Research Article Open Acc J of Toxicol Volume 4 Issue 1 - April 2019 Copyright © All rights are reserved by Salem Mohamed Edrah DOI : 10.19080/OAJT.2019.04.555626 Phytochemical Study and In Vitro Antibacterial Activity of Two Traditional Medicinal Plants (Vinca Rosea and Vinca Difformis) from Libya Salem Mohamed Edrah1*, Fatimh Mustafa Meelad1 and Fouzy Alafid2 1Department of Chemistry, Al-Khums El-Mergeb University, Libya 2University of Pardubice, Czech Republic Submission: March 04, 2019; Published: April 02, 2019 *Corresponding author: Salem Mohamed Edrah, Department of Chemistry, Faculty of Sciences Al-Khums El-Mergeb University, Al-Khums, Libya Abstract Vinca rosea and Vinca difformis gardens. Aqueous and Ethanol extracts leaf of both species were preliminary phytochemically screened and tested against some pathogenic bacteria. Qualitatively analysis revealed are magnificentTannin, Saponin, plants Flavonoids growing in and the Terpenoids Libyan woodlands gave positive and are results utilized and beautifully phlobatanins as an and embellishing Steroids gave in negative results. Quantitative analysis revealed that the percentage yield of bioactive constituents is present more in Vinca rosea than in Vinca difformis. In addition, the crude extracts of Vinca rosea and Vinca difformis were tested (using the Disc Diffusion Method) for their antimicrobial bacteria include Staphylococcus aureus and Streptococcus spp., and two strains of gram-negative bacteria including Escherichia coli and Klebsiella pneumoniaactivity against the bacterial pathogens. The influences of aqueous and ethanol extracts on some pathogenic: two strains of gram-positive antibiotics by measuring the diameter of the inhibition zones, After incubation, the zone of inhibition was measured in mm, a good inhibition of more than ,6 and mm the were sensitivity observed of indicating the microorganisms the effective to antibacterial the extracts activityof plant’s of thespecies bioactive were compoundscompared with in both each of other the plants and with leaves designated extracts. Moreover, the including results of the antibacterial activities of plants leaves extracts were discussed regarding their phytochemical components which exhibited that the Vinca rosea and Vinca difformis difformis have a good inhibition zone against two tested bacterial strains. Keywords: Vinca rosea; Vinca difformis; Bioactive Constituents; Qualitative and Quantitative analysis; Antibacterial activities Introduction a scenically and medicinal plant, as a source of the drugs which Medical plants are usually used in conventional medicine as used to treat cancer. Because of presence bioactive ingredients treatments for several diseases and contagious illnesses. Utilize which are existing in plants such as phenolic, tannins, alkaloids herbal plants own fewer side consequences and including its cost - - dants owing to existing these substantive useful composites, and portant role in the process of public and private parks, which are and flavonoids are vital and characterized as a natural’s antioxi is lower. Herbal plants are a group of flower plants that play an im which effect on improvement of the performance of humane body multi-color, shapes, sizes plant. Vinca is one of the grass groups of high coordination value for its numerous forms. It has multi- certain excretion of hormones. Moreover, Vinca rosea assistance ple uses in the garden and can be used in designing and streets organs, adding, are entered in modifiable of lowering or raising in cumulative the insulin creation which helps in healing diabetes, to withstand extreme external conditions. Vinca rosea and Vinca - difformis abetic potential of this plant through using crude extracts rather utilized beautifully as an embellishing in gardens, they have a furthermore, the major influence complete is around the antidi are magnificent plants growing in the woodlands and more than the pure bioactive compounds [1,2]. blooming season from late winter to early spring and are often ev- ergreen. Vinca rosea Vinca difform- Materials and Methods is Catharanthus roseus, ordinarily has purplish red flowers while Collection and extraction known as Vinca rosea and recognized as the rose periwinkle, rosy has whitish-blue flowers. The periwinkle or Madagascar periwinkle. Though the Vinca difform The fresh and healthy samples of leaves of Vinca rosea and is commonly called the intermediate periwinkle, is a species of Vinca difformis were collected from wild of Al-Khums, Al-Khums, Apocynaceae. Catharanthus Libya. They were washed by tap water then by distilled water, lat- roseus original and endemic to Madagascar were bout species are flowering plant in the dogbane family growing in Europe, North Africa and south-west Asia, and used as °C er that dried in shade, drying then was finished in an oven at 45 . Samples were powered by the electrical blender. 20g of finely Open Acc J of Toxicol. 4(1): OAJT.MS.ID.555626 (2019) 001 Open Access Journal of Toxicology powdered leaves of the Vinca rosea and Vinca difformis whereas of the test tube. The appearance of a violet ring at the interphase each sample was soaked with 400ml appropriate solvent (dis- indicated the presence of carbohydrate. Iodine test: 5ml of the crude extract was mixed with 2ml of on the shaker and then settled at a room temperature 25±2 °C for tilled water and ethanol, separately), into flasks which were fixed iodine solution. A dark blue or purple colouration indicated the presence of the carbohydrate. pressure with the help of rotary vacuum evaporator to yield a vis- 72 hours. The solvent was filtered then removed at the reduced cous dark brown residue of water extract and a viscous dark green Flavonoids residue of ethanol extract. Shinoda test: 5ml of the crude extract was mixed with few Qualitative phytochemical screening fragments of magnesium powder. Concentrated HCl was added dropwise. Pink scarlet color appeared after a few minutes which Vinca rosea and Vinca dif- formis and the prepared crude extracts were carried out for the The finely powdered leaves of the existence of bioactive composites by via standard methods [3-7]. indicatedAlkaline the presencereagent testof flavonoids.: 5ml of the crude extract was mixed with 2ml of 2% solution of NaOH. An intense yellow color was Steroid formed which turned colorless on the addition of a few drops of 5ml of crude extract was mixed with 2ml of chloroform and concentrated sulphuric acid was added carefully along the side of dilutedSaponins HCl acid which indicated the presence of flavonoids. the test tube. A red color formed in the lower chloroform layer indicated the presence of steroids. 5ml of distilled water was mixed with extract in a test tube and was shaken vigorously. The formation of stable foam was tak- Sterols and triterpenes en as an indication for the presence of saponins. Alkaloids 20ml of 50% alcohol was added, the tube was then heated for 5g of the finely powdered leaves was sited in a test tube and 3min in a water bath. It was then allowed to cool to room tem- - with 50ml of 5% H2S04 - 5g of the finely powdered leaves were boiled in a water bath orating beaker to dryness and about 10ml of petroleum ether was perature and filtered. The filtrate was then evaporated in an evap in 50% ethanol. Then was cooled and fil added to the beaker and stirred for 5min, the petroleum ether put in 200ml of separating funnel and the solution was made al- tered. A portion was set aside. One more portion of the filtrate was portion was then discarded. 15ml of chloroform was then add- kaline by adding 3 drops of concentrated ammonia solution. For- ed and stirred for about 5min, it was then transferred into a test merly Equal volume of chloroform was added and shaken gently tube and about 1mg of anhydrous sodium sulphate was added and to allow the layer to separate. The lower chloroform layer was col- lected into a second separating funnel. The ammoniacal layer was test tubes and used for the following tests set aside. The chloroform layer was extracted with two quantities shaken gently and filtered, the filtrate was then divided into two each of about 15ml of dilute sulphuric acid. The various extracts a. Salwoski’s test: were then used for the following test: concentrated sulphuric acid was added to form a lower layer. Red- To the first test tube: 2 to 3 drops of dish-brown color at the interphase indicates the existence of a a. Wagner’s test: 2ml of the - steroidal ring. - filtrate was added 1ml of Wag tate indicates the presence of alkaloids. b. Lieberman-Burchard’s reaction: To the second test ner’s reagent in dropwise. Formation of a reddish-brown precipi tube: an equal volume of acetic anhydride was added and gently b. Dragendorff’s test: 2ml of the mixed. Then 1ml of concentrated H S0 was added down the side - 2 4 filtrate was added a of the tube. The appearance of a brownish-red ring at the contact dish-brown precipitate indicates the presence of alkaloids. 1ml of Dragendorff’s reagent in dropwise. Formation of a red zone of the two liquids and a greenish color in the separation layer c. Mayer’s test: 2ml of the indicates the existence of sterols and triterpenes. filtrate was added a 1ml of Carbohydrates cream precipitate indicates the presence of alkaloids. Mayer’s reagent in dropwise. Formation of a greenish colored or Fehling’s test: 2ml of Fehling A and B reagents (Equal volume Tannins and phenols were mixed together) was added to 5ml of crude extract and 5ml of the crude extract was mixed with 1ml of 2% solution of gently boiled. A brick-red precipitate appeared at the bottom of FeCl . A blue-green or black colouration indicated the presence of the test tube indicated the presence of reducing sugars. 3 phenols and tannins. Benedict’s: of crude extract and boiled, a reddish brown precipitate formed Proteins and amino acids 2ml of Benedict’s reagent was mixed with 5ml which indicated the presence of the carbohydrates.