Department of Pharmaceutics, Govindrao Nikam College of Pharmacy, Sawarde, Ratnagiri, India
Diabetes mellitus is a condition that can have long-term complications. This study provides information on herbal medications that can treat a patient's diabetes complications. Herbal anti-diabetic medications reduce blood glucose levels and stimulate pancreatic secretion of insulin to treat diabetes mellitus. According to this research, durva and neem mostly exhibit antidiabetic properties because of various chemical component that lowers blood glucose levels. Neem acts as a wound healer and has anti-hyperglycemic properties when a diabetic patient sustains any kind of injury. In this study, a natural anti-diabetic medication was created using neem and durva, and various tests were conducted to assess its effectiveness, including the hardness, friability, weight variation, and dissolution tests.
Diabetes Mellitus:
It is characterized by elevated blood glucose levels. The primary classifications include Type 1 and Type 2 diabetes. However, additional forms exist, such as gestational diabetes, neonatal diabetes, maturity-onset diabetes of the young (MODY), and diabetes resulting from secondary causes, including other medical conditions or steroid therapy. Type 2 diabetes is typically associated with insulin resistance, predominantly affecting older adults, and is frequently linked to low-fiber diets and specific lifestyle factors. In contrast, Type 1 diabetes results from insufficient insulin production and most commonly presents in children or adolescents.(1)
Type 1 diabetes (β-cell destruction, usually leading to absolute insulin deficiency)
Diabetes, a type of type 1 diabetes, is caused by the immune system's destruction of pancreatic β-cells, resulting in autoantibodies against islet cells, affront, GAD65, and IA-2 and IA-2β. This type affects 5-10% of people with diabetes.
Type 2 diabetes (ranging from predominantly insulin resistance with relative insulin deficiency to predominantly an insulin secretory defect with insulin resistance)
Non-insulin-dependent diabetes, also known as type 2 diabetes, affects 90-95% of individuals. These individuals have relative insulin shortage and are insulin resistant, often surviving without medication. The exact causes remain unknown, and they do not have autoimmune destruction of β-cells.(2)
Durva (Cynodon Dactylon)
Cynodon Dactylon is a hardy, perennial grass found in India, growing up to eight thousand feet above sea level. It has long runners, or stolons, that root at nodes and form a dense tuff on the soil's surface. The inflorescence of clums ranges in height from 15 cm to 1 m and is made up of 2-12 spikes arranged star-shaped at the tip of the stem. The grass is common along trails and roadside edges, encroaching on uncultivated land quickly. It blooms for almost the whole year and is vital for conservation due to its ability to stop soil erosion. Cynodon dactylon is utilized in traditional medicine for wound healing and the treatment of dyspepsia. During fermentation, it imparts a sour flavor to beer. The plant exhibits multiple health-related properties, including vulnerary, alterative, antiseptic, astringent, cyanogenetic, demulcent, depurative, diuretic, emollient, sudorific, and photosensitizing effects in animals. Additionally, it has been associated with hay fever and contact dermatitis.(3)
Neem (Azadirachta indica)
The Meliaceae family tree Azadirachta indica A. Juss, commonly referred to as neem, is used medicinally to treat conditions like intestinal helminthiasis, skin infections, and respiratory infections. It has various organic and pharmacological activities, including antiviral, antibacterial, anti-inflammatory, antipyretic, and antioxidant properties. The wood is also used for timber, reforestation, Shadean animal and poultry feed, fertilizers, and pesticides. It grows well in many arid regions with annual rainfall ranging from 400 to 1299 mm. However, it does not tolerate ice or moderate cold. The Azadirachta indica tree has adapted to the unique climate and soil types of South Korea and strives to grow it in a warm and peaceful climate.(4)
Antidiabetic Activity Principle
In vitro measurements of alpha amylase activity can be made by hydrolyzing starch when the enzyme is present. The DNSA reagent can be used to quantify this process since it reacts with the reducing sugar maltose (a hydrolyzed product of starch) to produce a red color. The degree of redness signifies the breakdown of starch into maltose caused by enzymes. If the extract has alpha-amylase inhibitory activity, the red color will be less intense. That is, the alpha-amylase inhibitory activity is inversely related to the intensity of the red color.
Determining the antidiabetic activity of the Cynodon Dactylon. Formulation and Evaluation of Poly Herbal Antidiabetic Tablet. Develop a polyherbal tablet formulation using Durva (Cynodon Dactylon) and Neem (Azadirachta indica) extracts, for their anti-diabetic activity. Optimize the composition of the herbal blend to achieve maximum potency in lowering blood glucose levels. Perform an anti-diabetic assay of the selected extracts of Durva (Cynodon Dactylon) and Neem (Azadirachta indica) extracts. Formulate tablet dosage forms using selected herbal extracts and excipients, employing a wet granulation method. Perform the evaluation studies of the formulated tablets, including weight variation, hardness, thickness, and friability, adhering to pharmacopeial standards for quality assurance.(5)
TABLE 1: PHARMACOLOGICAL ACTIVITY OF CYNODON DACTYLON
|
Parts / Extracts |
Biological action |
|
Aqueous |
Anti-Diabetic |
|
Ethanolic |
Anti-Diabetic |
|
Hydroalcoholic solution |
Cardio Protective |
|
Aq. solution |
Antioxidant |
|
Methanolic |
Anti-Diabetic |
|
50% Aqueous-Ethanolic Extract |
Nephrolithiasis (Kidney stone Removal) |
|
Shoot |
Immunomodulatory & DNA Protective |
TABLE 2: CHEMICAL CONSTITUENT
|
Chemical constituent |
Activity |
|
Flavone |
Antidiabetic |
|
B-sitosterol, - carotene |
Antidiabetic |
|
phytol |
Anticancer, Antimicrobial |
|
Diazoprogesterone |
Antimicrobial |
|
Glycerin |
Anti-inflammatory, Antimicrobial |
|
Benzenepropanol, |
Antioxidant |
MATERIAL AND METHODS:
Apparatus: Test tubes, test tube holder, beaker, stirrer, water bath, incubator, pipette, etc.
Chemicals: DNSA reagent, Alpha amylase enzyme, starch paste, phosphate buffer, extract, etc.
PREPARATION OF EXTRACT:
Methanolic extract of Cynodon Dactylon:
The Cynodon Dactylon plant was the source of the plant material. The gathered material should be cleaned and dried. The plant material was ground into a fine, dry powder once it had dried. In a beaker, 100 milliliters of methanol were used to dissolve 15 grams of the powder. For around twenty-four hours, the solution was kept at 25°C and 50 rpm on a rotary shaker. Whatman 1mm filter paper was used for filtration in a sterile conical flask. The extract was filtered and then left to dry at room temperature. Lastly, 4°C was used to preserve the extract.
Cold aqueous extract of Cynodon Dactylon:
In a conical flask, 50 grams of the powder were dissolved in 200 milliliters of distilled water. The flask was kept on a rotary shaker at room temperature for a full day after being securely sealed with a rubber cork. Whatman 1MM filter paper was then used for filtering in a sterile conical flask. Following filtration, a water bath set at 100 °C was used to let the extract evaporate. Lastly, 4°C was used to preserve the extract.
Anti-diabetic assay procedure:
Take 3 test tubes, the first test tube contains a blank solution that is the DNSA reagent, and the solution of alpha amylase enzyme. In the second test tube, a solution of extract, DNSA reagent, starch paste, and alpha amylase solution is added. The third test tube contains DNSA reagent, starch paste, and alpha amylase enzyme. In addition to this, observe the color intensity of the test tubes second and third. Take different quantities of extract in 5 test tubes at a concentration of 500 μg/ml. (0.2, 0.4, 0.6, 0.8,1 ml). And 2 test tubes, one without starch and the other one with starch. Add enzyme and buffer to all test tubes. Make sure to shake them properly. Ten minutes are spent incubating the test tubes at room temperature. Next, fill each test tube with 1 milliliter of 1% starch paste. Once more, let the reaction sit at room temperature for roughly 20 minutes.
To each test tube, add one milliliter of DNSA reagent. For approximately fifteen minutes, incubate the test tube in a boiling water bath until the red color develops. After that, measure the absorbance at 545 nm. Examine the enzyme activity of blank, inhibitor-free, and inhibitor-containing samples at various concentrations.(5)
From the above readings, the enzyme activity is calculated.
Enzyme activity =
Release μg of maltose x 100 Molecular weight of maltose x incubation time
Where,
μg of maltose released = Absorbance / slope
Molecular weight of maltose = 342 g/mol
Incubation time = 10 minutes
TABLE 3: ABSORBANCE AT DIFFERENT CONCENTRATIONS
|
Enzyme (0.5 %) (ml) |
Buffer (pH 7) (ml) |
Inhibitor (500 μg/ml) |
Starch (1%) (ml) |
DNSA reagent (ml) |
Absorbance |
|
0.5 |
1 |
- |
- |
1 |
1.331 |
|
0.5 |
1 |
- |
1 |
1 |
2.332 |
|
0.5 |
0.8 |
0.2 |
1 |
1 |
1.466 |
|
0.5 |
0.6 |
0.4 |
1 |
1 |
1.164 |
|
0.5 |
0.4 |
0.6 |
1 |
1 |
0.685 |
|
0.5 |
0.2 |
0.8 |
1 |
1 |
0.457 |
|
0.5 |
- |
1 |
1 |
1 |
0.257 |
TABLE 4: ENZYME ACTIVITY OF CYNODON DACTYLON AT DIFFERENT CONCENTRATIONS
|
Test |
Concentration |
Enzyme activity (μmol/min) |
|
Without extract |
- |
0.454 |
|
With the methanolic extract
|
100 (μg/ml) |
0.285 |
|
200 (μg/ml) |
0.226 |
|
|
300 (μg/ml) |
0.133 |
|
|
400 (μg/ml) |
0.089 |
|
|
500 (μg/ml) |
0.050 |
TABLE 5: ENZYME ACTIVITY OF AZADIRACHTA INDICA AT DIFFERENT CONCENTRATIONS
|
Test |
Concentration |
Enzyme activity (μmol/min) |
|
Without extract |
- |
0.612 |
|
With extract
|
100 (μg/ml) |
0.402 |
|
200 (μg/ml) |
0.290 |
|
|
300 (μg/ml) |
0.285 |
|
|
400 (μg/ml) |
0.235 |
|
|
500 (μg/ml) |
0.179 |
From the above calculations, it has been observed that the selected herbs Cynodon Dactylon and Azadirachta indica are potent at 200(μg/ml).
Fig. 1: Cynodon Dactylon extract Fig. 2: Azadirachta indica extract
We can see from the test tubes above that test tube number three has a lower color intensity than test tube number two. The enzyme-induced hydrolysis of starch into maltose is shown by the intensity of red color in test tube number two. Alpha-amylase enzyme inhibitory action is demonstrated by the extract of Azadirachta indica and Cynodon dactylon.
Wet granulation:
Weigh and sieve the extracts. Mix them with HPMC polymer. Add a 5% solution of PVP binder with isopropyl alcohol solvent to the mixture. Prepare a damp mass of the mixture. Dry the mixture in an oven at 50 °C for about 10-15 minutes. After drying, sieve the mixture using a 14/18 mesh sieve. Dry the granules again and pass them through a No. 22 sieve. Add up to 15% fines to the granules and conduct pre-granulation studies. Add postgranulating materials, i.e., magnesium stearate and talc. Perform post-granulation studies.(6)
TABLE 6: FORMULATION TABLE
|
Sr No |
Ingredient |
Quantity (10 Tablets) |
Role |
|
1) |
Neem |
2g |
API |
|
2) |
Durva |
2g |
API |
|
3) |
HPMC |
0.7g |
Polymer |
|
4) |
Magnesium Stearate |
0.2g |
Glidant |
|
5) |
Talc |
0.1g |
Lubricant |
|
6) |
PVP (5% solution in IPA) |
q.s. |
Binder / Diluent |
Fig. 3: Granules
Pre-formulation studies:
Angle of repose: The maximum angle that may be made between the surface of a pile of powder and a horizontal plane is called the angle of repose, and it signifies insufficient drug flow. It rises when particle surfaces are uneven or rough.
Bulk density: It, which may be written as g/cm3, kg/m3, or g/100 ml, is the weight of a powder per unit volume. Standardizing tapping and measuring the amount of 100 grams of powder in a 250 ml graduated cylinder, following compaction, are the methods used to compute it. With this approach, precise measurements and computations for powders are guaranteed.
Tapped density: Tapped density is a higher bulk density that is achieved by automatically tapping a graduated measuring cylinder or vessel, or by mechanically tapping a container holding a powder sample.
Hausner’s ratio: For the purpose of identifying powder flow properties and interparticle friction, Hausner's ratio is essential. The ratio is around 1.2 for low-friction powders, such as coarse spheres, and more than 1.6 for cohesive, less free-flowing powders.
Carr’s compressibility index: Particle size, cohesiveness, and relative flow rate of powder all have a direct bearing on it. It is a well-known, quick, and straightforward method of reigning over powder glide characters. A unit named percentage is used to express the Carr's index.(6)
Post formulation studies:
The addition of post-granulating material, such as glidant and lubricant is done, and then the post-formulation study was performed. After these studies, the compression was done using a single rotary compression machine.
Compression of tablet: Compress the granules using a single rotary compression machine.
EVALUATION TEST:
Weight variation test: The weight variation test (WFD) would be a good way to determine the homogeneity of drug residue in drug distribution. In exercise, 20 tablets are taken from a batch. The 20 tablets are weighed. The average weight is taken from the 20 tablets. Then the tablets are weighed individually.
Hardness test: A tablet's crushing strength is another name for its hardness test. The force needed to shatter the tablet is measured by this test. To determine the tablet's hardness, the Monsanto hardness tester is utilized. The unit of measurement for hardness is kg/cm3.
Friability test: The Roche Friabilator is a laboratory friability test used to evaluate the abrasion and the stock effect on antidiabetic tablets. It involves spinning a pre-weighed sample, weighing, and dusting, ensuring appropriate transit.
Dissolution test: The time it takes for a tablet to shatter under test circumstances is a measure of dissolution, which is the process by which a solid solute dissolves into a solution. Since acidic medications are absorbed quickly, the dissolution rate was measured by spinning 900 milliliters of 0.1N HCL for 120 minutes.(7)
RESULT AND DISCUSSION:
From the above readings, it has been observed that the selected herbs Cynodon Dactylon and Azadirachta indica are potent at 200(μg/ml). In order to create dosage forms that are stable, effective, and safe for end users, innovators and industry professionals must first conduct pre-formulation studies. Pre-formulation is a critical concept for the development of final formulation dosage forms that are intended to target a disease. This is why pre-formulation studies are conducted; they yield useful data and reveal important information that can be used to produce dosage forms that are safe, effective, and stable for end users. From the above table, it is concluded that the pre-formulation studies conducted show poor flow properties. Therefore, we added the glidant and lubricant into the granules and again conducted the same test as post-formulation studies. From the post formulation studies it is concluded that the flow properties of the granules are excellent, so that we can use the granules for the tablet compression. Therefore, we concluded that the pre-formulation studies enhance guidance, regulatory relief, resource conservation, public safety standards, and product quality in drug development and evaluation, while conserving resources.
CONCLUSION
The patient's cure or treatment for diabetes was the primary goal of the herbal anti-diabetic tablet. The research came to the conclusion that herbal anti-diabetic tablets made from natural sources have fewer negative effects than tablets made from synthetic compounds. Using a variety of metrics, the prepared herbal anti-diabetic tablet was proven to be effective in treating diabetic patients.
REFERENCES
Pomaje M. D., Dr. Patil A. B., Gangan S. M., Acharya A. A., Nantekar S. S., Formulation Development and Evaluation of Polyherbal Formulation for the Management of Type-2 Diabetes, Int. J. of Pharm. Sci., 2026, Vol 4, Issue 2, 3076-3082. https://doi.org/10.5281/zenodo.18700466
10.5281/zenodo.18700466
