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Research Detail

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Sium Ahmed
Department of Biotechnology and Genetic Engineering, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Faisal Bin Rahman
Department of Biotechnology and Genetic Engineering, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Shawon Ahmed
Department of Biotechnology and Genetic Engineering, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Abdullah Mohammad Shohael*
Department of Biotechnology and Genetic Engineering, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Tomato (Solanum lycopersicum L.) is a widely cultivated vegetable crop worldwide and its consumption is increasing day by day. The only edible part of tomato plant is its fruit. Therefore, the residual tomato plant parts are considered as waste after fruit collection despite some portion of it is used as livestock feed and in the production of fertilizer. Moreover, due to the consumption issue, much of the research is focused on tomato fruit rather than other plant parts. Bioactive compounds can be present in any part of the plant and can be isolated and recovered. Therefore, tomato plant may be useful as a source of bioactive compounds. Our study focuses on the phytochemical constituents and bioactive compounds that are present in different parts of the tomato plant that were being less studied before. Qualitative phytochemical tests were performed to identify the presence of different bioactive compounds. Saponins, tannins, glycosides, phenols, alkaloids, flavonoids and resins are present in significant amount. Total phenol, total flavonoids, total tannin, and total protein content were measured quantitatively. Leaf samples showed higher concentrations of bioactive compounds than roots and stem. Highest total phenol content (66.43 mg/g), total flavonoids content (28.00 mg/g), total tannin content (7.36 mg/g) and total protein content (26.55 mg/g) was found from the leaf water extract.  DPPH scavenging assay was performed to find the antioxidant potential and positive results were found. Thin layer chromatography (TLC) was also employed to check the constituents. Our study found several spots for different samples and their retention factors (Rf) were evaluated.  

  Antioxidants, Phytochemicals, Tomato, TLC.
  Plant Biotechnology and Genetic Engineering Laboratory, Department of Biotechnology and Genetic Engineering, Jahangirnagar University, Dhaka-1342, Bangladesh
  
  
  Development of Host and Medicinal Plants
  Medicinal Plants

The aim of this study was to investigate the presence of phytochemicals in the plant extracts obtained from roots, stem, and leaves and their quantification. The research also armed to find out the antioxidant potential as well as the Thin Layer Chromatographic profiling to evaluate the phytochemical composition.

Chemicals and reagents: All chemicals and reagents used in this study were of analytical grade. Aluminum chloride, (+)-Catechin, Potassium iodide, DPPH (2,2diphenyl-1-picrylhydrazyl), Gallic acid were purchased from Sigma Aldrich, USA. Glacial acetic acid was purchased from Scharlau, Spain. Tannic acid was purchased from Loba Chemie, India. Bradford reagent was purchased from Biobasic Canada. Ethanol, Methanol, Fehling A and B, Sodium hydroxide, Sodium nitrite were bought from Merck, Germany. Folin-Ciocalteu reagent was purchased from SRL, India. Sodium carbonate, Chloroform, Iodine were bought from Alfa Aesar, UK. Hexane was bought from Daejung chemicals, South Korea. Ethyl acetate was bought from Duksun pure chemicals, South Korea.   Plant material: The experiments were conducted at the Plant Biotechnology and Genetic Engineering Laboratory, Department of Biotechnology and Genetic Engineering, Jahangirnagar University, Dhaka-1342, Bangladesh (23°53’14” N 90°15’56” E). The tomato plants (Solanum lycopersicum L.) of variety Ratan were used in this study. The seeds were collected from the local market and planted on the seedbed. 30 days old healthy seedlings were planted in the experimental field located on departmental garden. The plots were maintained according to the standard tomato production management system. 60 days old plants were collected for the preparation of samples.  Preparation of Extract: The plant materials (leaf, stem, and root) were shade dried and powdered in a mechanical grinder. The powdered material (10g) were taken in conical flasks with ethanol, methanol, and water as solvent and kept them in a shaker for 3 days at room temperature. The extracts were filtered through Whatman no. 1 filter paper and filtered organic solvents were concentrated using an evaporator, and finally, stock solutions of extracts (10 mg/mL) were prepared. Qualitative Phytochemical Analysis: The freshly prepared crude extracts were qualitatively tested for the presence of secondary metabolites especially Carbohydrates, saponins, tannins, glycosides, phenols, coumarins, alkaloids, flavonoids, resins and terpenoids through the following methods. Carbohydrates: 1 mL of different extracts were taken into test tubes to which equal volume of Fehling’s A and Fehling’s B were added. The tubes were heated at 65°C in a water bath for 10-15 min. Red brick precipitate indicates the presence of carbohydrates (Fehling, 1849). Saponins: 2 mL of distilled water added to 1 mL of extracts and then shaken for 5 min. The presence of 1cm foam for 10 min indicates the presence of saponin (Kumar et al., 2009).  Tannins: .5 mL of extracts were mixed with 1 mL of distilled water, and then 4-5 drops of 1% ferric chloride were added. Blue color and the greenish black color indicate the presence of gallic tannin and cathecholic tannin respectively (Ahamed et al., 2017).  Glycosides: 1mL of extracts were mixed with 1ml of glacial acetic acid. Then 5-6 drops of 1% ferric chloride solution was added. Brown color ring produced at the top indicate glycoside’s presence (Ahamed et al., 2017). Phenols: 1ml of ethanol was added to 1ml of extracts. Then 6-7 drops of 1% ferric chloride solution was added to each tube. Formation of the green, blue, purple color indicates the presence of Phenol (Soloway et al., 1952). Coumarins: 1 ml of extract was added with 1.5 ml of 10% NaOH. The chemical reaction will produce a yellow color and indicate the presence of coumarin (Ugochukwu et al., 2013). Alkaloids: In 1 ml of extract, 4-6 drops of Wagner’s reagents [1.27gm of Iodine and 2gm of potassium iodide in 100ml of water] was added. The Radish brown precipitate indicates the presence of Alkaloid (Rizk, 1982). Flavonoids: 3-4 drops of 20% NaOH solution was added in 2ml of extract. The intense yellow color formed and become colorless when 4-5 drops of diluted HCl was added. This indicates the presence of Flavonoids (Ugochukwu et al., 2013). Resins: 1 mL of extracts were taken into test tubes. Few drops of acetic anhydride were added to each tube. Finally, 1 mL of concentrated H2SO4 was added carefully. The presence of yellow to orange color confirmed the presence of resins (Iqbal et al., 2015). Terpenoids: 5 mL of extracts were taken into test tubes and mixed with 2 mL of chloroform. Then concentrated H2SO4 was added to each tube. The appearance of reddish-brown coloration at the interface indicated the presence of terpenoids (Sheel et al., 2014).  Determination of total phenolic content: The total phenolics content of the extracts was estimated according to the method described previously (Shohael et al., 2006). 0.5 mL aliquots of extracts and standards of different concentrations were mixed with 2.5 mL tenfold diluted Folin-Ciocalteu reagent and 2.5 mL 7.5% sodium carbonate. Then they are kept for 30 min at 25°C. The absorbance was measured at 765 nm in a UV-visible spectrophotometer (T60 UV-Visible Spectrophotometer, PG Instruments Ltd., United Kingdom). The concentration of phenol was determined gallic acid as standard.  Total phenolic content was expressed as milligrams of Gallic Acid equivalent (mg GAE)/g extract. Determination of total flavonoid content: Total flavonoid content was determined following the method of Sakanaka et al., (2005) modified by Shohael et al., (2006). Briefly, 0.25 mL of plant extract or (+)-Catechin standard solution was mixed with 1.25 mL of distilled water, followed by addition of 75 μL of a 5% sodium nitrite solution. After 6 min, 0.150 ml of a 10% aluminum chloride solution was added, and the mixture was allowed to stand for a further 5 min and then 0.5 ml of 1 M sodium hydroxide was added. The mixture was brought to 2.5 ml with distilled water and mixed well. The absorbance was measured immediately at 510 nm in a UV-visible spectrophotometer (T60 UV-Visible Spectrophotometer, PG Instruments Ltd., United Kingdom).  The concentration of flavonoids was determined using (+)-Catechin as standard. The concentration of flavonoids was expressed as milligram Catechin equivalent (mg CE)/g extract. Determination of total tannin content: The tannins were determined using the Folinciocalteu reagent as described previously (Amorim et al., 2008). Briefly, 0.1 ml of the sample extract is added with 7.5 ml of distilled water.  Then 0.5 mL of Folin-ciocalteu reagent and 1 mL of 35% sodium carbonate solution were added. Total volume was adjusted to 10 mL by adding distilled water. The mixture was incubated at room temperature for 30 min and the absorbance was measured at 725 nm in a UV-visible spectrophotometer (T60 UV-Visible Spectrophotometer, PG Instruments Ltd., United Kingdom). Blank was prepared with water instead of the sample. A set of standard solutions of tannic acid is read against a blank. The concentration of tannin was determined using tannic acid as standard. The concentration of tannin was expressed as milligram Tannic Acid equivalent (mg TAE)/g extract. Determination of total protein content: Total proteins were estimated according to the method described by Bradford (Bradford, 1976). 0.1 ml of different extract were taken into test tubes and then 0.9 ml Bradford reagent was added and incubated for 2 min. The absorbance was measured at 595 nm in a UV-visible spectrophotometer (T60 UV-Visible Spectrophotometer, PG Instruments Ltd., United Kingdom). The concentration of protein was determined using Bovine Serum Albumin (BSA) as standard. The concentration of protein was expressed as milligram Bovine Serum Albumin equivalent (mg BSAE)/g extract. Statistical analysis: Significant differences among mean values were compared by Tukey's honestly significant difference (HSD) test at a level of significance of p ≤ 0.05. All data were displayed as the mean ± standard error of the mean at least three independent biological replications. The statistical analysis was performed using Statistical Package for Social Science software (SPSS, version 16.0, IBM Corporation, NY).

  Jahangirnagar University J. Biol. Sci. 7(2): 65-77, 2018 (December)
  
Funding Source:
1.   Budget:  
  

Therefore, the result shown here is not a constant indicator of the compounds present. However, more intense research should be employed to evaluate the potential of the tomato plant as a source of bioactive compounds that may be useful in different aspect of pharmaceutical, industrial and biotechnological applications.

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