INTRODUCTION:

Epilepsy is a group of long-term neurological disorders characterized by epileptic seizures. These seizures are episodes that can vary from brief and nearly undetectable to long periods of vigorous shaking. In epilepsy, seizures tend to recur, and have no immediate underlying cause while seizures that occur due to a specific cause are not deemed to represent epilepsy. In most cases the cause is unknown, although some people develop epilepsy as the result of brain injury, stroke, brain cancer, and drug and alcohol misuse, among others. Epileptic seizures are the result of excessive and abnormal cortical nerve cell activity in the brain. The diagnosis typically involves ruling out other conditions that might cause similar symptoms (such as syncope) as well as figuring out whether any immediate causes are present. Epilepsy can often be confirmed with an electroencephalogram (EEG).

Curcuma amada Roxb. (Family: Zingiberaceae) is perennial rhizomatic aromatic herb which is known as Mango ginger. The mango ginger rhizome was found to be a rich source of fibers and starch. The acetone extract of mango ginger is composed of colourless oil, curcumin, phytosterol and azulenogenic oil containing pinene, camphor, curcumene and ar-turmerone. There are more than 100 phytochemicals reported from fresh and dried extracts of C. amada.

EXPERIMENTAL:

Plant Materials:

The plant has been selected on the basis of its availability and use of the plant. Rhizomes of Curcuma amada were collected from local area of Bhopal in the month of March, 2019. Drying of rhizomes was carried out in sun but under the shade. Dried rhizomes were preserved in plastic bags and closed tightly and powdered as per the requirements. Rhizomes of Curcuma amada were dried and coarsely powdered. They should be extracted with hydro alcoholic solvent.

Preparation of Extract:

Hydro alcoholic extraction:

142gm dried rhizomes of Curcuma amada were subjected to maceration extraction with (400ml) 80% ethanol for 48 hours 87-88. After complete extraction the solvent was evaporated and concentrated to dry residue. % yield was calculated for each extract after drying under vacuum.

Calculation of Percentage Yield:

The percentage yield of yield of each extract was calculated by using formula:

Weight of extract

Percentage yield =––––––––––––––––––––––––– x 100

Weight of powdered drug taken

The extract was subjected to phytochemical investigation to detect different phyto-constituents and pharmacological studies.

Preliminary Phytochemical Investigation of the Extract:

Phytochemical investigation means to investigate the plant material in terms of its active constituents.

Qualitative Chemical Test:

Different methods of identification were used to investigate phyto-constituents present in the rhizomes of Curcuma amada.

In vivo anti- epileptic activity:

Adult male albino Swiss mice (22–25g) were group housed (n=6–10) under a standard 12 h light/dark cycle and controlled conditions of temperature and humidity (25±2°C, 55–65%). Mice received standard rodent chow and water ad libitum. Mice were acclimatized to laboratory conditions for 7 days before carrying out the experiments. All the experiments were carried in a noise-free room between 08.00 to 15.00 h. Separate group (n=6) of mice was used for each set of experiments. The animal studies were approved by the Institutional Animal Ethics Committee (IAEC), constituted for the purpose of control and supervision of experimental animals by Ministry of Environment and Forests, Government of India, New Delhi, India.

Drugs and Chemicals:

Pentylenetetrazole (PTZ) (Hi media Laboratories Pvt. Ltd., Bhopal), Diazepam (Ranbaxy, India) were used in present study. Fresh solution was prepared before each experiment. All other reagents used were standard laboratory reagents of analytical grade and were purchased locally.

Toxicity Study:

Preliminary experiments were carried out on mice (n=6). Hydroalcoholic extract of Curcuma amada were administered orally in different doses to find out the range of doses which cause zero and 100% mortality of animals. Acute oral toxicity was conducted according to the method of Organization for Economic Co-operation and Development (OECD). Animals were kept fasting providing only water, extract were given p. o. in doses of 500, 1000 and 2000mg/kg/ p. o. administered orally for 4 days of different groups of mice (n=6) and the animals were kept under observation for mortality as well as any behavioral changes for evaluation of a possible anti-epileptic effect.

Pentylenetetrazole-Induced Seizures Test:

Mice were divided into three groups each containing six animals, and received hydro-alcoholic extract of Curcuma amada (100 and 200mg/kg) and diazepam (3mg/kg). Thirty minutes later seizures were induced by the pentylenetetrazole (80mg/kg, i.p.). The animals were observed during the first 30 min for number of animals with convulsions i.e. latency and duration of myoclonic jerks, number of deaths and percent protection against convulsion and mortality.

Maximal Electroshock-Induced Seizures Test:

Mice were divided into three groups each containing six animals and treated with either hydro-alcoholic extract of Curcuma amada (100 and 200mg/kg) and diazepam (3 mg/kg). Thirty minutes later seizures were induced by a current stimulus (18 mA, 50 Hz for 0.2 s) delivered by using corneal electrodes by a shock generator (Inco, India). The percent protection and duration of tonic hind limb extension (i.e., the hind limbs of animals outstretched at 180° to the plane of the body axis) was observed. Protection was defined as complete absence of tonic hind limb extension.

RESULTS AND DISCUSSION:

Preliminary phytochemical investigation of the extract:

Percentage Yield of Curcuma amada:

Finally extraction of defatted seeds was done with Hydro alcoholic and % yield was found to be 5.88% w/w and their characteristics are reported in table 1.

Table 1. Physical characteristics of extract

S. No.	Extracts	% Yield
1.	Hydro alcoholic	8.14

Qualitative Chemical Test:

Results obtained from qualitative chemical tests are tabulated in Table 2.

Table-2: Quantitative Chemical Tests of Extract of Curcuma amada

Chemical test

Hydro alcoholic extract

Alkaloids

Wagner’s Test

Hager’s Test

+ve

-ve

Carbohydrates

Fehling’s Test

+ve

Glycosides

Legal’s Test

-ve

Saponins

Froth Test

+ve

Phenol

Ferric Chloride Test

+ve

Flavonoids

Alkaline Reagent Test

Lead acetate Test

+ve

Proteins and aminoacids

Xanthoproteic Test

+ve

Diterpenes

Copper acetate Test

-ve

+ ve – Present, - ve – Absent

Results of in vivo Anti- Epileptic Activity:

Hydroalcoholic extract of Curcuma amada shows antiepileptic property against epilepsy induced by Maximal electroshock (MES), and Pentylenetetrazole (PTZ).

Pentylenetetrazole-Induced Seizure Test:

Table -3: Effects of Hydroalcoholic extract of Curcuma amada on pentylenetetrazole-induced seizures

Treatment	Dose (mg/kg)	Time to seizure onset (s)	% protection against seizures
Diazepam	3	780.0 ± 14.00*	100
Hydroalcoholic extract of Curcuma amada	100	500.02 ± 13.00	64.10
Hydroalcoholic extract of Curcuma amada	200	600.03 ± 15.00*	76.92

Values are expressed as the mean ± SEM of six observations. *P<0.01 vs. saline treatment (One-way ANOVA followed by Dunnett's post hoc test).

Figure 1. Effects of Hydroalcoholic Extract of Curcuma amada on Pentylenetetrazole-Induced Seizures

Maximal Electroshock-Induced (Mes) Seizure Test

Table -4: Effects of Hydroalcoholic extract of Curcuma amada on MES induced seizures

Treatment	Dose (mg/kg)	Time to seizure onset (s)
Diazepam	3	0.00 ± 0.00*
Hydroalcoholic extract of Curcuma amada	100	55.80±2.50
Hydroalcoholic extract of Curcuma amada	200	70.00±2.00*

Values are expressed as the mean ± SEM of six observations. *P<0.01 vs. saline treatment (One-way ANOVA followed by Dunnett's post hoc test).

Figure 2: Effects of Hydroalcoholic extract of Curcuma amada on MES induced seizures

CONCLUSION:

The preliminary phytochemical screening of the hydroalcoholic extract of rhizomes of Curcuma amada revealed the presence of carbohydrates, Proteins and amino acids alkaloids, saponins, phenol and flavonoids. These phytochemical constituents have been reported to be associated with different pharmacological activities of plants. Phenol and flavonoids among other phytochemicals have been reported to possess anticonvulsant activity.

In conclusion, hydroalcoholic extract of Curcuma amada exhibited anticonvulsant potential in various animal models probably because of its neuro-modulatory effect.

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Received on 09.12.2019 Modified on 30.01.2020

Asian J. Research Chem. 2020; 13(1): 19-22.

DOI: 10.5958/0974-4150.2020.00005.X

Millennium College of Pharmacy, Bhopal- 462001

INTRODUCTION: