INTRODUCTION:

Type 2 diabetes mellitus (DM) is a chronic metabolic disorder in which prevalence has been increasing steadily all over the world. As a result of this trend, it is fast becoming an epidemic in some countries of the world with the number of people affected expected to double in the next decade due to increase in ageing population, thereby adding to the already existing burden for healthcare providers, especially in poorly developed countries.¹

In modern days, allopathic medicines can cause unwanted side effects and sometimes it may react with other medicines and result in adverse effects to a patient. Allopathic drugs do not cure; they just suppress the disease effect for a short period. While herbal drugs have minimum side effects because all the ingredients are naturally made that do not cause devastating effects on the health. So, in this study we evaluated that anti diabetic effect in Syzygium cuminii (Jamun), Gymnema sylvestre (madhunashini), and Swertia chirata (chirata) was found to be effective and beneficial in treatment of diabetes.

It has been studied that Syzynium cuminii (Jamun) seeds of family Myrtaceaeare claimed to contain alkaloid, jambosine, and glycoside jambolin or antimellin, which halts the diastatic conversion of starch into sugar and seed extract has lowered blood sugar.

In Gymnema sylvestre (madhunashini) belonging to the family Asclepiadaceae, presence of gymnemic acid lowers and balance blood sugar level. It blocks the sugar absorption in small intestine and stimulates the beta-cells of pancreas.^2,3

Swertia chirata (chirata) of family Gentianaceae, mangiferin is a xanthonoid show the anti-diabetic properties by increasing glucose uptake and reducing insulin resistance in animal studies. Oleanolic acid is a triterpenoid that shown to have anti diabetic action by glucose uptake.^4,5

All these drugs have been already proved for its anti-diabetic activity. In this study we made an attempt to combine Syzygium cuminii, Gymnema sylvestre, and Swertia chirata to obtain anti diabetic activity which was evaluated using alpha-amylase studies.

MATERIALS AND METHOD:

Preparation of plant extracts: Fresh seeds of Syzygium cuminii, fresh leaves of Gymnema sylvestre and fresh leaves of Swertia chirata were shade dried and pulverized into a fine powder with the help of pestle and mortar. Their powdered forms were put in respective amount of methanol and kept at room temperature overnight and then filtered using Whatman filter paper. The filtrates were then sonicated and the gummy extracts were stored.

Evaluation of granules:

Angle of repose: the powder is allowed to pass from a funnel on a level surface. Measure the height of the peak of powder from ground. Draw a circle around the surface where the powder spreads and calculate the angle of repose.⁶

Table No.1: Angle of repose

Description	Repose Angle
Very free-flowing	<30°
Free flowing	30–38°
Fair to passable flow	38–45°
Cohesive	45–55°
Very cohesive (non-flowing)	>55°

Bulk density: bulk density is determined by pouring presieved bulk drug into a graduated cylinder via a large funnel and measuring the volume and weight.

Weight of powder

Bulk Density = -----------------------------

Volume of powder

Tap density: Tapped density is determined by placing a graduated cylinder containing known mass of drug or formulation on a mechanical tapper apparatus, which is operated for a fixed number of taps (approx. 1000) until the powder bed volume reached a minimum. Using the weight of drug in the cylinder and this minimum volume, the tapped density may be computed.

Weight of powder

Tap density = ----------------------------

Tapped volume

Hausner’s ratio: The Hausner’s ratio is defined as the ratio between the tapped bulk density and the aerated bulk density.

Tap density

Hausner’s ratio = ---------------------

Bulk Density

Carr’s index was calculated using following formula:

Tapped density – Bulk density

Carr’s index (%)= -------------------------------------- × 100

Tapped density

Table No. 2: Composition of tablet

Ingredients	Quantity (in mg)
Jamun	100
Chirayat	200
Madhunashini	150
Starch	40
TALC	10

Preparation of starch paste:

5grams of potato insoluble starch was weighed. The weighed amount of starch was then added in 100ml water in a beaker. Further the mixture was divided into 2 equal halves. One half was heated until its 50% volume. To the above 50% capacity the other divided volume was added. 5% starch paste for binding of the powder was prepared.

Preparation of tablet:

To the weighed and fully powdered mixture of drugs and excipients starch paste solution was added until dough mass was formed. The dough was passed from sieve Number 10 so that granules of the powder were obtained The granules were then sundried for 2-3hours so that complete dry granules were obtained. Then granules were passed from sieve number 10. Dried granules were obtained. Tablets were punched using 10mm punch.

Alpha amylase inhibition assay procedure:

Preparation of reagents:

· Buffer solution: Prepare a solution containing 2.4mg/ml of Sodium phosphate monobasic and 0.39 mg/ml of NaCl, with water adjust to pH 6-9 at 20 degree centigrade using 1N NaOH/1MHCl.

· Starch solution: 1% W/V

· 5.3M Potassium sodium tartrate tetrahydrate solution: Prepare 1,496mg/ml solution of potassium sodium tartrate tetrahydrate in 2M NaOH solution. Dissolve solids by heating on a heating/stir plate with mixing do not heat to boil.

· 96mm Dinitro salicylic acid solution: 21.9mg/ml solution in water.

· Colour reagent solution (for 40ml): 12ml of warm water. (50-70 degree centigrade) + 8ml warm 5.3M potassium sodium tartrate tetra hydrate solution + 20ml warm 96mm dinitrosalicylic acid solution.

· 0.2% (w/v) maltose standard: (2mg/ml)

· Alpha – amylase solution: Immediately before use, prepare a solution containing 0.75-1.5unit/ml of alpha – amylase in 20-degree centigrade water.

Assay procedure:

In 50ml volumetric flask 0.3ml extract, 0.5ml starch solution, 0.5ml amylase solutions were added. The flasks were kept in dark for 10 minutes. 1ml DNS-2 solution was added and boiled for 5 minutes. The resultant mixture was cooled and 3ml distilled water was added. The absorbance was calculated at 540nm using colorimeter.⁷

RESULT AND DISCUSSIONS:

In the present study, three indigenous anti diabetic medicinal plants from India were screened for their alpha-amylase inhibitory potential.

Table No. 3: List of plants used in the study along with the parts used.

Plant Species	Part of plant used	Extracts (methanol extract)
Syzygium cuminii	Seeds	+
Gymnema sylvestre	Leaves	+
Swertia chirata	Leaves	+

+: Inhibitory action against alpha-amylase

-: No inhibitory action against alpha amylase

Evaluation of granules:

The evaluation of granules was done by calculating the following

Table No.4: Result of evaluation of granule

Parameters	Result
Angle of repose	44
Bulk density	1.79
Tap density	2.33
Hausner’s ratio	1.301
Carr’s index	23.17

QC test for tablet:

Weight variation was found to be 0.2391%

Friability:

It was found to be 0.99% Disintegration test:

It was found to be 2.8 minutes.

The study for ant diabetic activity was performed using chromogenic 3,5- dinitrosalicylic acid assay (DNSA method).

Table No. 5: Results of DNSA method by using colorimeter

*Syzygium cuminii*		*Gymnema sylvestre*		*Swertia chirata*
Dose (mg)	Response	Dose (mg)	Response	Dose (mg)	Response
100	0.60	100	0.53	100	0.21
150	0.65	150	0.55	150	0.29
200	0.71	200	0.56	200	0.31
300	0.79	300	0.65	300	0.37
350	0.80	350	0.68	350	0.41
400	0.82	400	0.72	400	0.43
450	0.80	450	0.73	450	0.45
500	0.83	500	0.76	500	0.46

From table No.5 it is clear that Syzygium cuminii has relatively potent alpha amylase inhibitory activity than Gymnema sylvestre and Swertia chirata.

· In this study we have prepared polyherbal tablet using these three medicinal plants based upon response mentioned in table No.5.

· Since we have prepared a 500mg tablet, amount of medicinal plant has been chosen from first three dosses only.

Graphs [dose (mg) versus response] of optimization study for medicinal plants Syzygium cuminii, Gymnema sylvestre, Swertia chirata

Figure no. 1: Dose Response Syzygium cuminii

Figure no. 2: Dose Response Gymnema sylvestre

Figure no. 3: Dose Response

Swertia chirata:

Table No. 6: Selected dose of medicinal plants.

Plant	Selected dose (mg)
Syzygium cuminii	100
Gymnema sylvestre	150
Swertia chirata	200

CONCLUSION:

In this study, the anti-diabetic activity of employed plant species Syzygium cuminii, Gymnema sylvestre and Swertia chirata was assessed using alpha amylase test and it was found to be effective. Also, the quality control tests of tablets were done according to the set of criteria of IP and the obtained results were found to be within the specified limits of each test. Further studies need to elucidate the molecular mechanism of interaction of plant-based drugs with human body in different disease

REFERENCES:

1. Kaul K, Tarr JM, Ahmad SI, Kohner EM, Chibber R. Introduction to diabetes mellitus. Diabetes: an old disease, a new insight. Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 771). 2013:1-1. doi: 10.1007/978-1-4614-5441-0_1.

2. G.S., Sharma, R., Sanodiya, B. S., Pandey, M., Prasad, G.B.K.S., Bisen P.S. Gymnema sylvestre: an alternative therapeutic agent for management of diabetes. Journal of Applied Pharmaceutical Science. 2012 Dec 29; 2(12): 1-6.doi:10.7324/JAPS.2012.21201.

3. Tiwari P, Mishra BN, Sangwan NS. Phytochemical and pharmacological properties of Gymnema sylvestre: an important medicinal plant. BioMed Research International. 2014 Jan https://:doi.org/10.1155/2014/830285.

4. Kumar V, Van Staden J. A review of Swertia chirayita (Gentianaceae) as a traditional medicinal plant. Frontiers in Pharmacology. 2016 Jan 12; 6: 308. https://doi/10.3389/fphar.2015.00308.

5. Chakravarty AK, Mukhopadhyay S, Das B. Swertane triterpenoids from Swertia chirata. Phytochemistry. 1991 Jan 1; 30(12): 4087-92. https://doi.org/10.1016/0031-9422(91)83473-X.

6. Lieberman HA, Lachman L, Joseph BS, Kanig JL. Preformulation. The Theory and Practice of Industrial Pharmacy; 3rd ed. Varghese Publishing House, Mumbai. 1987:183-4.

7. Shah MA, Khan RA, Ahmed M. Anti-diabetic activity of Iphiona aucheri leaf extract. Bangladesh Journal of Pharmacology. 2020 Oct 10; 15(4): 99-109.doi:10.3329/bjp.v15i4.44170.

Received on 10.05.2023 Modified on 26.10.2023

Res. J. Pharmacognosy and Phytochem. 2024; 16(2):69-72.

DOI: 10.52711/0975-4385.2024.00013