1. INTRODUCTION:

The means word snake has intrigued mankind for years and excite each response. The simple attention to species is due to the physiological effect of their poison on prey such as paralysis, and often death. Envenomation snake bite is a general problem in tropical and subtropical regions' population which are most affected. The South-East Asia is most affected region in the world in terms of population density and agricultural practices according to world health organization (WHO) [1]. From all regions of the world, snake bite injuries are still life threatening and require intensive care, such as malaria, dengue hemorrhagic fever, tuberculosis and parasitic diseases, snakes bite is always a threat [2, 3]. The number of bites of these poisonous snakes is about 1000 with 10 deaths per year non-stop [4]. The formation of snake venom varies according to their habitats within species. It differs on the basis of diet, age, and season. This variation may be responsible for the complexity of the poison.

The most toxic and complex of all natural toxins is poison venom of any nature possess many kinds [3]. The important poisons in humans that affect the nervous system, cardiovascular systems, thrombosis, and tissue necrosis [6]. The doctors' face difficult to diagnose and treat toxic snake bites is the insufficient information, within the management of anti-toxin therapy, is not provided in clinical practice.

Many people currently rely on plants extract and their primary source of medicines in rural areas of the world [7-9]. WHO supported the methods use of local medicines and launched a strategy in 2002 as well as in the period 2014-2023, for proper development and use in the world, including pure extracts and compounds. Few ethnics have led to improve drugs, for further development throughout the world [10-12]. The use of plant preparation as sources of drugs is based on experience and superstition that passed from generation to present [13-16]. Research on medicinal plants in recent times was increased throughout the world to be use in different diseases [17-19], because they have biological effects against many diseases and have been using in traditional medicine [20-23]. But, many medicinal plants have not been reported scientifically on their biological importance. There were no earlier reports on anticoagulation of the current selected medicinal plants. So, the current study was aimed to compare the commonly available medicinal plants in tropical region to against snake venom.

2. MATERIALS AND METHODS:

Plant materials collection and preparation of extracts:

The plant materials Allium sativum, Allium cepa and Nigella sativa were collected from the local market. The collected material were washed cleanly and dried under shade. The dried material was used to prepare aqueous extracts with double distilled water. The dried extracts were stored in desiccator for further use.

Phytochemical screening:

The selected plant extracts were screened to know their phytochemical constituents in them using standard methods [24].

In-vitro DPPH free radical scavenging activity:

The scavenging activity for DPPH free radicals was measured according to the procedure described by Braca et al. In DPPH assay method is based on the reduction of alcoholic DPPH solution (dark blue in color) in the presence of a hydrogen donating antioxidant converted to the non radical form of yellow colored diphenyl-picrylhydrazine. Lower the absorbance higher the free radical scavenging activity [24-26].

In-vitro anti-coagulation activity:

The extracts of Allium sativum, Allium cepa and Nigella sativa at different concentrations i.e. 20, 40 and 60 mg/mL, on a glass prepared 24 hours ago. The weight of the Heparin 1 gm powder was solved in regular saline 5ml and placed separately. After that, 5 ml of blood was pulled by the syringe from a healthy donor's arm and then put half ml of blood in all the prepared concentrations. A drop of poison snake was prepared in advance for all materials prepared and then recorded clotting time for all plants extracts and heparin to measure the time coagulation of the biological activity record of all the prepared substances that lead to slow clotting process and the efficacy of delayed blood clotting [21].

3. RESULTS AND DISCUSSION:

The extracts of selected plants were screened for to know their phytochemical constituent. The extracts showed different chemical constituents in them during analysis. All three plants have different chemical components in them. The three plant extracts showed the presence of phenols, steroids, terpenoids, glycosides, carbohydrates, alkaloids. The extracts gave negative results to amino acids, oils, quinones and saponins. The Allium cepa and Nigella sativa gave positive results to flavanoids and tannins but negative in Allium sativum (Table 1).

The selected plant extracts showed the concentration dependent DPPH free radical scavenging activity. Among three plants, Allium sativum extract have showed more reduction on DPPH radicals' production at 600µg concentration. The percentage inhibition of selected plant extracts was comparable with the standard drug ascorbic acid (Table 2).

Table: 1 Phytochemical constituents present in Allium sativum, Allium cepa and Nigella sativa

Name of the Phytochemicals	Name of the plant
Name of the Phytochemicals	*Allium sativum*	*Allium cepa*	*Nigella sativa*
Phenols	+	+	+
Phytosterols	+	+	+
Terpenoids	+	+	+
Glycosides	+	+	+
Saponins	-	-	-
Flavonoids	-	+	+
Tannins	-	+	+
Carbohydrates	+	+	+
Alkaloids	+	+	+
Amino acids	-	-	-
Oils	-	-	-
Quinones	-	-	-

+ = Present, – = Absent

The selected plant extracts were tested for their anti-coagulation time on blood when treated with snake poision by comapring with the standard anti-coagulant heparin. The different concentration of extract has affected the time delay on coagulation of blood. As the concentration was increased the coagulation time was increased. (Table 3, 4 and 5). The Allium cepa extract showed more delay time on coagulation of the blood (Fig 1).

Table: 2 Concentration dependent percent inhibition of DPPH radical by Allium sativum, Allium cepa and Nigella sativa in In-vitro studies

Name of the plant	Percentage inhibition of DPPH radical
	Quantity of extracts/ ascorbic acid in micrograms (µg)
	50	100	200	400	600
Allium sativum	30.05±1.02	44.22±0.6	52.36±0.82	62.35±0.12	75.8±0.18
Allium cepa	27.25±0.08	34.58±0.48	43.51±0.23	51.39±0.42	65.48±0.22
Nigella sativa	26.40±0.45	36.55±0.32	44.78±0.16	52.68±1.02	64.18±0.44
Ascorbic acid	50.08±0.5	60.35±0.28	72.35±0.01	81.05±0.11	90.44±1.08

Table: 3 Coagulation time of plants extracts in 20% concentration

Name of the plant extract	Coagulation time (minute)
Allium sativum	19.2
Allium cepa	20.7
Nigella sativa	19.6
Heparin	66

Table: 4 Coagulation time of plants extracts in 40% concentration

Type of extracts	Coagulation time (minute)
Allium sativum	20.5
Allium cepa	21.4
Nigella sativa	20.4
Heparin	66

Table: 5 Coagulation time of plants extracts in 60% concentration

Type of extracts	Coagulation time (minute)
Allium sativum	25.7
Allium cepa	29.1
Nigella sativa	26.2
Heparin	66

Fig. 1: Effectiveness of plant extracts on coagulation time

CONCLUSION:

A snake bite has led to high rates of death, especially in rural areas [27]. There is a need to implement a policy to provide rural areas of the world. Given the diversity of species, it is difficult to develop a certain type of immune treatment that has led to the search for alternative sources. Herbal antidote provides the arena to explore as it is abundant and readily available in remote places also [22, 23]. The need enforced use to investigate easily available plant products in all areas around the world to know about their anti-coagulation nature based on knowledge of ancient culture and traditional practices. Numerous studies have been carried out in this regard, and a few new compounds have been isolated. There are still many medicinal plants and many resources still available and need to be studied to isolate the antivenin that can replace the Anti-Snake venoms.

ACKNOWLEDGMENTS:

The authors are thankful to Chemistry Department, College of Science, University of Baghdad for providing the laboratory facilities.

CONFLICT OF INTEREST:

None to declare.

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Received on 08.02.2020 Modified on 04.08.2020

Research J. Pharm. and Tech. 2021; 14(6):2915-2918.

DOI: 10.52711/0974-360X.2021.00511