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Padmini College of pharmacy, Dighanchi, Atpadi, Sangli
Background: Diuretic agents are commonly prescribed for conditions such as hypertension, edema, and renal disorders due to their ability to increase urinary excretion of water and electrolytes. Continuous use of synthetic diuretics may produce adverse effects including electrolyte imbalance, creating interest in plant-based alternatives. Objective: The present investigation was carried out to assess the in-vitro diuretic activity of aqueous extract of Cynodon dactylon by mercurimetric estimation of chloride ions using acetazolamide as a reference standard. Materials and Methods: The collected plant material of Cynodon dactylon was shade dried, powdered, and extracted with distilled water. Preliminary phytochemical screening was performed using standard qualitative procedures. Diuretic activity was evaluated by mercurimetric titration, in which chloride ions were estimated using standard mercuric nitrate solution and diphenylcarbazone indicator. The chloride ion concentration of control, test, and standard samples was compared. Results: Phytochemical analysis confirmed the presence of flavonoids, alkaloids, tannins, glycosides, and saponins in the aqueous extract. The control sample exhibited chloride concentration of 58 mEq/L, while the plant extract showed reduced chloride concentration of 28.2 mEq/L corresponding to 51% reduction. Acetazolamide demonstrated maximum reduction with chloride concentration of 8.2 mEq/L and 85% reduction. The findings indicate notable diuretic activity of the plant extract. Conclusion: The aqueous extract of Cynodon dactylon showed significant in-vitro diuretic potential by enhancing chloride ion excretion. The study supports the traditional medicinal use of the plant as a natural diuretic and suggests the need for further pharmacological and clinical investigations.
Diuretic therapies play a crucial role in the management of cardiovascular and renal disorders by promoting the elimination of excess fluids and electrolytes from the body¹. Although synthetic diuretics are clinically effective, their prolonged use may lead to metabolic imbalances and electrolyte disturbances, thereby necessitating the development of safer alternatives²
Diuretics Administration
↓
Act on Nephron (Different Sites)
↓
↓ Sodium (Na⁺) Reabsorption
↓
↑ Sodium in Tubular Fluid
↓
Water Follows (Osmosis)
↓
↑ Urine Output (Diuresis)
↓
↓ Blood Volume
↓
↓ Cardiac Output
↓
↓ Blood Pressure
Diuretics work by acting on the nephron in the kidney and reducing the reabsorption of sodium. As a result, more sodium remains in the filtrate, and water moves along with it due to osmotic effects, leading to increased urine formation. The loss of fluid decreases the overall blood volume, which in turn reduces cardiac output and helps lower blood pressure. This action can also lead to imbalances in electrolytes, such as decreased or increased potassium levels. Additionally, it may disturb the body’s acid–base balance and trigger compensatory mechanisms like activation of the renin–angiotensin–aldosterone system (RAAS).
Cynodon dactylon (L.) Pers., a widely distributed perennial grass belonging to the Poaceae family, has emerged as a promising natural candidate³. It is commonly known as Bermuda grass, Doob, or Durva and is abundantly found in tropical and subtropical regions⁴. The botanical nomenclature reflects its morphological features, with the term Cynodon derived from Greek meaning “dog tooth,” and dactylon indicating its finger-like inflorescence⁵. In India, Cynodon dactylon holds significant cultural and medicinal importance. It is considered sacred and is traditionally offered to deities such as Shiva, Ganesha, and Vishnu. Additionally, it is an important component of the Ayurvedic formulation ‘Dasapushpam’⁶. Traditionally, the plant has been used in indigenous medicine for the treatment of dysentery, urinary disorders, and as a natural diuretic agent⁷. The therapeutic potential of Cynodon dactylon is attributed to its rich phytochemical composition, which includes flavonoids, saponins, triterpenoids, alkaloids, and phenolic compounds⁸. These bioactive constituents are responsible for various pharmacological activities such as anti-inflammatory, antioxidant, antimicrobial, and diuretic effects⁹. Therefore, the present study aims to evaluate the pharmacological potential of Cynodon dactylon, with particular emphasis on validating its traditional use as a natural diuretic agent and exploring its future therapeutic applications
Standard Drug (Acetazolamide):
Acetazolamide is a carbonic anhydrase inhibitor that reduces the reabsorption of sodium bicarbonate in the proximal tubule of the kidney, leading to increased bicarbonate excretion in urine. Because of this effect, it is commonly used in the treatment of metabolic alkalosis. Carbonic anhydrase normally helps in hydrogen ion secretion and bicarbonate reabsorption in the kidneys, as well as carbon dioxide transport in the body. By inhibiting this enzyme, acetazolamide causes alkaline diuresis and increases the elimination of bicarbonate in urine. Acetazolamide is a sulfonamide derivative that acts as a non-competitive inhibitor of carbonic anhydrase throughout the body. It is usually administered orally, although intravenous administration is also possible. Oral bioavailability is about 90%, and most of the drug binds to plasma albumin, allowing wide distribution including the brain. The onset of action occurs within about 1 hour, with peak effects at 2–4 hours. The drug has a half-life of 10–15 hours and is primarily eliminated unchanged through the kidneys.
MECHANISM OF ACTION
Acetazolamide inhibits the enzyme carbonic anhydrase in the proximal renal tubule. This reduces hydrogen ion formation and decreases the Na⁺/H⁺ exchange process. As a result, the reabsorption of sodium and bicarbonate decreases, causing more sodium, bicarbonate, and water to remain in the tubular lumen. This leads to increased urinary excretion and diuresis.
Drug inhibits Carbonic anhydrase (CA)
↓
Reduced conversion of carbonic acid in the proximal tubule
↓
↓H⁺ formation inside tubular cells →less Na⁺/H⁺ exchange
↓
↓Na⁺ and HCO3⁻re-absorption →more Na⁺, HCO3 and water stay in the lumen
↓
Increased urinary excretion of Na⁺, HCO3⁻, water → dieresis
MATERIALS AND METHODS
Collection and Authentication of Plant Material
Fresh whole plant material of Cynodon dactylon was collected from the local area of Dighanchi, Maharashtra, India. The plant was authenticated by the Department of Botany, Ishwarrao More Patil Arts, Commerce and Science Mahila Mahavidyalaya, Dighanchi. The collected material was washed thoroughly with tap water to remove adhering dust and impurities, shade dried room temperature, and pulverized into coarse powder using a mechanical grinder. The powdered material was stored in an airtight container for further study.
Fig No 1. Photograph Showing Cynodon Dactylon
Preparation of Aqueous Extract
About 25 g of powdered plant material was mixed with 250 mL of distilled water in a suitable container. The mixture was allowed to stand for 30 min followed by heating on a water bath for another 30 min with occasional stirring. The extract was cooled to room temperature and filtered initially through muslin cloth and then through Whatman filter paper to obtain a clear filtrate. The prepared aqueous extract was stored at 4°C until use.
Fig No 2. Prepared Plant Extract
Preliminary Photochemical Screening
The aqueous extract of Cynodon dactylon was subjected to preliminary phytochemical screening using standard qualitative methods for the detection of alkaloids, flavonoids, glycosides, tannins, and saponins. Formation of characteristic color changes or precipitates was considered as positive indication for the respective phytoconstituents.
|
Sr No. |
Test |
Inference |
|
1) |
Flavonoids |
Present |
|
2) |
Alkaloids |
Present |
|
3) |
Tannis |
Present |
|
4) |
Saponins |
Present |
Fig No 3. Phytochemical Screnning Test
Principle of Mercurimetric Method
The mercurimetric titration method is based on the reaction between chloride ions and mercuric nitrate to form mercuric chloride. After complete reaction of chloride ions, excess mercuric ions react with diphenylcarbazone indicator producing a violet-blue colored complex, indicating the end point of titration.
Reagents and Chemicals
The following reagents were used for chloride ion estimation:
All chemicals used were of analytical grade.
Preparation of Control, Test, and Standard Solutions
Control Solution
A chloride-containing control solution was prepared using sodium chloride solution.
Test Solution
The test solution was prepared by mixing a measured quantity of aqueous extract of Cynodon dactylon with the chloride-containing solution.
Standard Solution
Acetazolamide solution was prepared and used as standard diuretic drug.
Estimation of Chloride Ions
For chloride ion estimation, 10 mL of each sample solution was transferred separately into a conical flask. To this, 1 mL nitric acid and 1 mL diphenylcarbazone indicator were added. The solution was titrated against standardized mercuric nitrate solution until the appearance of violet-blue end point.
The experiment was carried out for:
All observations were recorded in triplicate.
Fig No 4. Estimation Of Chloride Ion In Standard Drug
Calculation of Chloride Ion Concentration
The chloride ion concentration was calculated using the following formula:
Chloride concentration (mEq/L)=Sample volumeV×N×1000
Where:
Statistical Analysis
All experiments were performed in triplicate (n = 3) and results were expressed as Mean ± Standard Deviation (SD). Statistical significance was determined by comparing the test and standard groups with control values, and a value of p < 0.05 was considered statistically significant.
|
Sample |
Reading 1 (mL) |
Reading 2 (mL) |
Reading 3 (mL) |
Mean ± SD |
|
Control |
29 |
28 |
30 |
29.0 ± 1.0 |
|
Test Extract |
13 |
15 |
14.5 |
14.1 ± 1.04 |
|
Standard (Acetazolamide) |
3 |
4 |
5.5 |
4.1 ± 1.26 |
|
Sample |
Average Volume(V) |
Chloride Concentration (mEq/L) |
|
Control |
29 |
58 |
|
Test(Extract) |
14.1 |
28.2 |
|
Standard(Acetazolamide) |
4.1 |
8.2 |
|
Sample |
% Reduction |
|
Test(Extract) |
51% |
|
Standard(Acetazolamide) |
85% |
Interpretation of Results:
DISCUSSION:
The present study demonstrates the in-vitro diuretic activity of Cynodon dactylon using Mercurimetric estimation of chloride ions. The method is based on the reaction between chloride ions and mercuric nitrate, forming mercuric chloride, with endpoint detection by diphenylcarbazone indicator. The control sample showed the highest chloride concentration (58 mEq/L), whereas the test extract showed reduced chloride concentration (28.2 mEq/L), indicating moderate diuretic activity. The standard drug Acetazolamide exhibited maximum reduction (85%), confirming its strong diuretic effect. The observed activity of Cynodon dactylon may be attributed to phytoconstituents such as flavonoids, saponins, alkaloids, and tannins, which are known to influence renal function and electrolyte balance. Although the extract showed less activity compared to the standard drug, it demonstrated significant diuretic potential and may serve as a natural alternative, warranting further in-vivo and clinical studies.
CONCLUSION
The findings of the present study indicate that the aqueous extract of Cynodon dactylon possesses considerable in-vitro diuretic activity as evidenced by reduced chloride ion concentration in comparison with the control sample. The Mercurimetric chloride estimation method demonstrated that the plant extract produced moderate diuretic effect, although lower than the standard drug Acetazolamide. The observed activity may be associated with the presence of phytoconstituents such as flavonoids, saponins, alkaloids, and tannins. These results provide scientific support for the traditional use of Cynodon dactylon in urinary and renal disorders. Further in-vivo studies and detailed pharmacological investigations are necessary to confirm its efficacy and safety for therapeutic applications.
ACKNOWLEDGEMENTS
We express our sincere gratitude to Padmini College of Pharmacy, Dighanchi, for providing the necessary facilities and support to carry out this research work. We are deeply thankful to our respected guide and faculty members for their valuable guidance, constant encouragement, and constructive suggestions throughout the course of this study. Their support has been instrumental in the successful completion of this work. We also extend our thanks to the teaching and non-teaching staff of the college for their cooperation and assistance during the experimental work. Finally, we express our heartfelt gratitude to our friends and family members for their continuous motivation and support.
REFERENCES
Neha Mane, Vaishnavi Mane, Shweta More, Saragam Masugade, Aakash Misal*, Amit Panaskar, Bhagyashri Panaskar, Assessment of Diuretic Potential of Cynodon Dactylon Through Chloride Ion Estimation by Mercurimetric Method: An In-Vitro Study, Int. J. Med. Pharm. Sci., 2026, 2 (5), 402-408. https://doi.org/10.5281/zenodo.20093164
10.5281/zenodo.20093164