Formulation and Evaluation of Kalanchoe Pinnata Syrup: A Pharmaceutical Dosage Form for Treatment Urolithiasis (Renal calculus) and UTI

Herbal medicines have gained significant attention in recent years due to their natural origin, minimal side effects, and broad therapeutic potential. Among these, Kalanchoe pinnata (Bryophyllum pinnatum) holds a prominent place in traditional medicine for its diverse pharmacological properties, including anti-inflammatory, antimicrobial, nephroprotective, and wound-healing activities. This plant is rich in bioactive compounds such as flavonoids (quercetin, kaempferol), alkaloids, phenolic acids (caffeic acid, ferulic acid), triterpenoids, and bufadienolides, which contribute to its therapeutic potential.   Renal calculus (kidney stones) and UTI (urinary tract infection) inflammation of the urinary tract are prevalent health issues that significantly impact quality of life. Existing treatments, while effective, are often associated with high costs and adverse effects, driving the demand for safer, more affordable alternatives. Kalanchoe pinnata has demonstrated significant potential in dissolving renal calculi, reducing inflammation, and providing nephroprotection due to its bioactive components.   Traditional preparations of Kalanchoe pinnata, such as teas or infusions, face challenges in terms of stability, standardization, and patient compliance. Therefore, this study aims to develop a stable and patient-friendly Kalanchoe pinnata syrup for the management of UTI and Urolithiasis (Renal calculus). By utilizing excipients compliant with the Indian Pharmacopoeia (IP), this formulation bridges the gap between traditional remedies and modern pharmaceutical practices, offering an innovative and effective dosage form for urinary and renal health management.

MATERIALS & METHODS

Plant collection

Kalanchoe Pinnata Plant leaf were collected in Dusanapudi village garden in the month of November to December.

The plant was authenticated by the botanist of Dr. C.S.N Degree and Pg College, Bhimavaram, Andhra Pradesh, India.

Ethanolic Extract of Kalanchoe pinnata

Extraction Process [1]

• Kalanchoe pinnata leaves were cleaned, cut, and oven-dried at 50°C for 3 days.
• The dried leaves were pulverized into a fine powder.
• 10 g of the dry powder was mixed with 100 mL ethanol (96%) and stirred for 24 hours.
• The extract was then filtered and concentrated.

Phytochemical Constituents of Ethanolic Extract (EE)

• Saponins & flavonoids ? Prevent calcium oxalate crystallization (anti-urolithiatic).
• Alkaloids & tannins ? Antimicrobial activity against UTI pathogens.
• Steroids & phenolic compounds ? Reduce renal inflammation.

Table 2: Antioxidant Activity (DPPH, NO, Superoxide Assays) [1]

• K. pinnata ethanolic extract moderately inhibits free radicals, supporting nephroprotective effects in urolithiasis.
• The DPPH radical scavenging (49.5%) suggests it helps reduce oxidative stress in kidney cells.

4. Anti-Inflammatory Activity

•The ethanolic extract (EE) of K. pinnata showed mild anti-inflammatory effects compared to standard diclofenac.

•Steroidal compounds may contribute to reduced inflammation in UTI and kidney disorders.

Anti-urolithiatic properties ? Saponins & flavonoids prevent kidney stone formation.
Anti-UTI effects ? Alkaloids & tannins inhibit bacterial growth.

Nephroprotective & diuretic action ? Supports kidney function.

The extract was then filtered and concentrated

Table 3: Ingredients and Formulation of Kalanchoe pinnata Leaf Syrup (F1)

This  Formulation is failed due to ph it is more acidic

This formulation is slightly acidic and stability issues (F2) is good compared to (F1)

This Formulation (F3 ) has neutral ph and stability this is perfect compared to F1 and F2

The optimized Kalanchoe pinnata syrup formulation with 10 mL extract, trisodium citrate (1 g/10 mL), and glycerin (2.5 mL) showed neutral pH and superior stability compared to previous formulations. Unlike the first formulation (20 mL extract, citric acid), which had an acidic pH of 3.84, and the second formulation, which had a pH of 5.55, the final formulation maintained better organoleptic properties and antimicrobial activity. This optimized syrup is more effective, stable, and patient-compliant for urolithiasis and UTI prevention.

1. Preparation of Simple Syrup:
   • Take a clean, heat-resistant beaker or container.
   • Measure 66.7 gm of sucrose and place it into the container.
   • Add 33.3 ml of distilled water to the sucrose.
   • Heat the mixture gently using a burner until the sucrose is completely dissolved in the water. Stir occasionally to ensure uniform dissolution.
   • Once all the sucrose has dissolved and the solution is clear, remove the container from the heat source.
   • Allow the solution to cool to room temperature.
2. Formulation of the Syrup:
   • Once the simple syrup is cooled, measure out 75 ml of the prepared simple syrup.
   • In a clean container, combine the following:
     • 10 ml of Kalanchoe pinnata leaf ethanolic extract (containing the active compounds).
     • 10 ml of tri sodium citrate solution (for pH adjustment and flavor).
     • 2.5 ml of glycerine (Humectant, viscosity enhancer, improves taste)
     • 2.5 ml of sodium benzoate solution (as a preservative).
3. Mixing:
   • Add the 75 ml of simple syrup to the container with the ethanolic extract, tri sodium citrate, glycerine and sodium benzoate.
4. Final Volume and Storage:
   • After mixing, ensure the total volume is 100 ml.
   • Transfer the prepared syrup into an amber-colored glass bottle to protect it from light.
   • The formulation showed a decrease in calculi size, indicating potential stone breakdown.

2.Anti-Microbial Activity (Against E. coli)

oExhibits bactericidal effects against Escherichia coli, a common UTI-causing pathogen.

oThe optimized syrup formulation led to a significant reduction in E. coli count, proving its efficacy in preventing UTIs.

oThe presence of sodium benzoate enhances microbial stability and extends shelf life.

Kalanchoe pinnata extract and tri sodium citrate helps in Urolithiasis, Cystitis, UTI

Evaluation of Kalanchoe Pinnata Syrup Formulation (F3)

Table 6: organoleptic properties of Kalanchoe pinnata leaf syrup formulation (F3):

Physical evolution of kalanchoe pinata leaf syrup formulation (F3)

1. Density:

• Procedure:
  1. Take a clean, dry, and calibrated pycnometer or a suitable density bottle.
  2. Weigh the empty pycnometer (W1).
  3. Fill the pycnometer with a known volume of the syrup formulation.
  4. Weigh the filled pycnometer (W2).
  5. Calculate the density using the formula:

Density=Weight of the syrup (W2 - W1)?/Volume of the syrup

2. Viscosity:

Ostwald Viscometer:

1. Preparation:
   • Ensure the Ostwald viscometer is clean and dry before use.
   • Set the viscometer vertically using a stand to ensure it is stable.
   • Check that the capillary tube is not obstructed or damaged.
2. Filling the Viscometer:
   • Fill the Ostwald viscometer with the syrup sample. Make sure the liquid level is above the upper bulb but below the marked upper graduation.
   • Avoid any air bubbles in the capillary tube.
3. Timing the Flow:

• Using a stopwatch or timer, allow the syrup to flow between the two marked points (upper and lower marks) in the capillary tube. Start the timer as the meniscus passes the upper mark, and stop it once the meniscus reaches the lower mark.
• Repeat the measurement two to three times to obtain an average time.

4. Calculation of Viscosity:

The viscosity can be calculated using the formula:

?=(t_s/t_std) ×?_std

Where_:

• ? = viscosity of the sample (in cP)
• t_s= time taken for the syrup to flow between the two marks
• t_std= time taken for a standard liquid (usually water) to flow between the same marks under identical conditions
• ?_std= viscosity of the standard liquid (in cP)

Anti Microbial tests of the ethanolic extract kalanchoe pinnata syrup [4]

• Broad-Spectrum Activity:
  Ethanolic extracts of Kalanchoe pinnata leaf syrup show inhibitory effects against both Gram-positive and Gram-negative bacteria, as well as antifungal activity against yeast and mold species.
• Active Constituents:
  The antimicrobial effects are likely due to bioactive compounds such as flavonoids present in the ethanolic extract, which are key contributors to the observed activity.