Malva Parviflora is a leafy vegetable, its family is Malvaceae family exist worldwide including Iraq, where there are naturally in fields and on road edges, the plant is characterized by strong and deep roots and the length of the plant is between 10- 30 cm. The plant is traditionally used to treat all types of infections, especially its use as an anti -hemorrhoid. For the ability to create the second white products such as tannins, alkaloids, that have antibacterial properties [1,2].

Malva parviflora key elements are sterols, flavonoids, polyphenolic compounds and terpenes. In an investigation, phytochemicals including β amyrin, α-amyrin, a mixture of β-sitosterol and stigmasterol, cholesterol, campasterol and ergosterol were separated from the petroleum portion of Malva parviflora. Ethyl acetate fraction yielded kampeferol-3-6’’-p-coumaroyl-O-β-D-glucoside Malva parviflora constituted antioxidants such as ascorbic acid, β-carotenes, αtocopherols and glutathione. It constituted of minerals such as iron, copper, zinc, phosphorous and manganese [3,4].

Inflammation is a response in the organism against types of pathogenic factors by infectious microorganism (bacteria, fungi, toxic chemicals, physical injury or tumors growth) and thus leads to a local accumulation of the plasma liquid [5]. It was also found that the plant has a great effectiveness against bacteria and fungi.as it inhibits the growth of the positive and negative [6].

Research aims to investigate the antimicrobial effectiveness of extracts alcoholic and aquous of malva in different concentrations and comparing with reference antimicrobial agents such as gentamicin and nystatin.

Materials

Leaves were obtained from the local markets and classified by the College of Science/University of Baghdad. Mallow was classified as Malva parviflora, after which it was washed well with water to remove all the dust and impurities stuck in it and left to dry in the shade with constant stirring to prevent it from rotting, the leaves of the plant were ground by the electric grinder to get a fine powder. They were kept in sterile and opaque containers and in refrigerated conditions until use.

Preparation of Plant Extracts

• Aqueous Extract by Soaking: 80 gm of leaf powder was taken and placed in a 1-liter glass beaker, to which 800 mL of distilled water was added. The sterile amalgam was filtered and the solution was filtered with a Buechner funnel with filter paper Whatmman no.1, concentrated employing a rotary vaccum evaporator. The concentrated material was dried in a steel tray in an electric oven at 40°C and then collected. Dry extract and put in an opaque vial until use

• Alcoholic Extract by Soaking: Weigh 80 gm of powder leaves and put it in a 1-liter glass beaker, add 250 mL of ethyl alcohol 80% and place it in the vibrating incubator at a temperature of 37 C for 6 hours, then pass it on several layers of cloth The sterile amalgam was filtered for filtering, then the solution was filtered and concentrated as the previous method. The concentrated material was dried in a steel tray in an electric oven at 40 °C, then the dry extract was collected and placed in an opaque bottle until use

Chemical Detection of Active Compounds 

A set of qualitative tests were conducted to identify the active chemical components of the oil extracts of the cumin plant [7]:

• Tannins Detection: 1mL of aqueous lead acetate 1% was added to 1mL of the extract, a white precipitate as an indication of the presence of tannins 

• Carbohydrate Test: The test was carried out using Molish's reagent, where 1 mL of alcoholic extract was stirred with 5 drops of α-naphthol alcohol, shaken well and then sulfuric acid 2.5 mL was added, when a blue ring was formed indicating the carbohydrates existence

• Glycosides Test: The glycosides were detected by means of Fehling reagent and the forming of a red precipitate indicates the glycosides existence

• Phenols Test: 0.1 gm of the extract was dissolved in 1 mL water, add 1-2 drops of ferric chloride solution FeCl3, blue or green color formation indicates the presence of phenols

• Resins Test: 1 mL of lead acetate 1% was added, a white precipitate formation indicates the presence of resins 

• Detection of Flavonoids: 1 mL of 5 N alcoholic potassium hydroxide was added, a yellow precipitate is formed, indicating the flavonoids existence [8]

• Detection of Saponin Test: 1 mL of aqueous mercury chloride reagent 5% was added to 1 mL of the extract and when a white precipitate is formed, the result is considered positive and indicates the exitence of saponins

• Alkaloid Test: Employing addition several drops of Wagners reagent to 1 mL of the extract, turbidity formation indicating the alkaloids existence 

• Protein Test: Employing 1 mL Biuret reagent, which consists of copper sulfate and potassium sodium tartrate and when the violet color is an indication of the presence of proteins

• Coumarins Test: Coumarins were detected by placing a quantity of the alcoholic extract of the plant in a test tube, then covered it with a filter paper moistened with dilute NaOH solution and heated in a water bath for minutes, then exposed to ultraviolet rays, the color of the paper turns to a bright yellow-green color indicates the coumarin existence

• Detection of Terpenes and Steroids: Mixing 1 g of the extract in a little chloroform and addition of a drop from acetic anhydride and concentrated sulfuric acid. Formation of the brown color, indicating that it contains the terpenes extract, but if it is after a period of dark blue color, it indicates that the extract contains steroids [9]

Determination of the Half-Lethal Dose of the Extracts

The Albino rats were used for this purpose and the suggested circumstances were provided, where the temperature of the animal home was 22 ° C, 30% of humidity, light was supplied for twelve hours a day and the average weight of the animals was twenty-five grams for non-pregnant females. Before administering the doses specified in [10,11], the animals were fasted for three hours. First dose was 50 mg per kilogram of animal weight.

Table 1 revealed the results of the detection of the active substances in the extracts of the leaves of the Iraqi aqueous and alcoholic extract, as this evaluation of the types of detections depends mainly on the components of food metabolism that occur naturally in the cells of medicinal and aromatic plants, which are considered main metabolic products or natural products. These primary compounds or natural products are chemically different in their medicinally active groups, despite the different types of plants that produce these medicinal substances.

Table 1: Chemical Qualitative Tests for Extracts OF Leaves of the Plant Alcoholic and Aqueous

Flavonoids are water-soluble pigments that have most of the therapeutic effects of medicinal plants, as they act as anti-inflammatories, histamines, viruses and antioxidants [12]. Tannins have an effective role in treating burns and wounds and act as antiseptics, as they prevent the growth of microorganisms [13].

The LD50 test, which was conducted on mice, demonstrated that no aberrant symptoms or deaths were observed after administration of the extract made from the plant's leaves at the three concentrations during the first 4 hours. This demonstrates that the aqueous and alcoholic extracts of hibiscus leaves are safe and non-lethal at the doses utilized in the experiment (Table 2).

Table 2: Toxicological Examination of the Aqueous and Alcoholic Extract of the Leaves of the Plant

The results of the elemental examination showed that the water and alcoholic extracts of the Iraqi hibiscus plant contain iron, where the percentage in the aqueous extract was 1.95 and the alcoholic extract was 0.6ppm, as well as the zinc element, where the percentage in the water extract was 1.145ppm and the alcoholic extract was ppm 1.421. The existence of elements, including iron, in the mallow plant confirms that the plant is rich and effective in treating the general weakness of the body and anemia, increasing and supporting the immune system, as well as enhancing energy [14] (Table 3).

Table 3: Concentration of Iron and Zinc Elements in the Leaves of the Plant

Table 3 showed a difference in the rates of the diameters of the inhibition zones according to the sensitivity of each type of bacteria and the type of solvent used and thus the difference in the rate of extraction of the active secondary metabolites present in the plant [15] Where all of Stephylococcus epidermidis, Staphylococcus aureus, Candida albicans, streptococcus mutans were used.

Table 4 showed a difference in the biological activity results of the types of extracts studied, with a difference in the inhibitory role of the tested bacteria compared to Gentamicin and Nystatin control. It showed an inhibitory activity of 19 mm, 15 mm, while the aqueous extract showed a lower activity of the same concentration 16 mm, 14 mm and the activity of the standard antibiotic was 20 mm. Whereas, the aqueous extract of the hibiscus plant at a concentration of 100%, 50% gave an inhibitory activity against Staphylococcus epidermidis 17 mm, 14 mm, while the alcoholic extract of the same concentration showed no activity, while the activity of the standard antibiotic was 17 mm. Also, the alcoholic extract showed an inhibitory activity at a concentration of 100%, 50% on Streptococcus mutants 19 mm, 17 mm, but the aqueous extract showed no activity and the inhibitory activity of the antibiotic was 17 mm. The alcoholic extract of the leaves of the baker plant gave activity against Candida albicans at a concentration of 100%, 50% and the fungus 18 mm, 15 mm, while the aqueous extract gave less activity for the same concentration of inhibition diameter 15 mm, 12 mm and gave the standard antibiotic Nystatin inhibition diameter 16 mm.

Table 4: Biological Activity of Different Kinds of Extracts of the leaves of (Alcoholic and Aqueous) Against Different Kinds of Microorganisms

The presence of alkaloids, phenols, coumarins and tannins has an effective effect in inhibiting inflammatory bacteria and thus helps in the healing process and this confirms the findings of the effectiveness of the aqueous and alcoholic extract on bacteria and Candida albicans species, as well as the superiority of the alcoholic extract over the aqueous extract because it contains a concentration of Higher in groups such as flavonoids and phenols [16].

Depending on the results of the anti-bacterial activity test, the microbial effectiveness of alcoholic extract of the leaves is higher than that of the aqueous extract at 100% concentration and therefore it is considered the best. Furthermore, the results of the toxicological examination of both extracts of the leaves of the plant that it is safe. Benefiting from the high biological activity of the alcoholic extract of the leaves of the mallow plant against bacterial growth, making formulations and ointments and applying them to laboratory animals and then clinically approved to be effective and safe as well.

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