Biotechnology Research and Innovation (2018) 2, 1 --- 8 http://www.journals.elsevier.com/biotechnology-research-and-innovation/ RESEARCH PAPER Phytochemical constituents of Dracaena mahatma leaves and their anti-bacterial, anti-oxidant and anti-inflammatory significance a a a a Saranya Shankar , Sugashini Settu , Gayathri Segaran , Ranjitha Dhevi V. Sundar , b,∗ Lokesh Ravi a School of Biosciences and Technology, VIT University, Vellore 632014, Tamil Nadu, India b Composite Interceptive Med-Science Laboratories Pvt Ltd., Bangalore 560099, Karnataka, India Received 19 March 2018; accepted 15 September 2018 Available online 28 September 2018 KEYWORDS Abstract The present study aims to analyze the antimicrobial, antioxidant and anti- Ornamental plant; inflammatory activity of Dracaena mahatma leaf extract. Qualitative phytochemical screening Dracaena mahatma; of leaf powder confirmed the presence of tannins, alkaloids, phenols, terpenoids, sterols, Antibacterial activity; triterpenes and anthraquinone glycosides. GC-MS analysis of methanolic extract suggested Gas the presence of 2 unknown compounds and 8 known phytochemicals, of which benzene-(1,6- chromatography-mass hexanediylidene)tetrakis constituted a maximum percentage of the crude extract. The total spectrometry; phenolic content of the crude extract was found to be 32.9 ± 0.002 GAE/g. Crude extract Antioxidant; demonstrated 90% antioxidant activity in DPPH assay at 100 ␮g/ml concentration and also Anti-inflammatory demonstrated significant antioxidant activity in FRAP assay at 100 ␮g/ml concentration. The crude extract did not show any antagonism against fungal pathogens Aspergillus niger and Aspergillus flavus. Crude extract demonstrated significant antibacterial activity against 5 stud- ied pathogen, with the highest antagonism against Escherichia coli with a zone of inhibition of 22 mm at 10 mg/well concentration. Introduction at moderate temperatures, humidity and moisture condi- tions. This plant can be grown well in pots as well as ground (Palanivelu, Kunjumon, Suresh, Nair, & Ramalingam, 2015). Dracaena mahatma is an ornamental plant which remains Aslam et al. reported that numerous steroidal saponins that colorful in bright light and in semi-shade. Dracaena leaf are produced by Dracaena draco exhibited cytostatic activ- remains attractive and colorful for a longer period of time ity on Leukemia HL 60 cells. The clotting process was greatly improved in mice by the extract obtained from the Dracaena ∗ cochinchinensis. The stem of D. draco contains a red resin Corresponding author. known as Dragon’s blood, which has been used as traditional E-mail: [email protected] (L. Ravi). https://doi.org/10.1016/j.biori.2018.09.002 2452-0721/© 2018 Sociedade Brasileira de Biotecnologia. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). 2 S. Shankar et al. sealed with parafilm and placed in a shaker for 48 h at 120 rpm. After 48 h the content was filtered using a Whatman filter paper No. 1. The methanol solvent was evaporated by using a rotary vacuum evaporator. The crude extract was concentrated and further studies have been carried out using this extract. The solvent extract was done once for every 15 g of sample with 200 ml methanol, in a single flask. Qualitative analysis of phytochemicals Using standard protocol, phytochemicals such as phe- nol, alkaloid, saponin, tannin sterol and triterpenes and anthraquinone glycoside were analyzed (Andriani et al., 2015; Sundar, Segaran, Shankar, Settu, & Ravi, 2017). Figure 1 D. mahatma plant sample used for this study. Water extract medicine as an ‘‘herbal remedy’’ (Aslam, Mujib, &, Sharma, To the 14 ml of distilled water, add 1 g of D. mahatma dried 2013). leaf powder and using mantle this content was boiled for Dendranthema grandiflorum is a hybrid ornamental 5 min. Then the content was filtered using Whatman No. 1 plant, mainly grown for flower farming and also used filter paper to obtain water extract in the production of insecticidal products. The bioactive Phenol test. To the 1 ml of the filtrate, add few drops of compounds isolated from this plant showed larvicidal activ- FeCl3, the appearance of dark green color indicates the pres- ity against Aedes aegypti larvae (Spindola et al., 2016). ence of phenol. Aslam et al. (2013) reported that several activities like Saponin test. In a test tube add 2 ml of distilled water antileishmanial, antimalarial, molluscicidal, fungicidal and along with the small amount of crude extract and shaken bacteriostatic activities are showed by Spirocanozole-A an vigorously. The presence of saponin can be confirmed by the active saponin compound extracted from Dracaena arborea formation of stable foam. and Dracaena mannini. Tannin test. Add few drops of 2% FeCl3 to the test tube con- Based on these literature surveys and based on the lack taining crude extract, the presence of tannin was confirmed of research articles on D. mahatma and due to the reported by the display of blue-green color. bioactivity of this genera of plants, this plant was chosen as the plant of interest for this analysis. This study ana- Acid extract lyzed the antibacterial, antioxidant and anti-inflammatory In the beaker, add 1 g of dried leaf powder and 5 ml of con- activity of D. mahatma leaves and also analyses the phy- centrated HCl, then the content was mixed gently and allow tochemical composition of the extract by the biochemical it to stand for 20 min. Using Whatman filter paper the filtrate reaction and GC-MS method (Fig. 1). was filtered. Flavonoid test. To the 2 ml of 2% sodium hydroxide solu- Scientific classification tion, add a small amount of filtrate, on the addition few drops of diluted acid, the yellow colored solution turned to Kingdom: Plantae colorless, this shows the presence of flavonoid. Clade: Angiosperms Clade: Monocots Alcohol extract Order: Asparagales Few milliliters of methanol was added to the leaf powder Family: Asparagaceae and allowed to stand for 30 min then the content was fil- Subfamily: Nolinoideae tered using Whatman filter paper. The filtrate was kept on Genus: Dracaena the mantel until the solution get evaporates. Finally, to Species: Mahatma resuspend, 3 ml of chloroform were added to it. Alkaloid test. In a filter paper, few drops of alcohol extract Materials and methods were deposited, and then over the filter paper Dragendroff’s reagent was splashed. The presence of alkaloid was con- firmed by the appearance of reddish brown color. Leaf crude extract preparation Sterol and triterpenes test. Few drops of concentrated H2SO4 and acetic anhydride were added to the sidewise of Fresh leaves of D. mahatma were collected in the sterile the test tube containing 2 ml of alcoholic extract. A positive bag from VIT University, Vellore, Tamil Nadu, India. The result can be confirmed by the appearance of reddish brown leaves were washed thoroughly with double distilled water color. to remove dust and foreign particles. Then the leaves were Anthraquinone glycoside test. A volume of 1 ml ammonia shade dried for 3 --- 4 days. Once it gets completely dried, the was mixed with extract then the mixture was shaken vigor- leaves were ground into a fine powder. ously, the positive result was indicated by the appearance of The leaf powder of about 10---15 g was dissolved in 200 ml green color in the bottom and reddish color in the aqueous of methanol in an Erlenmeyer flask. Then the flask was layer. Bioactivity of Dracaena mahatma 3 ◦ Determination of total phenol content (MTCC 3160), then it was incubated at 37 C for 24 h. Further, the bacterial culture was streaked over the surface of the Folin-Ciocalteu photometric method was carried out to Muller-Hinton agar plate. Using cork borer wells were made determine total phenol content of plant extract. A small on the plates and these wells were filled with 100 ␮l leaf amount of methanolic plant extract was oxidized with Folin- extract at different concentrations (25, 50 and 100 mg/ml). Ciocalteu reagent. Then the saturated sodium carbonate The crude extract was dissolved in 1% DMSO solution, to was added to neutralize the reaction. To dilute the solution obtain the different concentrations. Ampicillin disk was used 50 ml volume of distilled water was used. Then the sam- as positive control and these plates were kept for incuba- ◦ ples were incubated at room temperature for 90 min, the tion at 37 C for 24 h. After the incubation period, the zone absorbance was measured at 750 nm. Gallic acid was used of inhibition was measured (Abew, Sahile, & Moges, 2014). as a standard solution (Senguttuvan, Paulsamy, & Karthika, All experiments were performed in triplicates and average 2014). was considered as the final result. Antioxidant activity: DPPH radical scavenging assay Antifungal activity The DPPH radical scavenging assay is a standard procedure Antifungal activity of the methanolic crude extract was to determine the antioxidant activity of the leaf extract. assayed by agar well diffusion method. Fungal pathogens Aliquots of methanolic extract at different concentrations such as Aspergillus niger (MTCC 3323) and Aspergillus flavus (25, 50, 75, 100 mg/ml) were added to 2 ml of DPPH. (MTCC 2799) were cultured and maintained in PDA (Hi- Gallic acid was prepared at different concs (25, 50, 75, media). To the prepared PDA media plate, lawn culture of 100 mg/ml) and used as reference standard. A mixture of the fungal pathogen were developed using a sterile cot- 1 ml of methanol and 2 ml of DPPH was used as the con- ton swab of fungal spores. Sterile cork borer were used trol. The reaction was carried out in triplicates and the to make wells and to these wells, different concentration mixture was shaken well, incubated in dark condition for of leaf extracts were added. The plates were incubated at ◦ 30 min after the incubation period he absorbance was mea- 28 C.