Phytochemical Screening and Toxic Effects of Ambrosia maritima L Ethanolic Extract in Albino Rats

: Ambrosia maritima was traditionally used in folkloric medicine to treatment of rheumatic pains, asthma, and bilharziasis. In this study the plant tested for their secondary metabolites and toxicological effects. Ambrosia maritima showed high presence of alkaloids, steroids and triterpens, moderate presence of flavonoids and tannins, and trace presence of saponins and cumarins. Biochemical and histological examination were done. Rats treated with Ambrosia maritima showed no significant change from the control, except in the ALT, significant decrease was observed. The histological examination of the plant extract showed normal appearance of most sections therefor plants seemed to be safe for oral consumption.


INTRODUCTION
Plants provide a variety of resources that contribute to the fundamental needs of food, clothing and shelter. Among plants of economic importance are medicinal plants. Medicinal plants are groups of plants with vital roles in alleviating human suffering (Baquar, 2001).
Plants have been utilized as therapeutic agents since time immemorial in both organized and unorganized forms (Girach et al. 2003).
Information about medicinal plants has been transmitted gradually from generation to generation; a human knowledge has gradually become complete with the formation of civilizations and the provision of more facilities. Medicinal plants used as a medical resource in almost all cultures. Ensuring the safety, quality and effectiveness of medicinal plants and herbal drugs very recently became a key issue in industrialized and developing countries. By standardizing and evaluating the health of active plant-derived compounds, herbal drugs can help the emergence of a new era of the healthcare system to treat human diseases in the future. Awareness of traditional knowledge and medicinal plants can play a key role in the exploitation and discovery of natural plant resources (Jamshidi et al. 2018).The traditional system of treatment, differing in concept and protocol, exemplifies well-developed systems such as allopathic, homeopathic, Ayurvedic, and Chinese systems of treatment. Most of the civilized nations have developed their own Materia Medica, compiling details about various plants used for therapeutic purposes (Khan, 2014).
Safety is the major criterion for indication herbal medicines to use in therapeutic purposes. Toxic potentials closely associated with screening of plants extracts for their activities against microorganism (Bulus et al. 2011). Potential Chemicals that are used in commerce, the home, the environment, and medical practice may present various types of harmful effects. The nature of these effects is determined by the physicochemical characteristics of the agent, its ability to interact with biological systems (hazard), and its potential to come into contact with biological systems (exposure). (Dekant, and Vamvakas, 2000). Some medicinal plants have unpleasant side effects which may be attributed to over doses or other factors. This may lead to acute toxicity and death, however, when these problems are well treated, the therapeutic potentials of medicinal and aromatic plants will harness for further drug development in the future. (Okigbo et al. 2009). Modern medicines have little to offer for alleviation of hepatic diseases and it is chiefly the plant based preparations which are employed for their treatment of liver disorders. However, there is not much drugs available for the treatment of liver disorders Therefore, many folk remedies from plant origin are tested for its potential activities in experimental animal model (Chacko et al. 2012).
Ambrosia maritim family Asteraceae. annual or short-lived perennial plant (Hammouda, et al. 2009). Is known locally as "Damsissa" (Fig. 1). It is a widely distributed weed especially near water catchment. The species was found in Sudan in Nile bank, on muddy canal banks, and Egypt, it grows in Nile delta, south Sinia (Makkawi et al. 2015).And Oases and Mediterranean region (Hammouda, et al. 2009). It is a common folk medicine used in the treatment of rheumatic pains, asthma, and bilharziasis. (Helal et al. 2014) Plants extracts should not only be efficacious but safe for consumption, therefore this study aimed to evaluate the toxic effects of the plant under investigation in order to confirm the safe for oral dose.

Plant Materials and extraction
Ambrosia maritima (Damsissa) was obtained from Khartoum state, the plant was identified at the Herbarium, Institute of medical and aromatic plants. Extraction was carried out according to method described by (Sukhdev et al. 2008). The plant sample was coarsely powdered using mortar and pestle. Then extracted by soaking in 80 % ethanol for about five days with daily filtration and evaporation. Solvent was evaporated to dryness by using rotary evaporator and the extract of each part combined together.

Phytochemical screening
Phytochemical screening for the active constituents was carried out for ethanolic extract using the methods described by (Martinez et al., 2003), Sofowora (1993), Harborne (1984 and Wall et al. (1952) with many few modifications.

Experimental design
Twenty four white male albino rats were obtained from the Institute of medical and aromatic plants _ Sudan; the animals were divided to (2) groups. Group 1 the control contain 6 rats which orally dosed with distilled water, group 2 contain 18 rats divided into 3 sub-groups which orally dosed with extract of plant (450, 800, and 1400mg/kg) for 14 days.

Blood Sampling and Processing
At the end of the experiment (14 days) the rats were decapitated and blood samples were obtained from each rat, the samples were collected in plain tube to obtain serum; blood was left for one hour to clot and the tube was centrifuged at 3000 rpm for 15 minutes and the harvested serum was used for biochemical analysis. The rats were quickly dissected and the whole liver was excised, preserved in a formalin solution. Tissues sectioning were made, stained and histopathological examination was done.

Biochemical Parameters
Serum samples were analyzed for the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and for the concentrations of total protein, albumin, total bilirubin, alkaline phosphate and urea by the Roche Diagnostic Hitachi 902 Analyzer. Assay kits (Bio Systems S. A.Costa Brava, 30, 08030 Barcelona Spain) were employed for the analyses, the parameters were determined in the serum following the procedure described in the kits.

STATISTICAL ANALYSIS
Analysis of variance (ANOVA) for completely randomized design with six replicates was used followed by Duncan multiple range test (DMRT) to detect significant differences in all the parameters, Values were considered statistically significant at p < 0.05 (Gomez and Gomez 1984).

Phytochemical screening
The plant under investigation (_Ambrosia maritima, Damsissa) tested for secondary metabolites as described by Harborne (1984). The results indicated high presence of alkaloids, steroids and triterpens, moderate presence of flavonoids and tannins, trace presence of saponins and Cumarins, the result agreement with what was found by EL-Kamali et al. (2010).Also Helal et al. (2014) reported presence of all secondary metabolites.

Biochemical Finding
The present study highlight the effect of toxicity of (Ambrosia maritima) extract on liver function (LF) parameters (ALT, AST, ALP, protein, albumin, globulin and bilirubin) of albino rats which were investigated. The administrated levels were 450 mg /kg (low dose), 800mg/kg (medium dose) and 1400mg/kg (high dose), in addition to the control. Six randomly albino rats were assigned for each group (treatment). Table2 showed that the level of Alanine Amino Transferase (ALT) in rats was not significantly different between the studied extract doses of (Ambrosia maritima) species (450, 800, and 1400mg/kg ) and control as well as between the doses themselves and it was ranged between 18.97 U/L for control and 17.89 U/L for the high dose (1400mg/kg). Also (AST) was not significantly affected by admitted levels of Ambrosia maritima extract the values were (71.97, 64.87 and 72.60 U/L for low, medium and high doses, respectively) than in control (85.82 U/L) by about 16.1%, 24.4% and 15.4%, respectively.  EL muoiz et al., (2020) presented significant reduction on AST level and no change on ALT and ALP. Also they reported that A. maritima has many compounds that lead to improvement the hepatocytes and preserve the structural of hepaticcellular membranes.
Moreover the values of protein, albumin, globulin and bilirubin were not significantly different from the control. The results for ALT,AST and ALP in this study were agreement with Ahmed et al., ( 2001) who indicated hepatoprotective result when they used methanolic extract of A. maritima against acetaminophen (paracetamol,4hydroxy acetalline). Also A.maritima has not got toxic signs could be detected after oral administration of methanolic extract, during four weeks (Alard, 1991).