|Year : 2021 | Volume
| Issue : 3 | Page : 307-312
Antioxidant, antibacterial, and cytotoxic activities of different parts of Salsola vermiculata
Saeed Mollaei1, Jalaledin Ghanavi2, Poopak Farnia3, Parvin Abedi-Ghobadloo1, Ali Akbar Velayati2
1 Department of Chemistry, Phytochemical Laboratory, Faculty of Sciences, Azarbaijan Shahid Madani University, Tabriz, Iran
2 Mycobacteriology Research Centre, National Research Institute of Tuberculosis and Lung Disease (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran
3 Mycobacteriology Research Centre, National Research Institute of Tuberculosis and Lung Disease (NRITLD); Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
|Date of Submission||06-Jun-2021|
|Date of Acceptance||07-Aug-2021|
|Date of Web Publication||7-Sep-2021|
Dr. Jalaledin Ghanavi
Mycobacteriology Research Centre, National Research Institute of Tuberculosis and Lung Disease, Shahid Beheshti University of Medical Sciences, Tehran
Mycobacteriology Research Centre, National Research Institute of Tuberculosis and Lung Disease, Shahid Beheshti University of Medical Sciences, Tehran; Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran
Source of Support: None, Conflict of Interest: None
Background: Salsola vermiculata is an annual plant which is broadly distributed in southwest of Asia. Due to the lack of data regarding the biological properties of different parts of S. vermiculata, the present research focused on their biological activities. Methods: Different parts of plant were harvested, and phytochemical assay of S. vermiculata parts was performed to determine the presence of phenols, flavonoids, alkaloids, fixed oils, and volatile oils. Then, their antioxidant and cytotoxicity activities of the extracts and fractions were evaluated according to DPPH and MTT assay, respectively. Furthermore, the antimicrobial activities of the extracts and fractions were determined by the disk diffusion and minimum inhibitory concentration methods. Results: According to the results, the antioxidant activity and the phenolic contents in leaves and flowers crude extracts of S. vermiculata were high, and their ethyl acetate fractions had higher activities compared to other fractions. The cytotoxic study indicated that the seeds and roots crude extracts significantly exhibited cytotoxic activities with IC50 values of 33.88, 42.92 μg/mL, respectively, and only the fixed oil fractions had cytotoxic activity. Antibacterial activity showed that the fixed oil fraction of roots had the highest activity against Staphylococcus aureus, followed by fixed oil fraction of seeds and volatile oil fraction of the roots. Conclusions: Different parts and fractions of S. vermiculata contain various bioactive compounds with different biological activities, and the results of this study could provide useful information to guide the application of S. vermiculata parts in food and pharmaceutical fields.
Keywords: Antibacterial, antioxidant, cytotoxic, metabolite
|How to cite this article:|
Mollaei S, Ghanavi J, Farnia P, Abedi-Ghobadloo P, Velayati AA. Antioxidant, antibacterial, and cytotoxic activities of different parts of Salsola vermiculata. Biomed Biotechnol Res J 2021;5:307-12
|How to cite this URL:|
Mollaei S, Ghanavi J, Farnia P, Abedi-Ghobadloo P, Velayati AA. Antioxidant, antibacterial, and cytotoxic activities of different parts of Salsola vermiculata. Biomed Biotechnol Res J [serial online] 2021 [cited 2021 Nov 26];5:307-12. Available from: https://www.bmbtrj.org/text.asp?2021/5/3/307/325613
| Introduction|| |
Herbal and natural medicines play significant roles in the treatment of diseases and development of novel drugs. Therefore, the search for finding plants with specific biological activity would be an interesting field of knowledge. Salsola is a genus belonging to the family of Amaranthaceae in the major group of Angiosperms, which is native to Asia, Europe, and Africa. These plants are widespread across the hypersaline, semiarid, and arid areas.,, Many plants of this genus are used to cure skin diseases, human heart ailments, influenza, and cough. In addition, various species of the genus Salsola have displayed medicinal properties such as controlling the obesity, diabetes, and Alzheimer's disease; functioning as the anticancer, anti-inflammatory, antibacterial, antihypertensive, and antioxidant agents; central nervous system depressant activity; and being the cure for tape worm infestation.,,,,
Salsola vermiculata is an annual plant which is broadly distributed in southwest of Asia and belongs to Amaranthaceae family. This plant is used for the treatment of stomach disorders. Rasheed et al. investigated the metabolite profile of S. vermiculata and could identify several flavonoids, hydroxycinnamic acids, fatty acids, and alkaloids. Al-Tohamy et al. indicated high antioxidant and antimicrobial activities of S. vermiculata methanolic extract. Therefore, due to the lack of data regarding the biological properties of different parts of S. vermiculata, the present research focused on the biological activity of different parts of S. vermiculata.
| Methods|| |
The study was approved by Scientific Committee of NRITLD & Azarbaijan Shahid Madani University at 11.1.2017.
The S. vermiculata plant was collected from natural habitats in Azarshahr, Iran, during October 2017. The S. vermiculata parts were separated from each other and then dried at room temperature in darkness for 7 days and powdered. Furthermore, voucher specimen after identifying (ASMUH-10485) was deposited in the official Azarbaijan Shahid Madani University.
Fractionation of crude extract
One gram of the each part of plant was extracted 3 times with 10 mL of 80% methanol at room temperature using ultrasonic apparatus. After centrifugation, the supernatants were subjected to reduced pressure to obtain sticky residues. The crude methanolic extract was suspended in 10 mL of distillated water, which was subsequently extracted 3 times with n-hexane (1:l v/v) to separate none polar compounds from crude extraction. The n-hexane layer was distillated using Clevenger apparatus, and two fractions including volatile oil (fraction 1) and fixed oil (fraction 2) were obtained. Then, the aqueous phase was treated with acetic acid (1%) and partitioned with ethyl acetate (1:l v/v). The aqueous acid phase (fraction 3) and ethyl acetate phase (fraction 4) were separated and then were dried under nitrogen at room temperature and transferred to the vials.
Screening of phytochemicals
Phytochemical analyses of S. vermiculata parts (roots, seeds, leaves, and flowers) were performed to determine the presence of phenols, flavonoids, alkaloids, fixed oils, and volatile oils. To test the presence of phenols, ferric chloride solution (10% v/v) was added to the extract, and formation of blue or green color indicated the presence of phenols. Alkaline reagent test was used to confirm the presence of flavonoids. An aqueous solution of the extracts was treated with NH4OH solution followed by addition of a few drops of concentrated H2SO4. The appearance of a yellow color verified the presence of flavonoids. Dragendorff's test was utilized to determine the presence of alkaloids in the extracts. Dragendorff's reagent was added to a few milliliter of the extract. The appearance of a yellow precipitate confirmed the presence of alkaloids. The presence of anthocyanins was confirmed by addition of 2N HCl solution to the extract. The appearance of a pink-red color which turned purplish blue after addition of NH3 indicated the presence of anthocyanins. The presence of fixed oils in the extracts was checked based on pressing of a small quantity of extract between two papers. The oil stain on the paper showed the presence of fixed oils. Salkowski test was utilized to determine the presence of terpenoids in the extracts. The extract was mixed with chloroform and concentrated H2SO4. The appearance of a reddish-brown interface confirmed the presence of terpenoids.
Determination of antioxidant activity by DPPH assay
The antioxidant activity of the samples was investigated using our previous method with slight modification. Briefly, 100 μL of the sample was mixed with 100 μL of DPPH (0.01 mM). The obtained solution was incubated at 25°C for 30 min, and then, the absorbance was read at 517 nm.
The cytotoxicity activity of the S. vermiculata extracts was evaluated according to MTT assay. Briefly, human lung cancer cell line A549 was cultured in RPMI-1640 medium, and then, the cell solution (100 μL, 1 × 104 cells/mL) was added to 96-well plates. The culture media were treated with different concentrations of the extracts. After 72 h, 20 μL of MTT solution was added to each well and the mixture was incubated for 4 h. Finally, the MTT absorption was read at 570 nm, and the cell viability (%) was calculated.
The extracts of S. vermiculata organs were tasted separately Bacillus subtilis, Bacillus cereus, Bacillus pumilus, Staphylococcus aureus, Klebsiella pneumonia, Escherichia coli, and Staphylococcus epidermidis. The disk diffusion method using Mueller–Hinton agar plates and Sabouraud dextrose agar was used for the determination of antibacterial activity against bacteria. In addition, the broth microdilution assay was used for the determination of the minimum inhibitory concentration (MIC) values.
All instruments were performed in triplicate. The analysis was done by the SAS 9.2 using a completely randomized design (one-way analysis of variance), and the mean comparisons were determined by Tukey's test (P < 0.05).
| Results|| |
Phytochemical screening of crude methanolic extracts and their fractions of different parts of S. vermiculata indicated the presence of some important bioactive components, which are listed in [Table 1]. The crude extract showed high concentrations of terpenoids in roots, terpenoids and oils in seeds, flavonoids and terpenoids in leaves; phenols and flavonoids in flowers, moderate concentrations of alkaloids and oils in roots; phenols in leaves, low concentrations of phenols and flavonoids in roots, alkaloids, phenols and flavonoids in seeds, anthocyanins and oils in leaves; anthocyanins, terpenoids and oils in flowers; absence of anthocyanins in roots and seeds; and alkaloids in leaves and flowers. The analysis of volatile oil fractions indicated high concentrations of terpenoids in the roots and seeds; moderate concentrations of oils in the roots and seeds; low concentrations of oils and terpenoids in the leaves and flowers; and the absence of alkaloids, phenols, flavonoids, and anthocyanins in all parts. The analysis of fixed oil fractions indicated high concentrations of oils in the roots and seeds; terpenoids in leaves; moderate concentration of terpenoids in the roots and seeds; low concentrations of oils in leaves; terpenoids and oils in flowers; and absence of alkaloids, phenols, flavonoids, and anthocyanins in all parts. The ethyl acetate fraction exhibited high amounts of phenols and flavonoids in leaves and flowers; moderate amounts of anthocyanins in flowers; low amounts of phenols and flavonoids in roots and seeds; anthocyanins in leaves; the absence of alkaloids, anthocyanins, terpenoids, and oils in roots and seeds; and alkaloids, terpenoids, and oils in leaves and flowers. On the other hand, the aqueous acid fractions showed high concentrations of alkaloids in roots; low concentration of alkaloids, and anthocyanins in seeds and flowers, respectively; and the absence of phenols, flavonoids, anthocyanins, terpenoids, and oils in roots and seeds; alkaloids, phenols, anthocyanins, terpenoids, and oils in flowers; and all studied metabolites in leaves.
|Table 1: Phytochemical screening of crude methanolic extracts and their fractions of different parts of Salsola vermiculata|
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Results of antioxidant activity were obtained using the DPPH method and are shown in [Figure 1]. According to the results, IC50 values of the roots, seeds, leaves, and flowers crude extracts were 164.91 ± 3.91, 64.63 ± 3.73, 36.41 ± 1.44, and 37.29 ± 1.61 μg/mL, respectively. Furthermore, since the antioxidant activity and the levels of phenolic compounds in leaves and flowers crude extracts were high, the antioxidant activity of their fractions was also studied. Results indicated that ethyl acetate fractions had higher antioxidant activities compared to other fractions [Figure 1]b and [Figure 1]c.
|Figure 1: DPPH radical scavenging activity of (a) methanolic extracts of different parts of Salsola vermiculata; (b) different fractions of Salsola vermiculata leaves; (c) different fractions of Salsola vermiculata flowers|
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Moreover, total phenolic content of flowers in aqueous acid, ethyl acetate, fixed oil, and volatile oil fractions was found to be 2.3, 12.8, 1.3, and 1.5 (mg GAE/g), respectively, and in the leaves were 1.4, 9.4, 0.8, and 0.7 (mg GAE/g), respectively. The total flavonoid content of the fractions indicated that the ethyl acetate fractions of leaves and flowers had the highest amounts of flavonoid compounds (2.9 and 3.4 mg QE/g), while there is no flavonoid content in the fixed oil and volatile oil fractions of leaves and flowers.
The cytotoxic activity of the S. vermiculata parts against the A549 cell line was studied, and the results showed that the seed and root crude extracts significantly exhibited cytotoxic activities with IC50 values of 33.88, 42.92 μg/mL, respectively. In addition, cytotoxicity results revealed that other parts of plant had no cytotoxic properties (IC50 values were up to 1000 μg/mL) [Figure 2]a. The study on cytotoxic activity of the roots and seeds fractions indicated that only fixed oil fractions had cytotoxic activity [Figure 2]b.
|Figure 2: Cytotoxic activity of (a) methanolic extracts of different parts of Salsola vermiculata; (b) the fixed oil fractions of roots and seeds against A549 cell lines|
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The antibacterial activities of the crude extracts of various organs of S. were evaluated and the results are shown in [Table 2] and [Table 3]. The inhibition zones were between 10.29 ± 0.21 and 18.37 ± 0.31 mm. According to the results, all of the crude extracts illustrated moderate to high inhibitory activity, and the behavior of the extracts was different against the studied bacteria., Among the studied extracts, the most sensitive microorganism was S. aureus in the case of root and seed extracts, and MIC values were mostly <7.50 mg/mL [Table 2]. The leave and flower extracts indicated moderate antibacterial activity against all of the studied microorganisms, except for S. aureus with inhibition zone of 15.21 ± 0.27 mm, which showed higher antibacterial activity [Table 2].
|Table 2: In vitro antimicrobial activities of the Salsola vermiculata extracts (disk diffusion method) against various microorganisms|
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|Table 3: Minimum inhibitory concentration (minimum inhibitory concentration [mg/mL]) of the Salsola vermiculata extracts|
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Due to the most antibacterial activity of the roots and seeds extracts against S. aureus, their fractions were investigated against S. aureus bacteria [Table 4]. According to the results, the fixed oil fraction of roots had the highest activity against S. aureus, followed by fixed oil fraction of seeds and volatile oil fraction of the roots.
|Table 4: In vitro antimicrobial activities of the Salsola vermiculata fractions (minimum inhibitory concentration [mg/mL]) against Staphylococcus aureus|
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| Discussion|| |
The present research focused on the biological activities of different parts of S. vermiculata. According to phytochemical studies, S. vermiculata contained various metabolites such as terpenoids, oils, flavonoids, phenols, and alkaloids. Each phytochemical showed potency toward some biological action; for example, phenols and flavonoids play a role in antioxidant potential;,,, and antibacterial and cytotoxic activities were found with fatty acids.,,,
The phytochemical screening of the ethyl acetate fractions of the leaves and flowers indicated the presence of phenol and flavonoid compounds. In addition, the findings of total phenolic content indicated that these fractions are enrich in these fractions. So far, several phenolic compounds have been reported from different species of Salsola. Shehab and Abu-Gharbieh reported a number of phenolic compounds in methanolic extract of S. imbricata, categorized as flavonoids, phenolic acids, and simple phenolic compounds, while coumaric acid and quercetin were the main detected compounds. Boulaaba et al. analyzed the methanolic extracts of S. kali and concluded that this plant contains seven phenolic compounds. In another study, the aerial part of S. vermiculata was shown to be more enriched in rutin and kaempferol derivatives versus root samples. Previous researches resulted that there is a positive correlation between the phenolic and flavonoid levels and antioxidant activity., These compounds probably act as the reduction factor and react with radical compounds and change them into resistant components, which eventually result in neutralization of free radical chain., Consequently, thanks to unique properties of the phenolic compounds, the fractions possessing these compounds might have antioxidant activities. Therefore, the antioxidant activity of the ethyl acetate fractions of the leaves and flowers was investigated. It became clear that these fractions had the highest antioxidant activities compared to others. Finally, it can be concluded that the antioxidant activities could be related to the presence of phenolic compounds.
According to previous researches, fatty acids are known to have potential antibacterial agents., Palmitic acid was reported as antibacterial compound against Gram-negative and Gram-positive bacteria. Dilika et al. reported that oleic and linoleic acids had antibacterial activity only against the Gram-positive bacteria and were inactive against the Gram-negative bacteria. Skalicka-Woźniak et al. proposed that oleic and linoleic acids, the main compounds in Peucedanum species, were responsible for antibacterial activity. Furthermore, according to previous studies, the fatty acids play a crucial role in cell proliferation and apoptosis induction., Harada et al. indicated the cytotoxicity activity of palmitic acid against human leukemic cells, but this compound had no cytotoxicity effect on normal HDF cells. Moreover, palmitic acid induces apoptosis in MOLT-4 cell line. Fujiwara et al. indicated that linolenic acid suppresses cell growth of neuroblastoma cell lines. Moreover, Belury introduced linolenic acid as a chemotherapeutic agent. Consequently, thanks to unique properties of these fatty acids, the fractions possessing these compounds might be potentially applied as a promising source of cytotoxic and antibacterial agents. Therefore, the antibacterial and cytotoxic activities of the fixed oil fractions of roots and seeds were investigated. The results showed that the fixed oil fraction of roots had the highest activity against S. aureus, followed by fixed oil fraction of seeds. Moreover, the extracts of S. vermiculate were more sensitive to Gram-positive bacteria. Furthermore, the fixed oil fractions of the roots and seeds had cytotoxic activity. Hence, it can be concluded that the high cytotoxic and antibacterial activity of the fixed oil fraction could be related to the presence of palmitic, oleic, and linoleic acids.
| Conclusion|| |
In this investigation, the biological activities of different parts and fractions of S. vermiculata were studied. Phytochemical screening of different parts indicated the presence various metabolites (phenolic compounds, alkaloids, fatty acids, and volatile compounds). Studying the biological activities of the fractions of different parts of S. vermiculata indicated the highest antioxidant activity of ethyl acetate fraction of leaves, the highest cytotoxic activities of the fixed oil fractions of the roots and seeds, and the highest antibacterial activities of fixed oil fraction of roots. In conclusion, different parts and fractions of S. vermiculata contain various bioactive compounds with different biological activities, and the results of the present study could provide useful information to guide the application of S. vermiculata parts in food and pharmaceutical fields.
We gratefully acknowledge financial support from Azarbaijan Shahid Madani University and Shahid Beheshti University of Medical Sciences.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3], [Table 4]