Cytotoxicity Assessment of Malva Sylvestris Crude Extract on Melanoma and Lymphoma Cell Lines Raghda Rayssan & Muayad S. Shawkat Department of biotechnology, college of science, university of Baghdad, Iraq. Abstract This article focus on the analysis of M.sylvestris methanolic leaves extract by GC-MS and the evaluation of its cytotoxic effect on two types of human cancerous cell lines which are melanoma and lymphoma cell lines. GC-MS analysis showed 29 peaks, the prominent peak was 22.942 peak area (RT 27.01): trans-Phytol; Cyclohexanol,5-methyl-2-(1-methylethyl)-,[1S-(1.alpha.2.beta.5.beta.)]; Dihydrogeraniol; Menthol,trans- 1,3,trans-1,4; (+)-Isomenthol . The extract showed cytotoxicity against both melanoma and lymphoma cell lines and the cytotoxic effect was increased with the increasing of extract concentration, the cytotoxicity of the extract at 200 μL was 68.65% and 76.53% for lymphoma and melanoma cell lines respectively, for reference cell line the cytotoxicity of the extract was 7% at 200 μL. This implicit that M.sylvestris extract has minimum side effect on normal cells thus may be considered safe and potential candidate as anticancer agent. Keywords-Lymphoma, melanoma, cytotoxicity, Malva sylvestris, maceration. INTRODUCTION Cancer is one of the lethal diseases it is characterized via the irregular cell proliferation. The very common reason for cancer is lifestyle changes, therefore there is urgent requirement to discover a better treatment for this lethal disease (According to World Health Organization) [1]. High fatality and incidence implicit it as a significant public health and economical issue that requires an effective way for prevention. Medicinal plants owns many advantages than chemical products because the compounds that are derived from plants are more tolerant with no toxic effect to the normal cells of the human body [2]. Several pharmacological roles of these plants compounds includes antioxidant, antimicrobial, antiviral, anticancer, antifungal and anti-parasitic roles [3]. The available classic curatives for cancer therapy are radiotherapy and chemotherapy both have diverse side effects such as neurological effect, cardiac effect, renal and pulmonary toxicity effect sorely affecting the healthiness of the person. Thus a substitutional method for treatment is in demand that include development of anticancer drug that is less toxic and more potent as compare to the already available drugs in the market. Many studies have been done on naturally occurring plant compounds known to own cytotoxicity effects as they demonstrate potentiality to destroy cancerous cells. On account of the advantages of medicinal plants they are on top of demand and various species have been scanned and selected for the accommodation of cancer medicines. Newly, there has been an elevated interest in the study of compounds from a plant source as anticancer compounds by the concerned scientists. Many studies have showed the role of these plants in prevention and therapy of cancer [4]. presently, near 25% of the drugs produced in all around the world are extracted from plants in a direct way or at least one of its active ingredient is from plant origin, According to world health organization 80% of the world's population depend on plants derived drugs for treatment [5], Also WHO promotes the herbal derived drugs addition in health care programs because these drugs are easy to access at a low cost within the reach of the common people and are time tested, thus counted to be much safer comparing to up to date synthetic drugs [6]. Malva sylvestris is one of the medicinal herbs that used in both food and medicine, this plant characterized by a perennial root, a juicy annual stem of 2-3 height, large heart-shaped seven-lobed leaves, and the Flowers is closely similar to that of honeysuckle [7]. Nowadays, the consumption of this plant is broad spread because modern researches have revealed the important medicinal properties of this plant such as; anti-ulcerogenic, antioxidant, anticancer, and anti-inflammatory [8; 9; 10; 11; 12; 13; 14; 15; 16; 17]. This research is aimed to investigate the cytotoxic activity of the crude methanolic extract of M.sylvestris leaves against two cancerous human cell lines which are melanoma and lymphoma. MATERIALS AND METHODS Collecting plant material and extract preparation The plant material (leaves) of M.sylvestris were collected from Baghdad university gardens and identified by the herbarium in the department of biology/college of science/ university of Baghdad. The plant leaves was cleaned well, washed with tap water and then in distilled water, then wiped and dried in a ventilated cool place away from the sun. After drying the plant material were crushed using a grinder and kept until further usage. The raw plant extract is obtained by maceration which is leaving the powder of the plant leaves in prolonged contact with a solvent (methanol). The separation done by filtration, the powder was macerated for 24 hours at ambient temperature in a mixture of methanol and water (80/20, V/V), the extraction repeated three times with renewal of the solvent, then the macerates were combined then filtered by a filter paper and the solvent was removed from the filtrate under vacuum at 45 o C by a Rota vapor [18]. Chemical detection of secondary metabolites Test for tannins (Braymer’s test): 1ml of distilled water was added for diluting the extract sample and then two drops of ferric chloride was added to the sample. A transient greenish to black color indicates the presence of tannins [19]. Test for saponins (foam test): Small quantity of the extract was diluted with 4ml of distilled water, then the mixture was shaken. Persistence of foam for 10 minutes indicates the presence of saponins [20]. Test for terpenoids (Salkwoski test): The extract was mixed with 0.4ml of chloroform, then 0.6ml of concentrated H 2 SO 4 was carefully added to the sample to form a layer. A reddish brown color at the interface is formed as a positive result for the presence of terpenoids [21]. Test for flavonoids (NAOH Test): Few drops of sodium hydroxide solution added to the extract sample. Formation of intense yellow color, which turn colorless after the addition of acid indicates the presence of flavonoids [22]. Test for phenol (ferric chloride Test): Addition of few drops of ferric chloride solution to the extract sample. Formation of bluish to black color indicates the presence of phenols [23]. Test for Alkaloids (Mayer’s Test): 2% H 2 SO 4 was added to 1ml of the extract sample and warmed for two minutes, then filtered and few drops of mayer’s reagent were added. A creamy-white color precipitation is a positive test [24]. Raghda Rayssan et al /J. Pharm. Sci. & Res. Vol. 11(1), 2019, 70-74 70