard mixture. This might be due to precipitation and/or degradation of kaempferol at high temperatures (analysis temperature at 35°C). The rutin and gallic acid are not shown on the chromatogram due to the smaller amounts compared to the others.

3.5. PASS Prediction of Biological Activity

Biological activities used to describe the mechanism of antioxidant activity are considered in the PASS test. Other activities such as antimutagenic, cardioprotection, anticarcinogenic, chemopreventive, proliferative disease treatment, antibacterial, antiprotozoal, antifungal, anti-inflammatory, and antiviral activities were also considered. The PASS results (Supplement 1) are summarized for the compounds considered in Table 4.

4. Discussion

From the preliminary phytochemical screening test result (shown in Table 2), the leaves and flowers of D. angustifolia were similar in the constituency of the major phytochemical groups. Moreover, the results obtained for the leaf extract are in close agreement with the literature report [51]. Generally, the screening study indicated that flavonoids and phenolic compounds are the major constituents of the leaf and flower extracts of D. angustifolia [54].

The radical scavenging activity of both the leaves and flowers of D. angustifolia was assessed using the DPPH method. The percentage of inhibition used to measure the DPPH radical scavenging activity using the formula is as follows: percentage effect (E %) = (Abs_control − Abs_sample) × 100/Abs_control. As one can see from the structures of the flavonoids and phenolic acids, the presence of a more significant number of hydroxyl groups can result in higher antioxidant activity. As shown in Tables 5–7 and Figure 2, there is no significant difference in the antioxidant activity and TPC and TFC values between the leaves and flower parts of D. angustifolia.

(a)

(b)

Mobile phase selection for HPLC–DAD method for both extract samples was one of the big challenges to get better separation as the polarity of phenolic acids and flavonoids are closer. To come up with a solution for such problems, we have considered the advantage of the gradient method [55] for HPLC analysis. However, we successfully used isocratic elution for our flower sample. As shown on the chromatogram (Figure 1), the peak shape and separation were good for the selected polar solvent system. Methanol extract of D. angustifolia leaves was not showing separate peaks for the identified polyphenols in the case of the flower. Additional methods like hyphenation with mass spectroscopy should be considered for further component identifications.

The flavonoids (flavanols) considered in our study are sourced similarly following closely related biosynthesis in most cases. They are even common in dietary sources [56]. Flavonoids occur in most plant parts, specifically photosynthesizing plant cells as major coloring components of flowering plants [41]. For flowers, the matrix’s complexity may be lower compared to the leaves where chlorophyll and related components are more concentrated.

Investigations of phenolic acids and flavonoids among the phytochemicals from natural products are usually conducted using HPLC analysis with a DAD detector [57]. Such analysis has been attempted by Mizzi et al. (2020) [55]. Due to the complexity of the matrix analysis, it is not simple to do such an investigation for medicinal plants. Alam et al. [58] and Thomas et al. [59] mention an HPLC method using a gradient of acetonitrile and methanol to estimate some of these phenolics in Moringa oleifera. As the retention times of these compounds are closer to each other, making simultaneous measurements of both the groups was not possible in our case. Less than 1 mg/100 g of concentration was reported for rutin, myricetin, and gallic acid for D. viscosa flower [60]. Tong et al. reported gallic acid and quercetin 19.68 and 5.95 mg/100 g, respectively, from the flower of the same Dodonaea species [61]. In this study, HPLC analysis results are 5 to 220 mg/100 g for both flavonoids and phenolic acids. Myricetin was nearly 220 mg/100 g dry weight basis. Because of the few attempts on the synonym Dodonaea species, it is impossible to compare the results from this current investigation.

Biological activities predicted by PASS include main pharmacological effects, mechanisms of action, specific toxicities, interactions with antitargets, metabolic actions, influence in gene expression, and action on transporters [38, 53]. Biological activities related to antioxidant activity were selected for discussion from the activity spectrum of the individual flavonoids and phenolic acids under consideration. The Pi values were not significant for decision-making of inactivity for most of the compounds considered. On average, each of the considered flavonoids was predicted to have 90 pharmacological activities with Pa value greater than 0.700, which was doubled that of the standard reference compound. The gallic acid and syringic acid predicted pharmacological activities were 375 and 220, respectively. As a result, the flower of D. angustifolia can be considered a potential pharmacological agent [62], which is consistent with the therapeutic use of the plant’s leaves.

The antioxidant and other related activities of the four flavonoids, quercetin, rutin, myricetin, and kaempferol, have been predicted. Similarly, three phenolic acids, namely, gallic acid, syringic acid, and chlorogenic acid have been predicted using PASS. The PASS results were compared with ascorbic acid as a reference standard (Table 4). The Pa values for the flavonoids and chlorogenic acid were greater than 0.700. This indicates that the antioxidant activities of these compounds are very likely to be positive if attempted via wet lab experiments. These results agreed with the DPPH assay results of the extracts. The Pa value for ascorbic acid, a standard reference for in vitro antioxidant activity, was comparable with the flavonoids, more specifically myricetin and rutin.

Activities such as free radical scavengers, peroxidase inhibitors, membrane integrity agonists, dioxygenase inhibiter, and NADPH oxidase inhibitors refer to the mechanisms of action for the antioxidant activity by the respective compounds [51]. The Pa values of these activities were also greater than 0.700, indicating all to be among the possible mechanisms of action for the antioxidant activities of the compounds considered in this investigation. Considering the activation of internal antioxidants with Pa values less than 0.300, the polyphenols’ role in the activation of internal antioxidant enzymes was not expected.

The Pa values related to the antioxidant activity such as antimutagenic, cardioprotective, anticarcinogenic, chemopreventive, and proliferative disease treatment were also greater than 0.700. Except for minor inconsistencies in the treatment of proliferative illnesses, the compounds under consideration were also most likely projected to be similarly potent. Other activities such as antibacterials, antiprotozoal, antifungal, anti-inflammatory, and antiviral activities were predicted to show activities less likely in the wet lab experiments. Here, the probability will be less than the first case as Pa values are between 0.500 and 0.700, with few irregularities among the compounds and the activities. As flavonoids display varied cellular effects, they affect the overall process of carcinogenesis by varied mechanisms [63].

Phenolic acids: syringic acid, chlorogenic acid, gallic acid, and flavonoids: quercetin, myricetin, rutin, and kaempferol are common in food substances, including alcoholic drinks and fruits [56]. Phenolic compounds are known to impart beneficial properties such as antimicrobial, preservatives, antioxidants [55], and other varied physiological properties [64]. Antioxidant properties dictate so many pharmacological activities such as cardioprotective, anticarcinogenic, gastroprotective, anti-inflammation, and antimicrobial effects in the human body, and hence they are considered nutraceuticals [65]. As an example, rutin and quercetin showed the gastroprotective effects due to their antioxidant properties [66]. This is also shown on the biological activity spectrum for PASS online prediction. The mechanisms of such pharmacological activities include enzyme inhibition, disruption of cell membranes, blocking viral attachments and cell penetration, and activating the host cell’s self-defense mechanism [67].

5. Conclusions

D. angustifolia flower is a rich source of phytochemicals, which are extractable and can be checked for further biological activities depending on the phytochemical screening. A preliminary phytochemical study, the DPPH radical scavenging activity, and TPC, and TFC determinations all confirmed that the flowers and leaves of D. angustifolia are nearly similar in terms of phytoconstituency. From HPLC analysis, phenolic compounds identified clearly in the methanol extract of D. angustifolia flower include flavonoids: quercetin, myricetin, rutin, and phenolic acids: chlorogenic acid, syringic acid, and gallic acid. PASS biological activity prediction results show the stronger antioxidant activity of the identified flavonoids. Future work will emphasize the isolation and characterization of active principles responsible for bioactivity.

Data Availability

The data used to support this study are available in this article and the supplementary materials.

Conflicts of Interest

The authors declare that they have no conflicts of interest.

Acknowledgments

The authors acknowledge Dr. Belete Adefrse for his help in providing standards and Industrial Chemistry Department, Addis Ababa Science and Technology University and Woldia University for the opportunity to do this investigation.

Supplementary Materials

Supplementary file 1. PASS Data. (Supplementary Materials)

References

F. Beshah, Y. Hunde, M. Getachew, R. K. Bachheti, A. Husen, and A. Bachheti, “Ethnopharmacological, phytochemistry and other potential applications of Dodonaea genus: a comprehensive review,” Current Research in Biotechnology, vol. 2, pp. 103–119, 2020.
View at: Publisher Site | Google Scholar
E. Dagne, “Natural Database for Africa (NDA) Verion 2.0,” Addis Ababa, Ethiopia, 2011, http://alnapnetwork.com/NDA.aspx.
View at: Google Scholar
R. Fichtl and A. Adi, Honeybee flora of Ethiopia, Margraf Verlag, Weikersheim, Germany, 1994.
T. T. Jima and M. Megersa, “Ethnobotanical study of medicinal plants used to treat human diseases in berbere district, bale zone of oromia regional state, south East Ethiopia,” Evidence-based Complementary and Alternative Medicine, vol. 2018, Article ID 8602945, 16 pages, 2018.
View at: Publisher Site | Google Scholar
A. Belayneh, Z. Asfaw, S. Demissew, and N. F. Bussa, “Medicinal plants potential and use by pastoral and agro-pastoral communities in Erer Valley of Babile Wereda, Eastern Ethiopia,” Journal of Ethnobiology and Ethnomedicine, vol. 8, no. 1, pp. 42–11, 2012.
View at: Publisher Site | Google Scholar
G. Chekole, Z. Asfaw, and E. Kelbessa, “Ethnobotanical study of medicinal plants in the environs of Tara-gedam and Amba remnant forests of Libo Kemkem District, northwest Ethiopia,” Journal of Ethnobiology and Ethnomedicine, vol. 11, no. 1, p. 4, 2015.
View at: Publisher Site | Google Scholar
S. Zerabruk and G. Yirga, “Traditional knowledge of medicinal plants in Gindeberet district, Western Ethiopia,” South African Journal of Botany, vol. 78, pp. 165–169, 2012.
View at: Publisher Site | Google Scholar
G. Chekole, “Ethnobotanical study of medicinal plants used against human ailments in Gubalafto District, Northern Ethiopia,” Journal of Ethnobiology and Ethnomedicine, vol. 13, no. 1, p. 55, 2017.
View at: Publisher Site | Google Scholar
B. Desta, “Ethiopian traditional herbal drugs. Part I: studies on the toxicity and therapeutic activity of local taenicidal medications,” Journal of Ethnopharmacology, vol. 45, no. 1, pp. 27–33, 1995.
View at: Publisher Site | Google Scholar
S. Suleman and T. Alemu, “A survey on utilization of ethnomedicinal plants in nekemte town, East wellega (oromia), Ethiopia,” Journal of Herbs, Spices, and Medicinal Plants, vol. 18, no. 1, pp. 34–57, 2012.
View at: Publisher Site | Google Scholar
G. Seyoum and G. Zerihun, “An ethnobotanical study of medicinal plants in Debre Libanos Wereda, Central Ethiopia,” African Journal of Plant Science, vol. 8, no. 7, pp. 366–379, 2014.
View at: Publisher Site | Google Scholar
A. Belayneh and N. F. Bussa, “Ethnomedicinal plants used to treat human ailments in the prehistoric place of Harla and Dengego valleys, eastern Ethiopia,” Journal of Ethnobiology and Ethnomedicine, vol. 10, no. 1, pp. 18–17, 2014.
View at: Publisher Site | Google Scholar
A. Teklay, B. Abera, and M. Giday, “An ethnobotanical study of medicinal plants used in Kilte Awulaelo District, Tigray Region of Ethiopia,” Journal of Ethnobiology and Ethnomedicine, vol. 9, no. 1, pp. 65–23, 2013.
View at: Publisher Site | Google Scholar
S. Asnake, T. Teklehaymanot, A. Hymete, B. Erko, and M. Giday, “Survey of medicinal plants used to treat malaria by sidama people of boricha district, sidama zone, south region of Ethiopia,” Evidence-based Complementary and Alternative Medicine, vol. 2016, Article ID 9690164, 9 pages, 2016.
View at: Publisher Site | Google Scholar
T. T. Ayele, “A review on traditionally used medicinal plants/herbs for cancer therapy in Ethiopia: current status, challenge, and future perspectives,” Organic Chemistry: Current Research, vol. 7, no. 2, pp. 1–8, 2018.
View at: Publisher Site | Google Scholar
A. Enyew, Z. Asfaw, E. Kelbessa, and R. Nagappan, “Ethnobotanical study of traditional medicinal plants in and around Fiche District, Central Ethiopia,” Current Research Journal of Biological Sciences, vol. 6, no. 4, pp. 154–167, 2014.
View at: Publisher Site | Google Scholar
A. Hassan-abdallah, A. Merito, S. Hassan et al., “Medicinal plants and their uses by the people in the Region of Randa, Djibouti,” Journal of Ethnopharmacology, vol. 148, no. 2, pp. 701–713, 2013.
View at: Publisher Site | Google Scholar
T. Birhanu, D. Abera, and E. Ejeta, “Ethnobotanical study of medicinal plants in selected horro gudurru woredas, western Ethiopia,” Journal of Biology, Agriculture and Healthcare, vol. 5, no. 1, pp. 83–94, 2015.
View at: Google Scholar
G. L. Alemayehu, “Ethnobotanical Study on Medicinal Plants Used by Indigenous Local Communities in Minjar-Shenkora Wereda, North Shewa Zone of Amhara Region, Ethiopia,” Journal of Medicinal Plants Studies, vol. 3, no. 6, pp. 1–11, 2010.
View at: Google Scholar
T. Teklehaymanot, “Ethnobotanical study of knowledge and medicinal plants use by the people in Dek Island in Ethiopia,” Journal of Ethnopharmacology, vol. 124, no. 1, pp. 69–78, 2009.
View at: Publisher Site | Google Scholar
Y. Melaku, T. Worku, Y. Tadesse et al., “Antiplasmodial compounds from leaves of Dodonaea angustifolia,” Current Bioactive Compounds, vol. 13, no. 3, pp. 268–273, 2017.
View at: Publisher Site | Google Scholar
L. K. Omosa, B. Amugune, B. Ndunda et al., “Antimicrobial flavonoids and diterpenoids from Dodonaea angustifolia,” South African Journal of Botany, vol. 91, pp. 58–62, 2014.
View at: Publisher Site | Google Scholar
L. K. Omosa, J. O. Midiwo, S. Derese, A. Yenesew, M. G. Peter, and M. Heydenreich, “Neo -Clerodane diterpenoids from the leaf exudate of Dodonaea angustifolia,” Phytochemistry Letters, vol. 3, no. 4, pp. 217–220, 2010.
View at: Publisher Site | Google Scholar
R. Naidoo, M. Patel, Z. Gulube, and I. Fenyvesi, “Inhibitory activity of Dodonaea viscosa var. angustifolia extract against Streptococcus mutans and its biofilm,” Journal of Ethnopharmacology, vol. 144, no. 1, pp. 171–174, 2012.
View at: Publisher Site | Google Scholar
B. Mengiste, Y. Hagos, and F. Moges, “Invitro antibacterial screening of extracts from selected Ethiopian medicinal plants,” Momona Ethiop J Sci, vol. 6, no. 1, pp. 102–110, 2014.
View at: Google Scholar
T. Ngabaza, M. M. Johnson, S. Moeno, and M. Patel, “Identification of 5,6,8-Trihydroxy-7-methoxy-2-(4-methoxyphenyl)-4H-chromen-4-one with antimicrobial activity from Dodonaea viscosa var. angustifolia,” South African Journal of Botany, vol. 112, pp. 48–53, 2017.
View at: Publisher Site | Google Scholar
G. J. Amabeoku, P. Eagles, G. Scott, I. Mayeng, and E. Springfield, “Analgesic and antipyretic effects of Dodonaea angustifolia and Salvia africana-lutea,” Journal of Ethnopharmacology, vol. 75, no. 2-3, pp. 117–124, 2001.
View at: Publisher Site | Google Scholar
J. Tauchen, I. Doskocil, C. Caffi et al., “In vitro antioxidant and anti-proliferative activity of Ethiopian medicinal plant extracts,” Industrial Crops and Products, vol. 74, pp. 671–679, 2015.
View at: Publisher Site | Google Scholar
K. Teshome, T. Gebre-mariam, K. Asres, and E. Engidawork, “Toxicity studies on dermal application of plant extract of Dodonaea viscosa used in Ethiopian traditional medicine,” Phytotherapy Research, vol. 24, no. 1, pp. 60–69, 2010.
View at: Publisher Site | Google Scholar
K. Asres, F. Bucar, T. Kartnig, M. Witvrouw, C. Pannecouque, and E. De Clercq, “Antiviral activity against human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) of ethnobotanically selected Ethiopian medicinal plants,” Phytotherapy Research, vol. 15, pp. 62–69, 2001.
View at: Publisher Site | Google Scholar
T. Deressa, Y. Mekonnen, and A. Animut, “Vivo anti- malarial activities of Clerodendrum myricoides, Dodonea angustifolia and Aloe debrana against Plasmodium berghei,” The Ethiopian Journal of Health Development, vol. 24, no. 1, pp. 1–5, 2010.
View at: Publisher Site | Google Scholar
C. N. Muthaura, J. Keriko, C. Mutai et al., “Antiplasmodial potential of traditional phytotherapy of some remedies used in treatment of malaria in Meru – tharaka Nithi County of Kenya,” Journal of Ethnopharmacology, vol. 175, pp. 315–323, 2015.
View at: Publisher Site | Google Scholar
W. Amelo, P. Nagpal, and E. Makonnen, “Antiplasmodial activity of solvent fractions of methanolic root extract of Dodonaea angustifolia in Plasmodium berghei infected mice,” BMC Complementary and Alternative Medicine, vol. 14, pp. 462–467, 2014.
View at: Publisher Site | Google Scholar
B. Mengiste, E. Makonnen, and K. Urga, “In vivo antimalarial activity of Dodonaea angustifolia seed extracts against Plasmodium berghei in mice model,” Momona Ethiopian Journal of Science, vol. 4, no. 1, pp. 47–63, 2012.
View at: Publisher Site | Google Scholar
G. Mengistu, H. Hoste, M. Karonen, J. P. Salminen, W. H. Hendriks, and W. Pellikaan, “The in vitro anthelmintic properties of browse plant species against Haemonchus contortus is determined by the polyphenol content and composition,” Veterinary Parasitology, vol. 237, pp. 110–116, 2017.
View at: Publisher Site | Google Scholar
Z. K. S. Mcotshana, L. J. McGaw, D. Kemboi et al., “Cytotoxicity and antimicrobial activity of isolated compounds from Monsonia angustifolia and Dodonaea angustifolia,” Journal of Ethnopharmacology, vol. 301, Article ID 115170, 2023.
View at: Publisher Site | Google Scholar
S. Esakkimuthu, S. Mutheeswaran, P. Elankani, P. Pandikumar, and S. Ignacimuthu, “Quantitative analysis of medicinal plants used to treat musculoskeletal ailments by non-institutionally trained siddha practitioners of Virudhunagar district, Tamil Nadu, India,” Journal of Ayurveda and Integrative Medicine, vol. 12, no. 1, pp. 58–64, 2021.
View at: Publisher Site | Google Scholar
V. V. Poroikov, D. A. Filimonov, W. D. Ihlenfeldt et al., “PASS biological activity spectrum predictions in the enhanced open NCI Database Browser,” Journal of Chemical Information and Computer Sciences, vol. 43, no. 1, pp. 228–236, 2003.
View at: Publisher Site | Google Scholar
A. Lagunin, A. Stepanchikova, D. Filimonov, and V. Poroikov, “PASS: prediction of activity spectra for biologically active substances,” Bioinformatics, vol. 16, no. 8, pp. 747-748, 2000.
View at: Publisher Site | Google Scholar
A. Mishra, S. Kumar, and A. K. Pandey, “Scientific Validation of the Medicinal Efficacy of Tinospora Cordifolia,” Science World Journal, vol. 2013, Article ID 292934, 8 pages, 2013.
View at: Publisher Site | Google Scholar
C. S. Tiwari and N. Husain, “Biological activities and role of flavonoids in human health - a review,” Indian Journal of Scientific Research, vol. 12, no. 2, pp. 193–196, 2017.
View at: Google Scholar
F. Zakaria, J.-K. Tan, S. M. Mohd Faudzi, M. B. Abdul Rahman, and S. E. Ashari, “Ultrasound-assisted extraction conditions optimisation using response surface methodology from Mitragyna speciosa (Korth.) Havil leaves,” Ultrasonics Sonochemistry, vol. 81, Article ID 105851, 2021.
View at: Publisher Site | Google Scholar
J. R. Shaikh and M. Patil, “Qualitative tests for preliminary phytochemical screening: an overview,” International Journal of Chemical Studies, vol. 8, no. 2, pp. 603–608, 2020.
View at: Publisher Site | Google Scholar
D. M. Kasote, G. K. Jayaprakasha, and B. S. Patil, “Leaf disc assays for rapid measurement of antioxidant activity,” Scientific Reports, vol. 9, no. 1, pp. 1884–1910, 2019.
View at: Publisher Site | Google Scholar
R. Salazar, M. E. Pozos, P. Cordero, J. Perez, M. C. Salinas, and N. Waksman, “Determination of the antioxidant activity of plants from northeast Mexico,” Pharmaceutical Biology, vol. 46, no. 3, pp. 166–170, 2008.
View at: Publisher Site | Google Scholar
V. Banothu, C. Neelagiri, U. Adepally, J. Lingam, and K. Bommareddy, “Phytochemical screening and evaluation of in vitro antioxidant and antimicrobial activities of the indigenous medicinal plant Albizia odoratissima,” Pharmaceutical Biology, vol. 55, no. 1, pp. 1155–1161, 2017.
View at: Publisher Site | Google Scholar
Z. Chen, R. Bertin, and G. Froldi, “EC₅₀ estimation of antioxidant activity in DPPH assay using several statistical programs,” Food Chemistry, vol. 138, no. 1, pp. 414–420, 2013.
View at: Publisher Site | Google Scholar
S. McDonald, P. D. Prenzler, M. Antolovich, and K. Robards, “Phenolic content and antioxidant activity of olive extracts,” Food Chemistry, vol. 73, no. 1, pp. 73–84, 2001.
View at: Publisher Site | Google Scholar
C. C. Chang, M. H. Yang, H. M. Wen, and J. C. Chern, “Estimation of total flavonoid content in propolis by two complementary colometric methods,” Journal of Food and Drug Analysis, vol. 10, no. 3, pp. 178–182, 2020.
View at: Publisher Site | Google Scholar
E. C. Oliveira, E. I. Muller, F. Abad, J. Dallarosa, and C. Adriano, “Internal standard versus external standard calibration: an uncertainty case study of a liquid chromatography analysis,” Química Nova, vol. 33, no. 4, pp. 984–987, 2010, http://scholar.google.com/scholar?hl=en&btnG=Search&q=intitle:Quim.+Nova,#0%5Cnhttp://www.scielo.br/pdf/qn/v33n4/41.pdf.
View at: Publisher Site | Google Scholar
J. M. Lü, P. H. Lin, Q. Yao, and C. Chen, “Chemical and molecular mechanisms of antioxidants: experimental approaches and model systems,” Journal of Cellular and Molecular Medicine, vol. 14, no. 4, pp. 840–860, 2010.
View at: Publisher Site | Google Scholar
M. G. Maharani, S. R. Lestari, and B. Lukiati, “Molecular docking studies flavonoid (Quercetin, Isoquercetin, and Kaempferol) of single bulb garlic (Allium sativum) to inhibit lanosterol synthase as antihypercholesterol therapeutic strategies,” AIP Conference Proceedings, vol. 2231, p. 2020, 2020.
View at: Publisher Site | Google Scholar
D. A. Filimonov, A. A. Lagunin, T. A. Gloriozova et al., “Prediction of the biological activity spectra of organic compounds using the pass online web resource,” Chemistry of Heterocyclic Compounds, vol. 50, no. 3, pp. 444–457, 2014.
View at: Publisher Site | Google Scholar
S. O. Anode, T. Abraha, and S. Araya, “Phytochemical analysis of Dodonaea angustifolia plant extracts,” International Journal of Herbal Medicine, vol. 6, no. 5, pp. 37–42, 2018.
View at: Google Scholar
L. Mizzi, C. Chatzitzika, R. Gatt, and V. Valdramidis, “HPLC analysis of phenolic compounds and flavonoids with overlapping peaks,” Food Technology and Biotechnology, vol. 58, no. 1, pp. 1–12, 2020.
View at: Publisher Site | Google Scholar
A. P. B. Gollucke, D. A. Ribeiro, and O. J. Aguiar, “Polyphenols as supplements in foods and beverages: recent methods, benefits, and risks,” Polyphenols in Human Health and Disease, vol. 1, pp. 71–77, 2014.
View at: Publisher Site | Google Scholar
H. Kelebek, M. Jourdes, S. Selli, and P. L. Teissedre, “Comparative evaluation of the phenolic content and antioxidant capacity of sun-dried raisins,” Journal of the Science of Food and Agriculture, vol. 93, no. 12, pp. 2963–2972, 2013.
View at: Publisher Site | Google Scholar
P. Alam, S. F. Elkholy, S. A. Mahfouz, P. Alam, and M. A. Sharaf-Eldin, “HPLC based estimation and extraction of rutin, quercetin and gallic acid in Moringa oleifera plants grown in Saudi Arabia,” Journal of Chemical and Pharmaceutical Research, vol. 8, no. 8, pp. 1243–1246, 2016, https://www.jocpr.com/.
View at: Google Scholar
A. Thomas, A. Kanakdhar, A. Shirsat, S. Deshkar, and L. Kothapalli, “A high performance thin layer chromatographic method using a design of experiment approach for estimation of phytochemicals in extracts of Moringa oleifera leaves,” Turkish Journal of Pharmaceutical Sciences, vol. 17, no. 2, pp. 148–158, 2020.
View at: Publisher Site | Google Scholar
M. N. Malik, I. Haq, H. Fatima et al., “Bioprospecting Dodonaea viscosa Jacq.: a traditional medicinal plant for antioxidant, cytotoxic, antidiabetic and antimicrobial potential,” Arabian Journal of Chemistry, vol. 15, no. 3, Article ID 103688, 2022.
View at: Publisher Site | Google Scholar
Z. W. Tong, H. Gul, M. Awais et al., “Determination of in vivo biological activities of Dodonaea viscosa flowers against CCl₄ toxicity in albino mice with bioactive compound detection,” Scientific Reports, vol. 11, no. 1, Article ID 13336, 2021.
View at: Publisher Site | Google Scholar
V. Poroikov, D. Filimonov, A. Lagunin, and A. Stepanchikova, “PASS: prediction of biological activity spectra for substances,” Bioinformatics, vol. 16, no. 8, pp. 747-748, 2000.
View at: Publisher Site | Google Scholar
S. Ramos, “Effects of dietary flavonoids on apoptotic pathways related to cancer chemoprevention,” The Journal of Nutritional Biochemistry, vol. 18, no. 7, pp. 427–442, 2007.
View at: Publisher Site | Google Scholar
S. Bajkacz, I. Baranowska, B. Buszewski, B. Kowalski, and M. Ligor, “Determination of flavonoids and phenolic acids in plant materials using SLE-SPE-UHPLC-MS/MS method,” Food Analytical Methods, vol. 11, no. 12, pp. 3563–3575, 2018.
View at: Publisher Site | Google Scholar
A. Tapas, D. Sakarkar, and R. Kakde, “Flavonoids as nutraceuticals: a review,” Tropical Journal of Pharmaceutical Research, vol. 7, no. 3, pp. 1089–1099, 2008.
View at: Publisher Site | Google Scholar
M. Morsy and A. El-Sheikh, “Prevention of gastric ulcers,” in Peptic Ulcer Disease, J. Chai, Ed., InTech, Rang-Du-Fliers, France, 2011.
View at: Publisher Site | Google Scholar
M. Friedman, “Overview of antibacterial, antitoxin, antiviral, and antifungal activities of tea flavonoids and teas,” Molecular Nutrition and Food Research, vol. 51, no. 1, pp. 116–134, 2007.
View at: Publisher Site | Google Scholar

Copyright

Copyright © 2023 Fekade Beshah Tessema et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PDF Download Citation

Download other formats

Order printed copies

Views

994

Downloads

560

Citations