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Table of Contents
ORIGINAL ARTICLE
Year : 2021  |  Volume : 5  |  Issue : 4  |  Page : 77-83

Chemical contents in relation to the antimicrobial efficacies of Solanum spp. sect. petota


1 Department of Pure and Applied Zoology, Federal University of Agriculture, Abeokuta, Nigeria
2 Department of Biological Sciences, Fourah Bay College, University of Sierra Leone, Freetown, Sierra Leone
3 Department of Microbiology, University of Ilorin, Ilorin, Nigeria
4 Department of Science Laboratory Technology, Microbiology Unit, Kwara State Polytechnic, Ilorin, Nigeria
5 Department of Public Health and Policy Management, College of Medicine, University of Ibadan, Ibadan, Nigeria

Date of Submission03-Dec-2021
Date of Acceptance03-Jan-2022
Date of Web Publication20-Apr-2022

Correspondence Address:
Dr. Adeyinka O Adepoju
Department of Biological Sciences, Fourah Bay College, University of Sierra Leone, Freetown
Sierra Leone
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/mtsp.mtsp_12_21

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  Abstract 


Background: Both Solanum lycopersicum and S. Pimpinellifolium possess much nutritional and therapeutic usefulness. Aim: This study is aimed at investigating the phytochemical, chemical and antimicrobial contents of foliar extracts of both plants. Methodology: The analyses were carried out using standard methods; the organic contents of the extracts were determined by GC-MS before the extracts were tested for antibacterial and antifungal potentials using the disc diffusion method. Results: The biochemical analysis revealed the presence of crude fibre, crude fat, crude carbohydrate, moisture content, total ash, dry matter and crude protein. Various mineral elements such as Manganese, Zinc, Copper, Sodium, Potassium, Magnesium, Calcium, Iron and Phosphorus were detected in the extracts, even as GC-MS revealed 6 different organic compounds belonging to two groups of chemicals (ester and alkanol). The methanolic extract of both plants showed significant antibacterial and antifungal activities on some of the test organisms at a dose range of 5 and 25mg/ml. Cardiac glycosides was observed to be the highest constituent of the phytochemicals detected and it has been reported to inhibit sodium and potassium pump which in turn cause an increase in the amount of calcium ions and are thus useful in the treatment of heart failure and cardiac arrhythmia. Conclusion: This study concluded that the leaf extracts of both plants investigated are highly nutritive, containing appreciable amounts of nutrients which are required in human and animal diet. Recommendation: An advocacy for a higher consumption of these tomatoes as well as their use in the formulation of antimicrobial agents and drugs is highly recommended.

Keywords: Antimicrobials, GCMS, minerals, petota, phytochemicals, solanum


How to cite this article:
Aladesida AA, Adepoju AO, Adesola-Famade B, Ajiboye TO, Fagbolu TA. Chemical contents in relation to the antimicrobial efficacies of Solanum spp. sect. petota. Matrix Sci Pharma 2021;5:77-83

How to cite this URL:
Aladesida AA, Adepoju AO, Adesola-Famade B, Ajiboye TO, Fagbolu TA. Chemical contents in relation to the antimicrobial efficacies of Solanum spp. sect. petota. Matrix Sci Pharma [serial online] 2021 [cited 2022 Jun 26];5:77-83. Available from: https://www.matrixscipharma.org/text.asp?2021/5/4/77/343598




  Introduction Top


Since antiquity, plants have been employed not just as a source of food, clothing, and shelter but also for therapeutic use in traditional medicine. Over 1340 plants with defined antimicrobial activities are said to exist and over 30,000 antimicrobial compounds have been isolated from plants.[1],[2] Plants which are used for therapeutic purposes are referred to as medicinal plants. Different parts of medicinal plants such as leaves, roots, rhizomes, stems, barks, flowers, fruits, grains, or seeds have been used to prevent and treat ailments; ranging from respiratory illnesses, urinary tract infection, gastrointestinal system illness as well as skin infections.[3]

Medicinal plants are said to be the richest source of antimicrobial agents; a single plant could have many antimicrobial properties and could therefore be used for the treatment of several ailments. In recent years, herbal medicines have attracted enormous attention as significant alternatives to commercial and synthetic drugs, even among literates in urban regions. This could be as a result of some adverse effects caused by certain antibiotics (e.g., hypersensitive reaction to penicillin and ototoxicity by aminoglycosides); increasing cost of antibiotics, which in turn affects maintenance of health; and increase in antibiotic resistance by various bacteria e.g., Methicillin-resistant Staphylococcus aureus, Klebsiella pneumoniae.[4],[5]

The two species under study; Solanum lycopersicum (tomato) and Solanum pimpinellifolium (currant tomato, a small-sized tomato-like fruit), belong to a taxonomic hierarchical group named Petota section; order Solanales and family Solanaceae.[6],[7] S. pimpinellifolium are the smallest edible tomato, they grow on intermediate vines. Like S. lycopersicum, the plants may get up to 8 feet tall and require staking to keep fruit exposed to light and off the ground. The leaves of the plant are very tiny and delicate with a more acrid odor than other tomato variants). A tomato plant has compound leaves usually 10–25 cm long, made up of leaflets which are about 5–9 on petioles. The leaflets are 8 cm long with serrated margin and have a weak stem which often sprawls over the ground.[7],[8]

S. lycopersicum has a widespread distribution. It is highly and widely consumed either in its raw form or in its processed form as paste, puree, juice, or ketchup.[9],[10] It is a rich source of vitamins, minerals, dietary fibers, essential amino acids, and carotenoids; lycopene and flavonoids.[11],[12] The leaves of tomato have a higher content of antimicrobial metabolites such as chlorogenic acid, α-tomatine, α-terpinene, sabinene, caffeic acid, β-phellandrene, vanillic acid, dehydrotomatine than its fruits.[13] Aside from the many benefits, the fruit of these plants offers, the leaves are also an exceptional source of lycopene, a naturally occurring pigment, and an antioxidant. It is known to help prevent, fight and repair cell damage in the human body, support healthy eyesight, cardiovascular health, and a lot more.

Even though the classification of some members of the family Solanaceae (to which these plants belong) has been reviewed,[14] it is essential to also look at some relationship between their chemical content and their beneficial properties. In this study, the antibiotic capability of the extracts of S. lycopersicum and S. pimpinellifolium against selected clinically important microorganisms as well as their phytochemical and chemical contents was investigated.


  Materials and Methods Top


Collection of plant materials

S. lycopersicum and S. pimpinellifolium leaf samples were collected into sterile polythene bags from the Botanical Gardens, Ladoke Akintola University of Technology (LAUTECH) research plot where they have been previously grown. The leaves were identified at Lautech Herbarium, Biology Laboratory Complex, LAUTECH, Nigeria. The harvested leaves were cleaned using potable water and air-dried for 4 weeks in the general biology laboratory complex, LAUTECH, Nigeria. The dried leaves of the plants were thoroughly crushed into fine powder using an electric blender and preserved in an airtight container in readiness for the next set of analysis.

Extraction of active ingredients from the plant materials

Extractions were carried out by soaking 20 g of each of the powders in 200 ml of absolute methanol in well-labeled clean conical flasks and corked. After 7 days of extraction, the soaked leaf powders were removed and the decanted solutions were concentrated on a rotary evaporator after they have been filtered using Whatman no 1 filter paper. The dried extracts were kept in a refrigerator in readiness for various analyses. Enough quantity of powder was kept for various analyses for which they were needed. The extracts were stored in sterile covered sample bottles to keep them from evaporating and then getting more concentrated before use.

Growth and maintenance of test organisms for antimicrobial studies

Bacterial culture of different strains, which included Pseudomonas aeruginosa, Pseudomonas putida, Bacillus cereus, Bacillus subtilis, Escherichia coli, S. aureus, K. pneumoniae (maintained at 37°C), fungal culture of Aspergillus flavus, Candida albicans, Aspergillus niger (maintained at 25°C) and yeast culture of Saccharomyces cerevisiae (maintained at 25°C) were obtained from the microbial gene bank of Pure and Applied Biology Department, LAUTECH, Ogbomoso, Nigeria.[15]

Preparation of inoculum

To prepare bacteria inoculum, 5 ml of Nutrient broth was pipetted into each test tube, corked, and autoclaved for 15 min at 121°C; test tubes were then arranged on a slide rack and labeled. Each test tube was inoculated with 2 loop-full of the test bacteria and mixed very well. The mixtures were transferred into the incubator for 2 h at 37°C after which visible growth of the organisms was observed. Fungi inoculum was prepared by mixing 0.2 g of Yeast extract with 1 g of sucrose and 100 ml of water inside a beaker. Five ml of this was pipetted in each test tube, covered with a cork and autoclaved for 15 min at 121°C to sterilize. The media was allowed to cool and two loopful of test fungi were inoculated in each of the bottles and incubated. After 2 h, there was a notable growth of fungi.[15]

Antimicrobial assay of plant extract

The antimicrobial and antifungal activities were evaluated using the disc diffusion method. Petri plates were prepared with 20 ml sterile nutrient agar for bacteria and Potato dextrose agar for fungi and the bottom of the plates were divided into segments 5, 25, 50, 100, and 250 respectively with the control at the center, with the names of the organisms boldly written on each plate. The test cultures were swabbed on top of the solidified media and the perforated filter paper was put into the extracts of varied concentration (0.25 mg/ml, 0.1 mg/ml, 0.005 mg/ml, and 0.0025 mg/ml) was put into the marked area that matches and the disc with only ethanol. The bacterial plate was incubated in the incubator at 37°C for 24 h and fungi at 28°C for 48 h.[16]

Antimicrobial assay of broad-spectrum antibiotics

The test organisms were swabbed on the surface of the solidified sensitivity agar using sterile swab sticks to obtain a lawn swab. Two types of antibiotic discs were used depending on the Gram staining property of the test organism under observation, that is, Gram-positive or Gram-negative. The plates were slightly opened and an appropriate antibiotic disc was placed inside. The plates are not turned upside down but they are incubated at 37°C for 24 h. After incubation, the zones of inhibition around each antibiotic disc were measured.

Proximate analysis

Ten grams of each of the powders were exhaustively processed for various parameters according to the methods described by the Association of Physical Analytical Chemists.[17] By the use of weight difference, ash and moisture contents were obtained. The fiber content was estimated from the loss in weight of crucible and its content on drying. Carbohydrate was determined by subtracting the sum of the percentages of moisture, crude protein, ash, and fats from 100. The determination of nitrogen value (precursor of protein of a substance) was by micro Kjeldahl method which involves digestion, distillation, and finally titration of the sample. The nitrogen value was then converted to protein by multiplying it by a factor of 6.25. Crude lipids content was determined by the use of Soxhlet type of direct solvent extraction method using petroleum ether boiling at 50°C. The nitrogen free extracts were calculated indirectly by difference as the sum of crude protein, fiber, fats and ash subtracted from 100. All the results of proximate analysis were expressed in percentages.

Phytochemical analysis

Phytochemical analysis for tannins, phenol, flavonoids, terpenoids, saponins, phlobatannin, cardiac glycosides and alkaloids were carried out according to known and standard methods. Tannins and phlobatannins were estimated using the Folin-Denis spectrophotometric method. Saponin content was determined using the method of Liener and modified by Hudson and El-Difrawi. Flavonoids were determined by ethyl acetate extraction and gravimetric measurement, Alkaloids by the alkaline precipitation, and gravimetric method by Harborne and Cardiac glycosides by Keller-Killian's test. For Phenols, 2 ml of distilled water was added to 1 mg of plant sample followed by a few drops of 10% aqueous ferric chloride solution. The formation of blue or green color indicated the presence of phenols.[18],[19],[20]

Mineral content analysis

Minerals were determined after the dried powdered samples were first digested with nitric acid and perchloric acid and the filtered aliquot was used for the determination of Na, K, Ca, Mg, P, Fe, Cu, Mn, and Zn content. K and Na were determined by the flame photometric method. Fe, Cu, Zn, Mn, Ca, Mg and Zn were determined by the atomic absorption spectrophotometric method described by James (1995) and the Association of Official Analytical Chemists (2010).[21],[22]

Gas chromatography-mass spectrophotometry analysis

The gas chromatography-mass spectrophotometry (GC–MS) analysis was conducted by an Agilent 6890 GC system with Agilent 6971 inert mass selective detector (Agilent Tech. Palo Alto, California, USA). Separations were carried out on capillary column, HP5 5% phenyl methyl siloxane nonpolar column (30 000 mm × 0.32 mm diameter, with a layer thickness of 0.25 mm stationary phase. The oven temperature was set at a temperature of 80°C and maintained for 5 min. Then, the oven temperature was raised 15°C/min to a temperature of 300°C and maintained for 2 min. The carrier gas used was helium ultrapure (99.999%). The temperature of injector, interface, and ion source were 310, 280, and 230°C, respectively. The injection volume was 1 mL, using a model with a 1:10 split inlet, with gas flow rate in the column at 1.3 mL/min.[16]

Statistical analysis

All experiments were carried out in triplicate and results were expressed as Mean ± standard deviation these data were subjected to analysis of variance using SPSS for Windows, Version 16.0. Chicago, SPSS Inc. was obtained at P < 0.05.


  Results Top


Antimicrobial activities of methanolic extract of Solanum lycopersicum and S. pimpinellifolium on some test organisms

The antimicrobial effects of the leaves extracts are shown as zones of inhibition in millimetres (mm) on some selected microorganisms; 7 bacteria, 3 fungi, and 1 yeast, which includes, P. aeruginosa, P. putida, B. cereus, K. pneumoniae, B. subtilis, E. coli, S. aureus, A. niger, A. flavus, C. albicans, and S. cerevisiae are presented in [Figure 1]a and [Figure 1]b. The antimicrobial activity of the leaf extract of S. Pimpinellifolium was found to be the highest at 5 mg/ml against E. coli which produced an inhibition zone of 12 mm [Figure 1]a while that of S. lycopersicum is highest at 5 and 25 mg/ml against C. albicans with a zone of inhibition of 12 mm [Figure 1]b.
Figure 1: (a) Effect of methanolic extract of Solanum Pimpinellifolium extract on the test organisms. (b) Effect of methanolic extract of S. Solanum lycopersicum extract on the test organisms. Keys: P.A – Pseudomonas aeruginosa, P.P – Pseudomonas putida, B.C – Bacillus cereus, K.P – Klebsiella pneumoniae, B.S – Bacillus subtilis, E.C – Escherichia coli, S.A – Staphylococcus aureus, A.N – Aspergillus niger, A.F – Aspergillus flavus, C.A –Candida albicans and S.C – Saccharomyces cerevisiae, 25% – 25% conc., 5% – 5% conc., GEN/AUG – Gentamicin/Augumentin, C - Control

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The foliar organic compounds in the species of Solanum through gas chromatography-mass spectroscopy

[Table 1] shows the results of the GC-MS analysis conducted on the leaves of the species of Solanum studied. A total of 6 organic compounds were detected and they belong to 2 different groups of chemicals namely, ester and alkanol. 5 of the organic compounds were esters while just one belonged to an alkanol [Table 1]. Hexadecanoic acid methyl ester (HEAME), 9-octadecenoic acid methyl ester (E-) and Phytol were identified as the chief chemical compounds ranging from 15.55% to 38.33% while the other three were present in minor quantities with peak areas ranging from 1.39% to 5.91%.
Table 1: The percentage peak area values of some organic compounds quantified in the leaves of the Solanum species studied

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Phytochemicals in the leaves of the species of Solanum studied

[Table 2] shows the presence of some metabolites in the leaf extract of the Solanum species studied. The methanolic extract of S. lycopersicum and S. pimpinellifolium leaves contained alkaloids, saponin, tannin, cardiac glycosides, flavonoids, phenols, phlobatannin and terpenoids.
Table 2: Mean quantities of some secondary metabolites (ppm) in the leaves of the Solanum species studied

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Saponins have also been reported in Senna alata and Cajanus cajan by Lawal et al.,[23] in Lophira lanceolata seeds by Lohlum et al.[24] and also in cultivars of Capsicum by Adepoju et al.[15] These phytochemicals have been associated with antimicrobial activities.[25],[26] Apart from their antimicrobial activities, the secondary metabolites observed in this study such as tannins are known to have astringent properties, useful in treatment of inflamed tissue and in prevention of cancer. Flavonoids, saponins and cardiac glycosides act as antiparasitic agents, antiviral agents, can inhibit tumor growth and are useful as antidepressant.[3],[27],[28] The content of tannin in the extract of S. lycopersicum and S. pimpinellifolium was not significantly different at P ≤ 0.05 while that of alkaloids, saponin, cardiac glycosides, flavonoids, phenols, phlobatannin, and terpenoids differed significantly at P ≤ 0.05. The mean quantities of the phytochemicals ranged from 0.12-0.92, with Cardiac glycosides having the highest constituent (0.92) and Phenols the lowest (0.12).

The biochemical composition of the leaves extracts of Solanum lycopersicum and Solanum pimpinellifolium

The percentage proximate composition of the methanolic extract of S. lycopersicum and S. pimpinellifolium leaves is presented in [Table 3]. The results of the proximate composition of the leaf extracts showed the presence of crude fiber, crude fat, crude carbohydrate, moisture content, total ash, dry matter, and crude protein. Moisture content and crude protein occurred in appreciable amounts.
Table 3: Percentage proximate composition of the species of Solanum studied

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The content of crude fiber and total ash were also high, this thereby suggesting the high nutritive value of these plant materials. The high total ash values are an indication of high mineral content. Of all the nutrients detected, the leaves were particularly high in moisture content (36%–37%) but low in dry matter (2.3%).

The composition of crude fiber, crude carbohydrate, moisture content, crude protein, and total ash is higher in S. lycopersicum than in S. pimpinellifolium. The result shows a significant difference at P ≤ 0.05 only in the composition of crude fibre, crude carbohydrate, moisture content and crude protein in the species of Solanum studied. The quantities of the proximate composition of the Solanum species studied can be listed from the highest to the lowest in this order–moisture content > protein > crude-fiber > total ash > crude fat > crude carbohydrate > dry matter.

Mineral element composition of the extracts of Solanum lycopersicum and Solanum pimpinellifolium

[Table 4] shows the mineral element composition of the species of Solanum plants studied.
Table 4: Mineral element composition (ppm) of the species of Solanum plants studied

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The result of the mineral composition of the leaf extracts showed the presence of Manganese, Zinc, Copper, Sodium, Potassium, Magnesium, Calcium, Iron, and Phosphorus. Potassium (27.4%–27.6%), Calcium (27.1%–27.3%) and Magnesium (13.1%–13.3%) were detected in appreciable quantities while the remaining six elements were in trace amounts. On the whole, the quantities of the mineral elements in the plants studied can be enumerated from the highest to the lowest as

Potassium > Calcium > Magnesium > Sodium>

Zinc > Copper > Phosphorus > Iron and Manganese.

Calcium is essential for growth and maintenance of bones, teeth, and muscles. Potassium is an important extracellular cation while Magnesium is a component of chlorophyll and is both required for chemical reactions in the body and intestinal absorption.[29],[30],[31],[32] The high levels and collective presence of these three elements are known to reduce hypertension and blood pressure. They are therefore essential in the prevention and treatment of hypertension.[17],[29]

Significance of organic, proximate, mineral, and phytochemical contents on antimicrobial efficacy of the extracts

In this study, five of the identified volatile compounds belong to the class ester and one to the class alkanol, and these have been reported to be pharmacologically active. HEAME is said to have potential antibacterial and antifungal activities as well as antioxidant properties.[33],[34] Crude fibers are necessary in diet for digestion and effective to eliminate the risks of coronary heart disease, hypertension, constipation and breast cancer. There is said to be a strong correlation between crude fibre and moisture content, as the fiber is easily digested and disintegrated which could be of high interest in human nutrition. Since both nutrients are high in these plants, they are therefore valuable sources of dietary fiber needed for human health.[35],[36] Also the high total ash values are an indication of high mineral content which was later observed in the study.

Minerals are required for normal growth, muscular activities, and skeletal development. The high levels and collective presence of Calcium, Potassium, and Magnesium are known to reduce hypertension and blood pressure. They are therefore essential in the prevention and treatment of hypertension.[17],[29] Copper, Iron, and Manganese although observed in trace amounts are still essential for cellular activity, oxygen transport, and intestinal absorption and chemical reaction in the body respectively.[15],[37]

The presence of alkaloids, saponins, tannin, phlobatannins, anthraquinones, cardiac glycosides, flavonoids, and phenol in these extracts may be responsible for the good antibacterial activity. Phenolic compounds are known to have antimicrobial effects.[38] Flavonoid derivatives were reported to be effective antimicrobial substances against different microorganisms. Their mode of action may be due to their ability to complex with extracellular and soluble proteins and bacterial cell wall. In addition to having antibacterial activity, flavonoids are also effective against viruses and parasites.[38]

In earlier studies, the antimicrobial, proximate, organic, mineral, and phytochemical analyses of the extract of the tomato plant have been carried out but very few have been done on the foliar extract of the plant.[7],[13],[26],[39] According to Kim et al.(2014), the tomato leaves include a higher content of antimicrobial metabolites than tomato fruits and also contain more phenolic compounds and alkaloids than other tomato parts. It is therefore important that further studies be carried out on the foliar extract of these plants.[39]

The significant antimicrobial activity of methanolic extracts of Solanum species might be due to the synergistic effects of compounds identified in the proximate, organic, mineral, and phytochemical analyses of the plant part. Something striking to note is that S. lycopersicum extract was found to be most active against C. albicans at 5 and 25% concentration with a zone of inhibition that suggests a better performance than the test drugs.

Based on this study, it can be observed that the leaves of the plant material (S. lycopersicum and S. Pimpinellifolium) are of high nutritive value, as they contain an appreciable amount of nutrients which are important requirements in human and animal diet. The high total ash composition is an indicator of a high mineral content which is evident in the result of the mineral element composition. The extracts contain several phytochemicals; alkaloids, saponins, cardiac glycosides, tannins, phlobatannin, phenol, flavonoids, and terpenoids. The presence of these detected phytochemicals is responsible for the antimicrobial activity of these species of Solanum studied as the test organisms were resistant to most of the broad-spectrum antibiotics investigated, while no resistivity was observed against the extracts of the plant materials used. Cardiac glycosides were observed to be the highest constituent of the phytochemicals detected and it has been reported to work by inhibiting sodium and potassium pump which in turn cause an increase in the amount of calcium ions and are thus useful in the treatment of heart failure and cardiac arrhythmia. A high composition of cardiac glycosides (which was discovered in the plants hereby studied) is relative to high calcium content.


  Conclusion Top


This study has proved that methanolic extract of the leaves of S. lycopersicum and S. pimpinellifolium possess microbe inhibitory properties, owing to their constituents and can therefore be employed in the production of broad-spectrum antimicrobial agents for use in medicine and pharmaceutical industries. This suggests that the plants are good alternative source of antimicrobial agents and show more promising antimicrobial potentials than some drugs. The authors wish to recommend that S. lycopersicum and S. pimpinellifolium should be further studied for their potentials towards utilization in drug production.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Tables

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



 

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