The Antidiabetic and Spectral Analysis of Various Extracts of Lilium polyphyllum

Background: The Antidiabetic and Spectral Analysis of Various Extracts of Lilium polyphyllum Methods: The study involves antidiabetic and spectral analysis, in which inhibition of alpha amylase and alpha glycosidase inhibition was carried out by various extracts of Lilium polyphyllum. Results: The plant showed a good inhibiting potential of amylase and glycosidase, the two key enzymes responsible for the breakdown of oligosaccharides into glucose molecules. Among the extracts the inhibition of alpha amylase was found to be 54.95% by water extract, 53.01% by ethanol extract and 47.87% by DCM extract. The percentage inhibition of alpha glucosidase was found to be 48.48 % by water extract, 48.48% by ethanol extract and 43.85% by DCM extracts. GC-MS analysis involves isolation of, Methyl 2-furoate, 5-hydroxymethyl furfural, Methyl piperate, Piperine, Palmitic acid, Methyl Palmitate, which in turn justifies the antidiabetic properties of the concerned plant. So the plant can act as good source of antidiabetic drug. The water extract is more beneficial than other extracts. Conclusion: The inhibiting potential of natural products have been found as a good choice for the diabetic patients as they did not led to side effects. Similarly in the above study it had been found that Lilium polyphyllum posses many biosignitures which have the amylase and glycosidase inhibiting potential, among which polar components have more tendency than the less polar.

population of developing countries is directly dependent on traditional medicines, which they consume from plants, for their primary health care needs.
Lilium polyphyllum commonly known as Kshirkakoli or white lilly (family Liliaceae) is extensively used in many indigenous preparations from times immoral. Kshirkakoli have been found to contain Sugar, Alkaloids, flavanoids, and essential Steroids (Subramoniam et.al 2013).
Medicinally, the bulb of this species is used as diuretic, antipyretic and as a energy tonic Few species of the plant are also known for medicinal and food value (Chang C, 2000, Wawrosch C, 2001, which increased its economic importance many folds. In traditional system of medicine, the species reported to restore health immediately and works as antioxidant in the body (Sharma BD, 2005).
Diabetes is considered as one of the fetal causes of death worldwide as per the estimation in every ten second a person dies from diabetes related problems. In 2007, it was reported by (Das AK et al 2008) that diabetes caused 3.5 million deaths globally. Diabetes was considered as 'a disease of rich man' but now it had spread among all masses (Gupta R et al 2007) in India as well in the whole world. Mostly developing countries have large number of diabetic patients but India is considered as infamously dubbed 'diabetic capital of the world' (Abate N et al 2007) because of the largest number of diabetic patients.

Plant materials
The plant material was collected from Kishtwar region of Jammu and Kashmir and was authenticated at FRI Dehradun. The collection process was preferably done in the dry condition.
Plant was shade dried at room temperature and the plant samples were air dried and grounded into uniform powder with a grinder.

Materials and Methods
All the chemicals used in this investigation were of analytical reagent (AR) grade and were purchased from Sigma Merck. De-ionized water was used in the whole study. All the glassware and equipment used for handling were stabilized properly prior to use.

Extraction
The extraction procedure was carried out first with DCM followed by ethanol and then with water based upon their polarity index. The extraction was done by Soxhlet extraction using thimble to get the pure form of extract. 75 g of the plant material was used for extraction.

Inhibition of alpha amylase enzyme
A total of 500 μL of test samples and standard drug (100-1000μg/ml) were added to 500 μl of 0.20 mM phosphate buffer (pH 6.9) containing α-amylase (0.5mg/ml) solution and were incubated at 25°C for 10 min. After these, 500 μL of a 1% starch solution in 0.02 M sodium phosphate buffer (pH 6.9) was added to each tube. The reaction mixtures were then incubated at 25°C for 10 min. The reaction was stopped with 1.0 ml of 3, 5 di-nitro salicylic acid colour reagent. The test tubes were then incubated in a boiling water bath for 5 min, cooled to room temperature. The reaction mixture was then diluted after adding 10 ml distilled water and absorbance was measured at 540 nm. Control represent 100% enzyme activity and were

Inhibition of alpha-glycosidase enzyme
The inhibitory activity was determined by incubating a solution of starch substrate (2 % w/v maltose or sucrose) 1 ml with 0.2 M Tris buffer pH 8.0 and various concentration of plant extract for 5 min at 37°C. The reaction was initiated by adding 1 ml of alpha-glucosidase enzyme (1U/ml) to it followed by incubation for 40 min at 35°C. Then the reaction was terminated by the addition of 2 ml of 6N HCl. Then the intensity of the colour was measured at 540nm (Krishnaveni, S, 1984).

Calculation of 50% Inhibitory Concentration (IC50)
The concentration of the plant extracts required to scavenge 50% of the radicals (IC50) was calculated by using the percentage scavenging activities at five different concentrations of the extract. Percentage inhibition (I %) was calculated by I % = (Ac-As)/Ac X 100 (Shai L, 2010) Where Ac is the absorbance of the control and As is the absorbance of the sample.

Observations and Results
In the study the Acarbose has been used as a standard drug for the inhibition of amylases and glucosidases, and it had been found that the inhibition of the concerned enzymes occur very well in which the maximum inhibition was shown by water extract followed by ethanol and least results were observed for the DCM extracts. The results are shown in the respective tables and a comparative study could be observed from the figures.

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As per the results obtained from the above study it had been found that the inhibition of amylase and glycosidase are dose-dependent, as there is an increase in percentage inhibition in the enzymes by the increase in the concentration of the plant extracts.
The inhibition percentage of alpha amylase was found to be 54.95% by water extract, 53.01% by ethanol extract and 47.87% by DCM extract, similarly the percentage inhibition of alpha glucosidase was found to be 48.48 % by water extract, 48.48% by ethanol extract and 43.85% by DCM extracts. Also as per the comparative study the inhibition of glucosidases enzymes occurs more than the amylases.
The ethanolic extract of the concerned plant when analysed by GC-MS technique the following compounds have been identified.

Discussion
Diabetes mellitus is completely a metabolic disorder and the key factor responsible for it

Conclusion
The study in reference showed that the plant extracts in reference have good inhibiting potential against the amylase and glycosidase enzymes. The inhibiting potential of natural products have been found as a good choice for the diabetic patients as they did not led to side effects. Similarly in the above study it had been found that Lilium polyphyllum posses many biosignitures which have the amylase and glycosidase inhibiting potential, among which polar components have more tendency than the less polar. So the plant in reference with a limited concentration range should be consumed along with the daily routine diet.

Declaration:
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