FORMULATION AND EVALUATION OF A NOVEL POLYHERBAL ORAL MEDICATED JELLY FOR TREATMENT OF UROLITHIASIS

FORMULATION AND EVALUATION OF A NOVEL POLYHERBAL ORAL MEDICATED JELLY FOR TREATMENT OF UROLITHIASIS

Soham S. Mule, Harsh A. Naik, Khushal R. Solanki, Lokesh Devnani and Mrinal Sanaye *

Department of Pharmacology, Principal K. M. Kundnani College of Pharmacy, Mumbai, Maharashtra, India.

ABSTRACT: Background: Urolithiasis is a disease of the genitourinary system, characterised by occurrence of urinary stones and affecting approximately 12% of the global population. It is also linked to chronic diseases like chronic renal disease, cardiovascular disease thereby posing significant health risks. Contemporary treatments are symptomatic and fail to produce lithotriptic activity and thus recurrence is common. This study involves development of polyherbal formulation and evaluation of its lithotriptic activity. Methodology: Hydroalcoholic (HA) extracts of Tribulus terrestris, Cynodon dactylon, and Kalanchoe pinnata were formulated into a polyherbal oral medicated jelly (OMJ). The OMJ was evaluated for characteristics like hardness, cohesion, springiness, gumminess, and chewiness using a texture analyser. The lithotriptic activity was evaluated using HPLC analysis. Calcium oxalate crystals were incubated with HA extracts in egg membrane for 72 hours. The residual contents were then analysed using HPLC. Cystone was used as a standard. Results: Texture analysis of OMJ revealed the characteristics such as: gumminess (431g), springiness (36.70), hardness (518g), cohesiveness (0.83), and chewiness (155 mJ) at pH of 4.3. The dissolution of calcium oxalate in presence of OMJ was found to be 73.9%, whereas dissolution of 92.5% was observed with Cystone. Conclusion: The OMJ was developed and its in-vitro lithotriptic effect was evaluated which demonstrated the potential therapeutic efficacy of OMJ in dissolving calcium oxalate crystals thereby envisaging the role of OMJ in management of urolithiasis. Further preclinical investigations are needed to validate its therapeutic efficacy and safety.

Keywords: Lithotriptic activity, Oral medicated jelly, Polyherbal formulation, Urolithiasis

INTRODUCTION: Urolithiasis is a renal complication associated with occurrence of stones across the urinary tract ¹. Commonly four types of kidney stones have been identified namely: calcium oxalate, uric acid, struvite, and cystine ². Predominantly occurring are Calcium Oxalate Stones, resulting from the crystallisation of calcium ions and oxalate molecules within the urinary tract. Calcium containing stones may exist in the form of pure calcium oxalate (50%) or calcium phosphate (5%) or a mixture of both (45%) ^{1, 2, 3, 9}. Urolithiasis is a global health concern predicted to affect 1 in 10 people of a population by 2045 it primarily affects populations in the age of 20 to 45 years, affecting 11% men and 8% women worldwide ².

Urinary stones are the result of different mechanisms (low urine pH, hyperuricosuria, hyperoxaluria, high urinary calcium, low urine volume ^{3, 4}. Exceeding the supersaturation limit of the stone precursor in urine is the most widely accepted mechanism ³. Kidney stones formation occurs through four steps: crystal nucleation, crystal growth, crystal aggregation and crystal retention ⁵. Any crystal retained in the nephron can become a stone because it is a nidus of further crystal growth and deposition of other ionic species. The retention and growth of these crystals leads to the formation of insoluble stones. Hydration is the first step in medical management by increasing fluid intake sufficiently to produce 2–3 L/day of urine. Corticosteroids, phosphor-diesterase type 5 (PDE-5) inhibitors, calcium-channel blockers, and α-adrenergic blockers have been evaluated as part of the medical expulsive therapy ^{6, 7, 8}. Thiazide and other thiazide type diuretics have shown promising results in the treatment of urolithiasis. Citrate can inhibit kidney stone formation through a variety of mechanisms. It reduces the urinary supersaturation of calcium salts by forming soluble complexes with calcium ions and by inhibiting the crystal growth and aggregation. Extracorporeal shockwave lithotripsy (ESWL) is indicated preferentially in renal stones up to 2 cm in diameter ^{8, 9, 10}. Despite the availability of conventional treatments, urolithiasis remains a recurrent condition due to the lack of agents with true lithotriptic activity. Existing therapies primarily offer symptomatic relief without effectively dissolving existing stones. This highlights the need for safer, more effective alternatives. The present study aims to develop and evaluate a polyherbal oral medicated jelly combining traditionally used anti-urolithiatic herbs to explore its potential in dissolving calcium oxalate stones and offer a novel approach for the management of urolithiasis.

TABLE 1: TRADITIONAL NAMES AND CLASSIFICATION OF HERBAL DRUGS USED

TABLE 2: TRADITIONAL USE AND PHYTOCHEMICALS PRESENT IN HERBS USED

TABLE 3: STRUCTURES OF THE MAJOR PHYTOCONSTITUENTS USED

Bryotoxin	Bufadienolides	Tribuloside
Kaempferol	Rutin	Quercetin

MATERIALS: Cystone tablets were procured from Himalaya Drug Company (India) and served as the reference formulation to evaluate anti-urolithiatic activity. calcium oxalate (analytical grade) was used as a lithogenic agent. Phosphate buffer (IP grade) was prepared as per standard protocols and used to maintain the required pH during in vitro studies. Hydrochloric acid (HCl) and ammonia solution (NH₄OH) were used for decalcification of the egg membrane. Carrageenan was utilized as a gelling agent in the simulated formulation matrix. Sugar and sodium benzoate were incorporated as excipients in the preparation of syrup-based formulations. Agress 1100 HPLC system were used for HPLC analysis. CT3 Texture Analyzer (Brookfield Laboratories, Middleboro, USA) was used for texture analysis. 0.1Mv Lcd lab India pico digital pH meter was employed for pH measurement.

Methodology:

Extraction: Powdered Tribulus terrestris and crushed leaves of kalanchoe pinnata and cynodon dactylon were separately stored in airtight containers. The hydroalcoholic extract of each plant material was prepared by macerating the powdered plant parts in distilled water and 95% ethanol, and maintained in an incubator at 37°C for 24 h. Then, the samples were sonicated using an ultrasound sonicator (25-42 kHz waves) for one hour followed by maceration for 24 h. The herbal extracts were then filtered through Whatman filter paper using vacuum suction. The filtrate was then concentrated in an electric water bath to obtain the herbal extracts.

Formulation Development: Medicated jellies were prepared by using Carrageenan as a gelling agent in different concentrations as given in the formula Table 4. The gelling agents were tried initially at various concentrations to achieve desired appearance, stiffness and release. Citric acid is used to maintain the pH. Sodium benzoate is used as a preservative and sugar is used as sweetening and bulking agents.

The jellies are prepared by taking carrageenan, citric acid in beaked and heated with continuous stirring to get solution form. In another beaker sugar solution is prepared and transferred into the first beaker. Sweetening and flavouring agents were added and mixed thoroughly. The solution was transferred into moulds to avoid exposure to the outer environment. The formed jellies were stored in a dry place ²⁰.

TABLE 4: FORMULATION TRIALS OF VARYING CONCENTRATION OF GELLING AGENTS

Evaluation of Formulation:

Dissolution Test: The dissolution test was carried out using the paddle method with a stirring rate of 50 rpm at 37 °C. In 900 mL of Phosphate buffer pH 6.8 in dissolution apparatus (Electrolab). Phosphate buffer, pH 6.8, was prepared by mixing 250 mL of 0.2 M potassium dihydrogen phosphate solution with 118 mL of 0.2 M sodium hydroxide solution ²¹. After 5, 10, 15, 20, 40, 60, and 90 min of stirring, an aliquot of 5 mL of the sample was pipetted out and filtered through a membrane filter of pore size 0.2 µm then replaced with the same amount of fresh dissolution media ^{22, 23}.

The concentration of extracts was determined by UV spectrophotometer at λmax 268 nm and 238 nm.

Texture Analysis: Texture Profile Analysis (TPA) was carried out utilising a CT3 Texture Analyzer (Brookfield Laboratories, Middleboro, USA). This instrumental approach served as a robust method for evaluating the mechanical attributes of the prepared OMJ. Widely recognized as the "two-bite test," this procedure entails the controlled penetration of the probe into the sample twice, each time reaching a specific depth, before reverting to its original position. This systematic approach allowed for a comprehensive examination of the sample's textural properties, providing valuable insights into its structural integrity and sensory attributes. Different texture parameters like hardness, cohesiveness, springiness, gumminess, chewiness were measured from the obtained force–time plots. The TA5 kind of texture analysis probe has a constant speed of 1.0 mm/s and a trigger load of 1 g. The target was 4 mm, and the test type was compression test. There were two cycles in all. The test took place over a total of 12 minutes and 27 seconds. There were 50.00 points per second of data.

In-vitro Assessment:

Analysis of Lithotriptic Activity ²⁴: 

Isolation of Egg Membrane: The semipermeable membrane of eggs, nestled between the calcified outer shell and the inner contents of albumin and yolk, played a central role in our experimental approach. The semipermeable membrane of egg was obtained by chemical decalcification process by immersing eggs in 2M HCl overnight. This treatment ensured complete removal of the calcified shell, leaving the semi-permeable membrane intact. Following decalcification, the eggs were thoroughly washed with distilled water to eliminate any residual traces of the Acid. With precision, a small aperture was made at the top of the egg using a sharp pointer, facilitating the complete expulsion of the inner contents from the decalcified egg. Subsequently, the egg membrane was rinsed with distilled water, ensuring its pristine condition.

To optimise the membrane's properties, it was then immersed in an ammonia solution for a 20 minutes period, maintaining a moist environment and to neutralise residual acid. Following this treatment, the membrane was rinsed once again with distilled water to remove any excess ammonia traces. Finally, the prepared egg membranes were stored in the refrigerator at 4-5 degree in phosphate buffer at a pH range of 6.8 to 7.00 to preserve their structural and functional integrity for subsequent experimentation.

Incubation of Extracts with Experimental Kidney Stones (Calcium Oxalate Crystal): 10 mg of calcium oxalate with 100 mg of each of three extracts in combination was delicately encased within semi-permeable membranes ensuring controlled interaction within the experimental milieu and  suspended within a conical flask containing 100 ml of 0.1 M TRIS HCl buffer (pH=7). The samples were incubated at 37°C for a duration of 48 hours.100mg of standard Cystone with 10 mg of calcium oxalate was also treated in a similar manner as that of test extracts.

The experimental design encompassed three distinct groups. The first served as a negative control, consisting solely of 10 mg of calcium oxalate. The second group combined the calcium oxalate 10 mg with 100 mg of the standard compound (Cystone), while the third group employed calcium oxalate 10 mg.

HPLC Analysis: The literature lacks well-defined in-vitro techniques for accurately assessing lithotriptic effects. In this study, we aimed to develop a novel methodology for the estimation of calcium oxalate (CaOx) and to evaluate the lithotriptic efficacy of the test drug. Specifically, we established an innovative and previously unreported approach for CaOx quantification using high-performance liquid chromatography (HPLC) analysis.

In this study, we utilised HPLC to determine the level of calcium oxalate in the samples under investigation. The objective of the experiment was to quantitatively estimate the concentration of calcium oxalate in the provided samples using the Agres 1100 HPLC system. Two samples and one standard were prepared for analysis. A working standard solution ranging from 0 to 1000 micrograms per millilitre (µg/mL) was prepared in the base solution and filtered through a 0.22 micrometre syringe filter to ensure purity and uniformity.

The experiment began with the careful weighing of 10 mg of calcium oxalate, 100mg of each test drug and standard compound, Cystone. These components were delicately encased within semi-permeable membranes, ensuring controlled interaction within the experimental milieu. These were carefully packed together in a semi-permeable membrane, ensuring uniform distribution and minimal interference. Suspended within a conical flask containing 100 ml of 0.1 M TRIS buffer, the samples were subjected to controlled conditions, with an incubator set at 37°C for a duration of 48 hours. After incubation for two days, 1 millilitre (mL) of the sample solution was collected from the buffer, assuming that calcium oxalate crystals would diffuse out from the semi-permeable membrane due to the influence of the drug. The collected sample was then filtered through a 0.22 micrometre filter to remove any particulate matter or impurities before injection into the HPLC system.

The HPLC analysis was performed using an Agress 1100 HPLC system equipped with a UV-Visible detector. The injection volume for each sample was 20 microliters (µL), and the run time was set to 15 minutes. Isocratic gradient conditions were employed with a mobile phase consisting of methanol: water (50% methanol + 50% water + 0.1% formic acid). The flow rate was maintained at 1.0 millilitre per minute (mL/min) to ensure optimal separation and elution of the analytes. The HPLC analysis revealed distinct peaks corresponding to the retention time of calcium oxalate in both the samples and the standard solution.

Microbial Assay: Individuals suffering from urolithiasis are more susceptible to urinary tract infections (UTIs). Consequently, evaluating the antimicrobial activities of various extracts is crucial in addressing this increased risk. This study focuses on assessing the efficacy of these extracts in combating UTI-causing pathogens in urolithiasis patients.

Anti-bacterial and anti-fungal activity of the phytochemical extracts has been tested using the Agar well-diffusion method. Nutrient broth/agar was used to cultivate bacteria ²⁵. Escherichia coli, Staphylococcus aureus were the bacteria used and Candida albicans was the fungi used. These microbes are the causative agents in most of the cases of urinary tract infections (UTI). A 20 ml of molten nutrient agar media has been added into sterilised Petri-plates ²⁶. Agar plates were prepared and 0.1 ml volume of each diluted microbial isolates was transferred into the agar plates. The inoculated agar plates were kept for 30 min for the isolate to diffuse appropriately into the medium ²⁷. A 5 mm sterile cork borer was used to bore wells on the medium 0.1 ml volume of the K. pinnata extract and Wells are loaded with 100 mg/ml conc. of extract suspended in distilled water. The agar plates were labelled accordingly ²⁷. For 24 hrs the plates have been incubated at a temperature of 37C ²⁶. The Cystone tablet from Himalaya Drug Company was employed as a positive control as well as wells loaded with sterile water were used as a negative control. The inoculated agar plates were incubated properly and viewed for the diameter of inhibition zones. The diameter of inhibition zones was suggested by clear areas without growth around the well.

RESULT AND DISCUSSIONS:

HPLC Analysis:

Standards Analysis of HPLC: At a concentration of 10 micrograms per millilitre (µg/mL), no discernible peak was observed, indicating a sub-threshold concentration insufficient for detection under the specified chromatographic conditions. A standard solution of 100 µg/mL manifests a distinct peak at a retention time of 2.43667 minutes, with a corresponding peak area of 796.34. This observation corroborates the presence of the target compound at an intermediate concentration, indicative of its chromatographic detectability within the specified concentration range. Upon elevating the concentration to 1000 µg/mL, a prominent peak emerged at a retention time of 2.32833 minutes, exhibiting a substantially amplified peak area of 14215.58. This heightened peak intensity underscores the increased abundance of the target compound within the analytical matrix, affirming its robust detection at higher concentrations ²⁸.

Sample Analysis of HPLC: The standard marketed preparation, boasting a concentration of 1000 µg/mL, yielded a discernible peak at a retention time of 2.44083 minutes, accompanied by a peak area of 1130.82. This chromatographic signature aligns with the expected behaviour of a standard solution, reaffirming the fidelity of the analytical method employed. In contrast, the test drug sample, also prepared at a concentration of 1000 µg/mL, exhibited a distinct peak at a retention time of 2.28417 minutes, characterised by a peak area of 1448.66.

Notably, the observed retention time deviates slightly from the standard marketed preparation, indicative of potential variations in compound composition or formulation ²⁸.

FIG. 1: REFERENCE FORMULATION ACTIVITY OF CYSTONE (STANDARD: BLUE CAP)

FIG. 2: SAMPLE NAME – TEST GROUP ACTIVITY OF POLYHERBAL OMJ (TEST: RED CAP)

FIG. 3: STANDARD CALCIUM OXALATE 1000 µG/ML

FIG. 4: STANDARD CALCIUM OXALATE 100 µG/ML

Dissolution Test: The dissolution test serves as a pivotal component in assessing the release kinetics of pharmaceutical formulations, shedding light on the propensity of a drug to liberate from its dosage form over time.

In our investigation, we conducted a dissolution test for the formulated drug, monitoring its absorbance at wavelengths of 268 nm and 238 nm, with the respective readings depicted below:

TABLE 5: DISSOLUTION TIME AND OBSERVED ABSORBANCE OF THE ORALLY MEDICATED JELLIES

The dissolution profile, as delineated by the absorbance readings at 268 nm and 238 nm, portrays a discernible trend over the experimental time frame ²⁹. Notably, there is a progressive augmentation in absorbance values at both wavelengths with increasing time intervals, indicative of a concurrent increase in drug concentration within the dissolution medium.

This progressive escalation in absorbance, culminating in maximal readings at the 60 and 90-minute junctures, underscores the gradual and time-dependent release kinetics inherent to the formulation under scrutiny ^{29, 30}. The convergence of absorbance trends across both wavelengths underscores the robustness and consistency of our experimental observations, fortifying the veracity of our analytical approach. In summation, the dissolution test elucidates a concerted and time-dependent release of the drug from its formulation, thereby furnishing invaluable insights into its dissolution kinetics and therapeutic efficacy. These findings hold significant implications for pharmaceutical formulation design and optimization, facilitating informed decision-making in drug development endeavours.

Texture Analysis ^{31, 32}: In our investigation, we subjected the sample to comprehensive texture analysis, encompassing parameters such as hardness, cohesiveness, springiness, gumminess, chewiness, and pH.

Hardness: The sample exhibited a hardness value of 518 grams, indicative of its resistance to deformation or breakage under applied force. This parameter serves as a crucial determinant of textural quality, with higher hardness values often correlating with perceived firmness and structural integrity.

Cohesiveness: A cohesiveness value of 0.83 was observed, highlighting the sample's ability to maintain its internal structure upon deformation. Cohesiveness reflects the degree of internal bonding within the sample, with higher values suggestive of enhanced structural resilience and mouthfeel.

Springiness: The sample displayed a springiness value of 36.70, denoting its propensity to regain its original shape following deformation.

Springiness underscores the elasticity of the sample, offering insights into its resilience and bounce-back characteristics upon mechanical manipulation.

Gumminess: With a gumminess value of 431.00 grams, the sample exemplified its chewy and adhesive nature. Gumminess quantifies the energy required to disintegrate the sample during mastication, with higher values indicative of increased chewiness and oral adhesiveness.

Chewiness: The sample exhibited a chewiness value of 155.00 millijoules (mJ), underscoring its textural complexity and resistance to breakdown during mastication. Chewiness amalgamates the concepts of hardness and gumminess, encapsulating the overall effort required to masticate the sample into a swallow able consistency.

pH: The pH value of the sample was measured at 4.5, serving as a crucial determinant of its acidity or alkalinity. pH exerts a profound influence on the sensory attributes and stability of food products, with deviations from optimal pH ranges potentially impacting flavour, texture, and shelf-life.

FIG. 5: TEXTURE ANALYSIS GRAPH

Cohesiveness and springiness, reflective of the sample's internal integrity and elasticity, contribute to its mouth feel and sensory appeal, ensuring a pleasurable eating experience characterised by tactile satisfaction and resilience. The pronounced gumminess and chewiness underscore the sample's adhesive and chewy nature, endowing it with a distinctive textural complexity that may evoke sensory intrigue and prolong oral processing. Furthermore, the measured pH value assumes significance in elucidating the sample's acidity level, with potential ramifications for flavour perception, microbial stability, and shelf-life.

Anti Microbial Assay: The antimicrobial activity of Kalanchoe pinnata (K. pinnata) extract was evaluated using the agar well-diffusion method. The extract's efficacy was tested against Escherichia coli, Staphylococcus aureus, and Candida albicans, which are common causative agents of urinary tract infections (UTIs). The zones of inhibition, representing the antimicrobial effectiveness of the K. pinnata extract, were measured and compared to those of a positive control (Cystone tablet) and a negative control (sterile water).

The largest zone of inhibition (2.08 cm) was observed against E. coli, indicating strong antibacterial activity. E. coli is a common pathogen in UTIs, and the significant inhibition zone suggests that K. pinnata extract could be a potent alternative or adjunctive treatment for E. coli-induced UTIs. The zone of inhibition against S. aureus was 1.0 cm. Although smaller than that observed for E. coli, this inhibition is still notable. S. aureus is a less frequent but possible cause of UTIs. The moderate activity against this bacterium suggests that while K. pinnata extract has some efficacy, it may be more potent against gram-negative bacteria like E. coli. The extract showed a zone of inhibition of 1.9 cm against C. albicans, indicating substantial antifungal activity. C. albicans is a common fungal pathogen in UTIs, particularly in patients with complicated infections or those who are immunocompromised. The results suggest that K. pinnata extract could be beneficial in managing fungal UTIs ^{32, 33}.

FIG. 6: ZONE OF INHIBITION OF CANDIDA ALBICANS

FIG. 7: ZONE OF INHIBITION OF E. COLI                     

FIG. 8: ZONE OF INHIBITION OF STAPHYLOCOCCUS AUREUS

The use of Cystone tablets as a positive control validates the methodology, as Cystone is known for its antimicrobial properties. The absence of zones of inhibition around the wells containing sterile water confirms that the observed antimicrobial activity is due to the K. pinnata extract.

CONCLUSION: The comprehensive investigation conducted in this study provides significant insights into the chromatographic, dissolution, textural, and antimicrobial characteristics of the tested compounds and formulations. HPLC analysis demonstrated the detectability of the target compound across a broad concentration range, establishing the method's sensitivity and linearity. The dissolution profile indicated a time-dependent release of the drug, confirming the formulation's efficacy for sustained therapeutic delivery. Texture analysis revealed significant hardness, cohesiveness, springiness, gumminess, and chewiness, contributing to the sample's overall quality, with a pH value of 4.5 supporting microbial stability and sensory attributes. The Kalanchoe pinnata (K. pinnata) extract exhibited notable antimicrobial activity, particularly against E. coli, highlighting its potential as an alternative or adjunctive treatment for UTIs. These findings have critical implications for pharmaceutical formulation and drug development, emphasising the need for a multifaceted approach to ensure efficacy, quality, and therapeutic potential. Further research is recommended to optimize formulations and explore additional therapeutic applications.

ACKNOWLEDGEMENTS: Sincere gratitude to Lifesenz cancer research Labs Pvt Ltd. for providing the facility for HPLC analysis. We are thankful to Gautami Sabat, Sunil Patil Rutuja Belwalkar and Om Dhurifor their unwavering support throughout study and research writing. The research did not receive any external funding from any agencies in public, commercial or non-profit sectors.

CONFLICT OF INTEREST: There is no conflict of interest for this manuscript. The authors declare that they have no known competing financial interest, directly or indirectly.

REFERENCES:

1. Thakore P and Liang TH: Urolithiasis. [Updated 2023 Jun 5]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK559101/
2. https://www.kidney.org/atoz/kidneystones.
3. Allam EA: Urolithiasis unveiled: pathophysiology, stone dynamics, types, and inhibitory mechanisms: a review. African Journal of Urology 2024; 30(1): 34.
4. Sakhaee K: Recent advances in the pathophysiology of nephrolithiasis. Kidney International 2009; 75(6): 585-595.
5. Shtukenberg AG, Hu L, Sahota A, Kahr B & Ward MD: Disrupting crystal growth through molecular recognition: designer therapies for kidney stone prevention. Accounts of Chemical Research 2022; 55(4): 516-525.
6. Jahrreiss V, Seitz C & Quhal F: Medical management of urolithiasis: Great efforts and limited progress. Asian Journal of Urology 2024; 11(2): 149-155.
7. Patil S, Sanman KN, Prabhu L, Shetty R, Dani T & Jebaraj A: Comparison of tamsulosin, naftopidil and tadalafil as medical expulsive therapy for lower ureteric calculus: An observational study 2021.
8. Demoulin N, Aydin S, Gillion V, Morelle J & Jadoul M: Pathophysiology and management of hyperoxaluria and oxalate nephropathy: a review. American Journal of Kidney Diseases 2022; 79(5): 717-727.
9. Amin R, Kapoor P & Silva A: Pharmacologic treatment of kidney stones: strategies for prevention and tailored therapies. Journal of Nephrology & Therapeutics 2023; 13(4): 115–125.
10. Allam EAH: Urolithiasis unveiled: pathophysiology, stone dynamics, types, and inhibitory mechanisms: a review. African Journal of Urology 2024; 30: 34.
11. Dogra P, Sharma K, Bharti J, Kumar N & Kumar D: Kalanchoe pinnata is a miraculous plant: a review. J Biomed Allied Res 2023; 10.
12. Singh RK, Garg A & Shrimali K: Botanical description, photochemistry, traditional uses, and pharmacology of the “wonder plant”. Kalanchoe pinnata (Linn.) Pers: an updated review. Journal of Pharmaceutical and Medicinal Chemistry 2022; 8(1).
13. Mejía-Méndez JL, Sánchez-Ante G, Minutti-Calva Y, Schürenkämper-Carrillo K, Navarro-López DE, Buendía-Corona RE, González-Chávez MDCÁ, Sánchez-López AL, Lozada-Ramírez JD, Sánchez-Arreola E & López-Mena ER: Kalanchoe sp. extracts—Phytochemistry, cytotoxic, and antimicrobial activities. Pharmaceuticals (Basel) 2024; 17(8): 1051.
14. Chhatre S, Nesari T, Somani G, Kanchan D & Sathaye S: Phytopharmacological overview of Tribulus terrestris. Pharmacognosy Reviews 2014; 8(15): 45–51.https://doi.org/10.4103/0973-7847.125530
15. Ștefănescu R, Tero-Vescan A, Negroiu A, Aurică E & Vari CE: A comprehensive review of the phytochemical, pharmacological, and toxicological properties of Tribulus terrestris Biomolecules 2020; 10(5): 752.
16. Shendye NV & Gurav SS: Cynodon dactylon: A systemic review of pharmacognosy, phytochemistry and pharmacology. International Journal of Pharmacy and Pharmaceutical Sciences 2014; 6(8): 7-12.
17. Das S, Morya S, Neumann A & Chattu VK: A Review of the Pharmacological and Nutraceutical Properties of Cynodon dactylon. Pharmacognosy Research 2021; 13(3).
18. Al-Snafi AE: Chemical constituents and pharmacological effects of Cynodon dactylon-A review. IOSR Journal of Pharmacy 2016; 6(7): 17-31.
19. Parihar S & Sharma D: Cynodon dactylon: A Review of Pharmacological Activities. Sch Acad J Pharm 2021; 11: 183-189.
20. Prasanthi N, Nandini P, Manikiran S & Ramarao N: Design and Development of Herbal Pediatric Edible Jelly for Anthelmintic Infections (11th Ed). Journal of Pharmaceutical Sciences and Research 2019; 2417-2421.
21. Uchiyama H, Nogami S & Katayama K: Jelly containing composite based on α-glucosyl stevia and polyvinylpyrrolidone: Improved dissolution property of curcumin. European Journal of Pharmaceutical Sciences 2018; (117): 48-54. https://doi.org/10.1016/j.ejps.2018.02.011
22. Kim KH, Jun M & Lee MK: Bioavailability of the Common Cold Medicines in Jellies for Oral Administration. Pharmaceutics 2020; 12: 1073. https://doi.org/10.3390/pharmaceutics12111073
23. Prasanthi N, Nandini P, Manikiran S & Ramarao N: Design and Development of Herbal Pediatric Edible Jelly for Anthelmintic Infections (11th Ed. Journal of Pharmaceutical Sciences and Research 2019; 2417-2421.
24. Bansode P, Pawar P & Babar M: In-vitro Urolithiatic activity of Bryophyllum pinnatum against experimentally designed calcium oxalate and calcium phosphate stones. British Journal of Pharmaceutical and Medical Research 2016; 1(1): 34-40. http://www.bjpmr.org
25. Pattewar SV & Patil DN: Formulation of herbal antibacterial cream by using extract from Kalanchoe pinnata leaves. Research Journal of Topical and Cosmetic Science 2014; 5(1): 1-4.
26. Sitrarasi R, Ravindran B, Uma Maheshwari Nallal V, Razia M, Jin Chung W, Shim J, Chandrasekaran M, Dwiningsi Y, Rabab AR, Alkahtani J, Elshikh MS & Ovi D: Inhibition of multi-drug resistant microbial pathogens using an ecofriendly root extract of Furcraea foetida mediated silver nanoparticles. Journal of King Saud University – Science 2021; 34.
27. Ehwarieme DA, Offuah B & Ilondu EM: Evaluation of the phytochemical and antimicrobial activity of Kalanchoe pinnata against plant pathogens isolated from diseased plant pathogens. FUDMA Journal of Sciences 2021; 5(3): 310-314. https://doi.org/10.33003/fjs-2021-0503-755.
28. Shen Y, Luo X, Li H & Cheng L: Evaluation of a high-performance liquid chromatography method for urinary oxalate determination and investigation regarding the pediatric reference interval of spot urinary oxalate to creatinine ratio for screening of primary hyperoxaluria. Journal of Clinical Laboratory Analysis 2021; 35(6).
29. Smith A & Johnson R: Release kinetics and dissolution profiling of tablet formulations: method development and comparison at multiple wavelengths. International Journal of Pharmaceutics 2023; 615.
30. Lee H & Patel A: Evaluating dissolution dynamics of immediate release tablets using UV spectrophotometry: implications for formulation design. European Journal of Pharmaceutical Sciences 2022; 169: 106070.
31. Li X, Wong T & Miller B: Advances in characterization of transdermal and topical products: a systematic review of texture analysis methods. AAPS Pharm Sci Tech 2025.
32. Kumar S & Garcia M: A versatile tool for pharmaceutical evaluation of solid oral dosage: texture profile analysis and its correlation with formulation quality. International Journal of Pharmaceutics 2023; 638: 122632.
33. Kumar G, Kumar R & Rana H: The pharmacological activities of mother of thousand (Kalanchoe pinnata). Research Journal of Pharmacology and Pharmacodynamics 2023; 15(1): 31–35.
34. Anandan J & Shanmugam R: Antioxidant, anti inflammatory, and antimicrobial activity of the Kalanchoe pinnata and Piper longum formulation against oral pathogens. Cureus 2024; 16(4).
    

    ---

    How to cite this article:

    Mule SS, Naik HA, Solanki KR, Devnani L and Sanaye M: Formulation and evaluation of a novel polyherbal oral medicated jelly for treatment of urolithiasis. Int J Pharm Sci & Res 2025; 16(12): 3324-34. doi: 10.13040/IJPSR.0975-8232.16(12).3324-34.

    ---

    All © 2025 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.