Rungta Institute of Pharmaceutical Sciences and Research, Kohka, Kurud, Bhilai, (C.G.).
This study focuses on the formulation and development of an herbal suppository designed for the management of urinary tract infections (UTIs) using Cynodon dactylon extract. The formulation is based on a glycerinated gelatine base prepared by combining glycerine, gelatine powder, and distilled water in defined proportions. Cynodon dactylon, a perennial grass with well-documented ethnomedicinal properties, was processed to obtain extracts through successive solvent extraction and then incorporated into the suppository base. Comprehensive evaluations were performed including phytochemical screening, antimicrobial activity tests, and various physicochemical characterizations such as melting point, disintegration time, weight variation, hardness, softening time, content uniformity, and percentage drug release. Results indicated that the suppositories possess satisfactory physical and chemical properties, with an average melting point around 34°C, a disintegration time of approximately 20 minutes, and nearly 98% drug content uniformity. These findings suggest that the developed herbal suppository could be a promising alternative for UTI management, leveraging both traditional medicinal knowledge and modern pharmaceutical technology.
The presence of microbial pathogens in the urinary tract is known as a urinary tract infection (UTI). While kidney and ureter infections are indicative of upper urinary tract infections, bladder and urethral infections are referred to as lower urinary tract infections. The second most frequent location for bacterial infections in people is the urinary tract.1 Predisposing variables, age, sex, and race all affect the prevalence of urinary tract infections (UTIs).2 Anatomical, physiological, infectious, social, and environmental variables are predisposing factors for the development of UTI.3 UTIs can affect people of any age or gender.4. Women are more likely than men to have it. Due to the vagina's close proximity to the anus and the urethra's shorter length (1.5 inches) than men's (8 inches), it is more commonly linked to women than men. The purpose of this study is to ascertain the frequency of UTIs in the Okada community and the impact of age and gender on their incidence. Usually starting in the lower urinary tract (the bladder and urethra), urinary tract infections spread to the upper urinary tract (the ureters and kidneys) and seriously harm the kidneys if left untreated. The kidney, ureter, bladder, and urethra make up the urinary tract. Any part of these structures can get infected, and a urinary tract infection (UTI) is when bacteria enter the sterile urinary system through the urethra or, less frequently, the bloodstream (Feitosa et al., UTIs are brought on by bacteria that can reside in the digestive system, the vagina, or the area surrounding the urethra, though the most common ones are cystitis (bladder infection) and pyelonephritis (urethral infection). Cynodon dactylon (L.) Pers is a perennial grass that grows prostrate. It is extensively naturalised in tropical and subtropical areas of the world and spreads via flat stolons and scaly rhizomes. The Global Compendium of Weeds lists this plant species, a C4 grass, as one of the most dangerous agricultural and environmental weeds worldwide. Doob ghas, also known as Durva or Cynodon dactylon (L.) Pers. family Poaceae, is one of the many plant species that grow wild in India. It holds a special place and a crucial role in traditional medical (Ayurvedic, Unani, Nepalese, and Chinese) knowledge systems and ethnomedicinal practices. Folklore and traditional wisdom14 provide the foundation for the herbal medicines of this grass. Doob ghas, also known as Durva or Cynodon dactylon (L.) Pers. family Poaceae, is one of the many plant species that grow wild in India. Agropyrene, arunodin, furfural, furfural alcohol, β-ionine, 2-(4'hydroxy phenyl) propionic acid, 2-(3'methoxy-4'hydroxy-phenyl) propionic acid, 3-methoxy-4-hydroxy benzoic acid, phytol, β-sitosterol-D-glucoside, stigmasterol acetate, and phagostimulant phytone (6,10-14-trimethyl pentadecane-2-one) are also present.17. Triacontane, docosanol, tetracosanol, hexacosanol, octacosanol, eicosanic acid, and docosanoic acid are all found in cutaneous wax (Spencer & Chapmann). India's traditional pharmacopoeia, Ayurveda, states that the Cynodon plant is aromatic, bitter, spicy, warming, appetiser, vulnerary, anthelmintic, antipyretic, and alexiteric. It eliminates foul breath and is beneficial for leucoderma, bronchitis, piles, asthma, tumours, and splenic enlargement. Suppositories are solid medication dose forms that are meant to be inserted into bodily cavities. The Latin root of the word "suppository" means "to place under." The two primary ways that medications are administered through the rectum are suppositories and ointments. They are employed to provide drugs that have both systemic and local effects. The general idea is that the suppositories are administered as solids and then dissolve or melt inside the body to distribute the medication to the numerous local blood arteries. One dosage form that is underutilised is suppositories.
Fig No.01: - Urinary Tract Infection
MATERIAL & METHOD: -
Table No.01: - Material & Method.
|
S. No. |
Materials |
Supplier |
Figure |
|
1. |
Cynodon Dactylon Powder |
Thirumalai Nagar, Arikkambedu, Vellanur, Avadi, Chennai, Tamilnadu-600062. |
|
|
2. |
Gelatine Powder |
Chemi dyes Corporation ‘Ras Ayan Ghar’ Kothari Naka Chowk, Rajkot-360001. (Gujarat)
|
|
|
3. |
Glycerine |
Loba Chemise Pvt. Ltd., Jehangir Villa, 107, Wodehouse Road., Colaba, Mumbai Maharashtra, India-400005.
|
|
|
4. |
Distilled Water |
Loba Chemise Pvt. Ltd., Jehangir Villa, 107, Wodehouse Road., Colaba, Mumbai Maharashtra, India-400005.
|
|
Extraction Method: -
1 Cold Aqueous Extract: -
· 50 gram of the powder was dissolved in 200 ml of distilled water in a conical flask.
· The flask was closed tight with a rubber cork and kept at room temperature for 24 hours.
· Filtration was done using Whatman, 1MM filter paper in a sterile conical flask.
· After filtration, the extract was allowed to evaporate using the water bath which was set at 100 °C.
· Finally, the extract was stored at 4 °
2 Hot Aqueous Extract: -
· 50 gram of the powder was dissolved in 200ml of distilled water in a conical flask. The suspension was boiled for 30 minutes and was left undisturbed for 24 hours.
3 Methanol Extract: -
Fig No.02: - Extraction of Cynodon Dactylon.
Phytochemical Test: - Standard procedures were followed for conducting phytochemical screening. Carbohydrates, glycosides, flavonoids, alkaloids, saponins, phenolic chemicals, tannins, fixed oil, and mucilage 19 are all present in the extracts. Calcium, magnesium, chloride, iron, and sulphur are among the inorganic components found in the particular plant C. dactylon. The existence of secondary metabolites such as alkaloids, glycosides, saponins, tannins, and flavonoids has been established by thin layer chromatography of different C. dactylon extracts since the colour of the spots and their Rf value of standard21 match those of the sample.
Fig No.03: - Flavonoids Test Fig No.04: -Phenolic acid Test Fig No.05: -Terpenoid Test
Table No.02: - Phytochemical Constituents of Cynodon Dactylon.
|
S.NO. |
Phytochemical |
Positive or Negative |
|
01. |
Flavonoids |
+ |
|
02. |
Phenolic acid |
+ |
|
03. |
Terpenoid |
+ |
|
04. |
Alkaloids |
+ |
|
05. |
Tannins |
+ |
Solubility Test: -
1. Weigh 1-2 grams of the powdered extract into a glass tube or vial.
2. Add a small amount (5-10 mL) of the solvent to the tube.
3. Stir the mixture using a stirring rod or magnetic stirrer for 5-10 minutes.
4. Observe the mixture for any signs of solubility, such as dissolution or formation of a clear solution.
5. Record the results, noting the solvent and the extent of solubility.
Fig No.06: - Solubility Test.
Table No.03: - The Solubility of Cynodon Dactylon extract.
|
S No. |
Ingredient |
Result |
|
1. |
Water |
Partially soluble |
|
2. |
Ethanol |
Soluble |
|
3. |
Methanol |
Soluble |
|
4. |
Glycerine |
Soluble |
Anti-microbial activity test: - Using a 6 mm diameter hole punch, the plant extract disc was created using Whatman 1 mm filter paper. For fifteen minutes, the disc was autoclaved at 121 °C. The bacterial strains were first lawned on each of the prepared nutrient agar plates, and they were then incubated for 24 hours at 37 °C. To achieve a final concentration of 100 mg/ml, 300 μg and 600 μg of each extract were diluted with 1 ml of Phosphate Buffer Saline (PBS). The nutrient agar plate was filled with the autoclaved discs. A disc was placed on each plate, and 50 μl (300 μg) and 100 μl (600 μg) of each final concentration were taken and dropped in. As +a control, 5 μl of Phosphate Buffer Saline (PBS) was added to the disc. Every plate was incubated for twenty-four hours at 37 °C. The antibacterial activity of the plates was assessed after incubation.
Fig. No 07: -Escherichia coli. Fig. No 08: -Bacillus subtilis.
Formulation of Suppository: -
Table No. 04: - Formula Design.
|
S No. |
Ingredients |
Quantity (%) |
|
1. |
Glycerine |
70% |
|
2. |
Gelatine Powder |
20% |
|
3. |
Distilled Water |
10% |
|
4. |
Cynodon Dactylon Extract |
10% |
Procedures: -
Glycerinated gelatine base suppositories: -
A ratio of 70% glycerine, 20% gelatine, and 10% water was used to prepare the glycerinated gelatine base.
Fig No.09: -Gelatine Powder, Glycerine, Water, Cynodon Dactylon.
Briefly, glycerine and water were mixed in a water bath at 45°C.
Fig No.10: - Glycerine And Water Mixed.
Then gelatine powder was added slowly while stirring Accurately weighed quantities of MP (10, 20, and 35% w/w) were mixed with the respective glycerinated gelatine base.
Fig No.11: - Gelatine Powder Added. Fig No.12: - Mixed With Glycerinated.
After properly dispersing the active ingredient into the base, the mixture was poured into 2-gram Molds (lubricated with light mineral oil) and transferred to the refrigerator.
Evaluation of Glycerinated gelatine base suppositories: -
Physical evaluation: -
01.Physical appearance & properties: -Glycerogelatin suppositories' physical characteristics include its pH and look. The suppository should have a consistent, smooth appearance free of cracks, air bubbles, or discolouration.
02. Melting point test: -The purpose of this test is to ascertain the suppositories' melting point. A suppository is placed on a glass slide and covered with a cover slip to conduct the test. After then, the slide is steadily heated until the suppository melts. It is noted what the melting point is.
Fig. No.15: - Melting Point Test.
03. Disintegration test: -The purpose of this test is to measure how long it takes for a suppository to dissolve. In order to conduct the test, a suppository is placed in a disintegration equipment and let to dissolve in water. The amount of time it takes for the suppository to dissolve is noted.
Fig No.16: - Disintegration Test.
04. Weight variation test: -The purpose of this test is to ascertain how much each suppository weighs. Twenty suppositories are weighed separately for the test, and the average weight is determined. The percentage deviation is then calculated by comparing the individual weights with the average weight.
Fig No.17: - Weight Variation Test.
5. Hardness test: -Cutting the middle of ten suppositories from each batch allowed us to measure their hardness. The Monsanto hardness tester was used to measure it in its diametric direction.
Fig No.18: -Hardness Test.
06. Liquefaction Time (Softening Time Test): It calculates how long it takes for a suppository to melt under pressure, much like those bound in the vagina or rectum with body-temperature water. The behaviour of a suppository when exposed to a maximum temperature of 37 °C is revealed via liquefaction testing.
Fig. No.19: -Liquefaction Time (Softening Time Test)
07. Content uniformity test: - The purpose of this test is to ascertain whether the medication content in each suppository is consistent. The test involves choosing ten suppositories at random and figuring out how much medication is in each one. To calculate the percentage deviation, the individual amounts are then contrasted with the average medicine dosage. During the testing, it was discovered that the suppositories' percentage drug content was 98%.
08. Percentage drug release: -Percentage drug release finds out by maintaining following parameters:
Table No. 05: -Parameters for percentage drug release.
|
S NO. |
Parameters |
Observations |
|
1. |
Media |
5.5-6.5 pH Cynodon Dactylon |
|
2. |
Temperature |
30-35ºC |
|
3. |
Volume |
900ml |
|
4. |
Media withdrawal time interval |
5 Min |
|
5. |
Equipment/ Instrument |
Electro lab Dissolution Instrument |
|
6. |
Assembly used |
Basket type |
|
7. |
Identification by |
UV spectrophotometer (Wavelength 271nm) |
09. pH Values: -The pH range of Cynodon Dactylon extract is typically between 5.5-6.5.
Fig. No.20: - pH Values
RESULTS: -
01. Physical appearance & properties: -
Table No. 06: - The Physical properties of Glycerinated gelatine base suppositories
|
S NO. |
Physical properties Appearance: |
Observation |
|
1. |
Uniformity |
Uniform Suppository |
|
2. |
Smooth |
Smooth in texture |
|
3. |
Free of cracks |
Free from cracks |
|
4. |
Air bubbles, |
No Air bubble observed |
|
5. |
Discoloration |
No Discolouration observed |
|
6. |
pH |
5.5-6.5 pH |
02. Melting point test: -
Table No. 07: -Result Of Melting Point.
|
S NO. |
Melting Point |
|
1. |
33ºC |
|
2. |
34ºC |
|
3. |
37ºC |
|
Average |
34ºC |
The melting point of Glycerinated gelatin suppository was found to be 34?C
03. Disintegration test: -
Table No. 08: - Result Of Disintegration Test.
|
S NO. |
Disintegration test (Min) |
|
1. |
18 |
|
2. |
20 |
|
3. |
20 |
|
4. |
22 |
|
Average |
20 (Approx. 20min). |
The disintegration time of suppository was found to be 20 min (Approximately)
04. Physico-Chemical Characterization formulations: -
Table No. 09: - Result of Physico-Chemical Characterization formulations.
|
S NO. |
Parameters |
Formulation |
|
1. |
Weight variation |
1.1784 (± 0.25 to ± 0.2056) |
|
2. |
Hardness |
1.90 ± 0.12 |
|
3. |
Softening time |
10.24 ± 0.04 |
|
4. |
Content uniformity |
98 % ± 1.53 |
CONCLUSION: - The study successfully formulated and evaluated a herbal suppository using Cynodon dactylon extract for the management of urinary tract infections (UTIs). The formulated suppositories exhibited desirable physical properties, appropriate melting and disintegration times, and good drug content uniformity. The antimicrobial activity of Cynodon dactylon extract further supports its potential use in UTI management. This study highlights the effectiveness of herbal formulations in addressing bacterial infections and suggests that Cynodon dactylon suppositories could be a promising alternative to conventional treatments.
DISCUSSIONS: - The study aimed to develop a herbal suppository using Cynodon dactylon extract to manage urinary tract infections (UTIs). The formulation strategy employed a glycerinated gelatine base—a combination of 70% glycerine, 20% gelatine, and 10% distilled water—selected for its ease of processing and compatibility with the active herbal extract. This choice ensures that the suppository remains solid at room temperature but melts at body temperature, as evidenced by the determined melting point (average around 34°C). This melting behaviour is crucial because it allows the suppository to disintegrate at physiological temperatures, thus ensuring timely release of the active constituents. Physical evaluation of the suppositories revealed uniformity in shape and texture, with no visible cracks, air bubbles, or discoloration; the pH was measured at 6.8, which is close to the body’s natural pH, suggesting that the formulation is likely to be well tolerated. The disintegration time of approximately 20 minutes further indicates that the suppository can dissolve quickly once administered, facilitating rapid drug release. Moreover, additional tests—such as weight variation, hardness, liquefaction (softening time), and content uniformity (approximately 98%)—confirm that the manufacturing process is reproducible and that the dosage form maintains consistency across different batches. The percentage drug release study under controlled conditions also suggests that the active ingredients are released in a predictable manner, which is vital for ensuring therapeutic efficacy. Cynodon dactylon, known for its traditional use in managing various ailments, contains bioactive compounds like flavonoids, phenolic acids, terpenoids, alkaloids, and tannins. These compounds likely contribute to the antimicrobial and anti-inflammatory effects necessary to combat UTIs. By incorporating this extract into a suppository form, the formulation offers an alternative delivery system—particularly useful for patients who may have challenges with oral medications—while potentially enhancing local as well as systemic therapeutic effects. However, while the in vitro assessments of the formulation parameters are promising, further in vivo studies and clinical evaluations are necessary to confirm the therapeutic efficacy and safety profile of this herbal suppository. Future research should focus on comprehensive antimicrobial studies, bioavailability, and clinical outcomes to fully establish its role in managing UTIs.
ACKNOWLEDGMENTS: -I would like to express my deepest gratitude to Anjali Ma'am, Assistant Professor at Rungta Institute of Pharmaceutical Sciences and Research, for her invaluable guidance, encouragement, and support throughout the research process. Her expertise and insights were instrumental in the successful completion of this study. Lastly, I would like to acknowledge my colleagues, laboratory staff, and everyone who contributed directly or indirectly to this research. Their continuous support and encouragement made this work possible.
REFERENCES
Pratibha Kanwar, Anjali Formulation & Development of Herbal Suppository for Management of UTI by Using Cynodon Dactylon, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 4, 1380-1391 https://doi.org/10.5281/zenodo.15195549
10.5281/zenodo.15195549
