INTRODUCTION:

Urolithiasis is a condition characterized by the formation of stones within the urinary tract, which may occur in the kidneys, ureters, bladder, or urethra. It is a multifactorial metabolic disorder with diverse etiologies, commonly leading to the development of kidney stones. Despite extensive research, the precise causes and mechanisms underlying stone formation in urology remain poorly understood.^1–3 In industrialized countries, the annual incidence of urolithiasis is estimated to be around 0.2%. Lifetime risk varies across regions, with figures ranging from 2–5% in Asia, 8–15% in Western countries, and reaching as high as 20% in Saudi Arabia. Recurrence rates are notable, with 10–23% of individuals experiencing recurrence within one year, 50% in five to years, and 75% in 20 years². Data from the Rochester Epidemiology Project revealed a 4% yearly rise in pediatric renal stone cases between 1984 and 2008, with a much significant increase observed in older children, specifically those aged 12 to 17 years. Additionally, a population-based study conducted in South Carolina from 1997 to 2012 found a 16% yearly increase in the incidence of kidney stones, especially in younger female patients between the ages of 10 and 24⁴.

Urolithiasis represents a substantial economic burden, with costs ranging from £190 million to £324 million in England for the year 2010. In the United States, it is projected that the annual financial impact will increase by an additional $1.24 billion by 2030⁵. A study by the Società Italiana di Medicina Generale (SIMG) in 2006 found that among urolithiasis patients, 19% sought urological consultation, 4.6% required hospitalization, 48.8% underwent ultrasonography (US), 7.2% had urography, 2.6% received non-contrast enhanced computed tomography (NCCT), and 3.4% underwent kidney-ureter-bladder radiography (KUB)⁶. Certain medications, including sulphonamides, ceftriaxone, triamterene, felbamate, and furosemide, have been associated with an increased incidence of urolithiasis⁴. In the initial evaluation of pediatric urolithiasis, ultrasonography (US) is recommended as the primary imaging modality, with non-contrast computed tomography (CT) serving as the secondary option. For renal stones greater than 20mm, shockwave lithotripsy (SWL) is considered the first-line treatment⁷. Mortality rates associated with interventional chronic diseases (ICD) have significantly decreased in last few decades, largely due to the widespread adoption of advanced treatment techniques, including extracorporeal shock wave lithotripsy (ESWL), contact lithotripsy (CLT), and percutaneous nephrolitholapaxy.

However, ESWL is still linked to potential kidney injury. In severe cases, this can result in the formation of intraparenchymal, subcapsular, or perirenal hematomas, which may lead to renal tissue sclerosis and impaired kidney function. Additionally, severe hydronephrosis and ureteral strictures are recognized as risk factors that may complicate the course of treatment following shockwave lithotripsy^3,8. A notable disadvantage of retrograde intrarenal surgery (RIRS) is the steep learning curve associated with mastering flexible ureteroscopy (f-URS), which may present challenges for some healthcare providers. While advancements in technology for percutaneous nephrolithotomy (PCNL) have made the procedure less invasive while maintaining a high stone-free rate (SFR), PCNL still carries a significant complication risk. As per the CROES in the PCNL Global Study, complication rates comprise postoperative fever (20.7%), renal pelvis perforation (6.7%), hemorrhage (16.1%), blood transfusion (11.9%), and procedure failure (3.5%)⁹. The risk factors for urolithiasis include age, gender, dietary habits, climate and seasonal variations, occupational influences, genetic predisposition, and the presence of comorbid conditions such as diabetes, obesity, hypertension, gout, and metabolic syndrome^10–14.

The inclusion of trace elements in the crystal lattice or within the layers of urinary stones may aid in their fragmentation. Trace elements like strontium, copper, nickel, iron, and zinc have the potential to bind with oxalate and phosphate ions form poorly soluble salts. Both copper and magnesium play a beneficial role in preventing urolithiasis. Magnesium acts as an inhibitor of kidney stone formation by binding to the crystal surface, thereby preventing the attachment of new ions and inhibiting their aggregation. Copper, even at very low concentrations, has been shown to influence the growth of calcium oxalate crystals^15,16. Supersaturation of stone-forming stuffs in the urine and blood leads to the development of microlites and salt crystals, creating an environment conducive to stone formation. The precipitation of these salts in the urine is restricted by numerous factors, such as cobalt ions, zinc, magnesium, hippuric acid, manganese, citrates, as well as the concentration of hydrogen ions, which normally ranges from 5.6 to 6.0 in urine.¹⁷

Medications used to treat hypercalciuric calcium stones in urolithiasis can induce number of side effects, including hypotension, hypokalemia, and hypocitraturia, in patients undergoing conventional drug therapy. This has sparked growing interest in the use of traditional herbal plants for the prevention and management of urolithiasis, owing to their relatively mild side effects. Herbal remedies offer potential benefits in preventing stone formation and alleviating associated inflammation and pain, largely due to their bioactive compounds^18–21. These compounds, such as flavonoids, alkaloids, saponins, sterols, and polyphenols, exhibit properties including anti-inflammatory, antioxidant, diuretic, and analgesic effects.²² Medicinal plants have been widely accepted since ancient times due to numerous advantages, including their low toxicity, safety, effectiveness, affordability, reduced likelihood of disease recurrence, and easy availability in rural areas.^23–26. Some plants with known anti-urolithiasis activity include Acalypha indica, Aerva lanata, Boerhaavia diffusa, Cynodon dactylon, Pedalium murex, Phyllanthus niruri, Solanum virginianum, Solanum xanthocarpum, and Terminalia arjuna.^27–40

The objective of this study is to evaluate the concentrations of copper and magnesium in ten selected anti-urolithiasis plants. This is based on the critical role that copper and magnesium concentrations play in the anti-urolithiasis properties of plants. Furthermore, the study aims to identify the most effective anti-urolithiasis plant by assessing the levels of these two minerals.

MATERIALS AND METHODS^41–43

Plant collection and the initial preparation:

The samples of the plant materials were freshly gathered from the forest range of sathuragiri hills and campus of Kalasalingam university. The plant samples parts were separated and thoroughly washed in pure water to remove the sand or dust particles and all other surface contamination. The plant materials were shade dried at ambient temperature. All the dried plant parts were mechanically pulverized and then the ground into fine powder. The powdered samples were kept at room temperature in closed dry plastic containers for the further elemental analysis.

Sample ashing procedure:

The Dry powder of the plant material was weighed and placed in a ceramic crucible and roasted in an electric muffle furnace at 450^oc for five hours. Ashing have wiped out all the organic components present in the sample. The crucible containing ash was then drawn from the muffle furnace and placed in the desiccator. Then the ashes were allowed to cool, and their weights were taken. The weights of the crude sample and ash are presented in Table 1.

Sample preparation for AAS:

Each sample (0.5 g) was weighed accurately and transferred into a 100-mL PTFE vessel. Subsequently, 5 mL 65% HNO3 was added to the sample. The resulting mixture was gently heated in a water bath at 90 °C. This process was continued for 1–2 h or until the solution became clear.

Whatman 42 was used to filter the samples (2.5-μm particle retention). Subsequently, deionized water was added in sufficient quantity to achieve a final volume of 100 ml. The resultant solutions were utilized for atomic absorption spectroscopy analysis.

Atomic absorption spectroscopical study:

The prepared samples of the dilute filtrate were utilized for elemental analysis via atomic absorption spectroscopy (AAS). An AA6300 model instrument was employed for the atomic absorption spectroscopy measurements. An appropriate hollow cathode lamp was utilized for the analysis. The concentrations of various elements were determined using the relative method, employing A.R. grade solutions of the elements of interest. The standard conditions for atomic absorption measurements are presented in Table 2.

The AAS apparatus was adjusted to the proper conditions for each element. The absorbance of each element was measured by aspirating the standard and sample solutions into flames. Using the software Wizard (Ver. 1.03), the absorbance for each sample solution and standard solutions was determined and noted. A calibration curve for each element was generated by plotting the absorbance versus the standard solution concentrations. The software programs provided the concentrations of the diluted samples directly. The concentration values for all the diluted samples were gathered by multiplying with their respective dilution factors.

Table No - 1: weights of the crude sample and ash

S. No	Plant name	Weight of dried powder taken	Weight of ash obtained
1	Pedalium murex	10 g	0.842g
2	Terminalia arjuna	10 g	0.526g
3	Aerva lanata	10 g	0.762g
4	Phyllanthus nirui	10 g	0.741g
5	Boerhavia diffusa	10 g	0.967g
6	Cynodon dactylon	10 g	0.685g
7	Solanum virginianum(new) solanum xanthocaroum(old)	10 g	0.585g
8	Acalypha indica	10 g	0.698g
9	Trachyspermum ammi	10 g	0.735g
10	Asparagus racemosus	10 g	0.812g

Table No - 2: Spectroscopical Parameters

S. No	Parameter	Magnesium	Copper
1	Wavelength	285.2	324.8
2	Lamp current low peak (mA)	8	6
3	Slit width(nm)	0.7	0.7
4	Lamp mode	NON-BGC	BGC-D2

RESULTS:

Upon analysis of the trace elemental composition of ten anti-urolithiatic plants, it reveals that magnesium and copper were present in the following quantities in all the ten plants at varying levels, as presented in Table 3 and figure 1. The results of this study indicate that the highest concentrations of magnesium were found in plant species Trachyspermum ammi, Pedalium murex, Aerva lanata, Boerhavia diffusa, and Asparagus racemosus, with concentrations of 0.8334, 0.8210, 0.7766, 0.7415, and 0.7258, respectively. Similarly, the highest concentrations of copper were observed in plant species Pedalium murex, Solanum virginianum, Asparagus racemosus, Cynodon dactylon, and Trachyspermum ammi, with corresponding concentrations of 2.1036, 1.4336, 1.3472, 1.0965, and 0.0923, respectively.

Table 3: concentration of Mg and Cu in ppm

S. No	Plant name	Conc of magnesium in ppm	Conc of copper in ppm
1	Pedalium murex	0.8210	2.1036
2	Terminalia arjuna	0.3435	0.0899
3	Aerva lanata	0.7766	0.6393
4	Phyllanthus nirui	0.3546	0.8385
5	Boerhavia diffusa	0.7415	0.3755
6	Cynodon dactylon	0.2905	1.0965
7	Solanum virginianum	0.6941	1.4336
8	Acalypha indica	0.5891	0.1195
9	Trachyspermum ammi	0.8334	0.0923
10	Asparagus racemosus	0.7258	1.3472

Figure 1: Concentration of copper and magnesium in ten selected plants

DISCUSSION:

Magnesium has been recognized as a potential inhibitor in the formation of kidney stones. In vitro experiments have indicated that higher-than-normal concentrations of magnesium can effectively reduce both the nucleation and development of calcium oxalate crystals, which are the primary components of kidney stones. Furthermore, supplementation with magnesium, especially when paired with vitamin B6, has been demonstrated to significantly decrease the likelihood of developing kidney stones. These findings underscore the protective role of magnesium in the crystallization process associated with urolithiasis⁴⁴. Supplementation with magnesium and vitamin B6 has been shown to significantly reduce the likelihood of developing kidney stones. Magnesium plays a key role in inhibiting the crystallization of calcium oxalate in human urine, a crucial step in the likelihood of developing kidney stones. Additionally, magnesium helps to limit the absorption of dietary oxalate from the gastrointestinal tract, further contributing to lover the level of oxalate in urine and reducing the potential for stone formation ⁴⁵. Magnesium acts as an inhibitor in the development of kidney stones by interacting with the crystal surface, preventing new ions from attaching and thus inhibiting further aggregation. Due to its smaller ionic radius, magnesium (Mg²⁺) binds to oxalate more readily than calcium (Ca²⁺), occupying the available binding sites and reducing the rate of oxalate crystallization. This interaction is concentration-dependent, as higher concentrations of Mg²⁺ enhance its ability to bind oxalate ions, limiting the capacity of calcium to compete for binding sites. As a result, the presence of magnesium can effectively reduce the formation and growth of calcium oxalate crystals¹⁶.

Copper (Cu) has been shown to hinder the growth of calcium phosphate crystals, although it does not appear to affect the crystallization of oxalate. In animal models, such as rachitic rats, copper exerts an inhibitory effect on the mineralization process. Additionally, studies have indicated that the concentration of copper stored in kidney stones is more significant than that of zinc, particularly in the case of oxalate stones. This suggests a potential role for copper in the development and composition of kidney stones, especially those containing oxalate^44,45. Copper (Cu) has been noted to have effect on the growth of calcium oxalate crystals, particularly at extremely low concentrations. As an essential antioxidant, copper is predominantly concentrated in the brain, liver, heart, and kidneys, where it plays a vital role in protecting cells from oxidative damage. Its presence in these organs at high concentrations highlights its physiological significance and potential involvement in processes related to crystal formation and kidney stone development ¹⁶. Stones that contained higher concentrations of trace elements, including copper, zinc, and magnesium, were found to be more fragile and exhibited increased susceptibility to disintegration during extracorporeal shock wave lithotripsy (ESWL). This suggests that the presence of these trace elements may influence the structural integrity of kidney stones, potentially enhancing their fragmentation during treatment⁴⁶. Trace elements like strontium, copper, nickel, iron, and zinc have the potential to bind with oxalate and phosphate ions form poorly soluble salts. These interactions can influence the crystallization process and contribute to the formation of mineral deposits, such as kidney stones 15⁴⁷.

Trachyspermum ammi, Pedalium murex, and Asparagus racemosus are plants known for their high concentrations of magnesium and copper, which contribute to the inhibition of urolithiasis. Previous studies indicate that Asparagus racemosus helps prevent lithiasis by increasing magnesium levels, thereby promoting the dissolution of stone formation. Additionally, the presence of bioactive compounds such sterols (4, 6-dihydryxy-2-O (-2- hydroxy isobutyl) benzaldehyde), Aspargamine A, flavanoids (quercetin, rutin and hyperoside), polysaccharides in these plants have been demonstrated to reduce oxidative stress. These compounds could have an impact in preventing the adhesion of crystals to renal endothelial cells, further inhibiting the development of kidney stones²².

Pedalium murex has shown potential in inhibiting the growth and aggregation of struvite crystals. It has been observed to reduce levels of potassium, creatinine, sodium, and uric acid, all of which are closely linked to kidney function and the formation of stones. This modulation of biochemical markers indicates that Pedalium murex may contribute to improved renal function and promote diuresis, which could further support the prevention of kidney stone formation ³². The anti-urolithiasis properties of Trachyspermum ammi may be attributed to several mechanisms, including its diuretic effects, inhibition of calcium oxalate (CaOx) crystal aggregation, antioxidant activity, protection of renal epithelial cells, and antispasmodic effects. These diverse actions suggest that Trachyspermum ammi holds therapeutic potential in the management of urolithiasis, a condition for which effective non-invasive treatment options are currently limited in modern medicine³⁷.

CONCLUSION:

In conclusion, the findings of this study highlight t the important role trace metals like copper and magnesium play in managing and preventing urolithiasis. It has been demonstrated that magnesium efficiently prevents the production of calcium oxalate crystals, mainly by restricting the absorption of oxalate from the gastrointestinal system and by decreasing crystal nucleation and growth. This activity presents a promising strategy to lower the risk of kidney stone development, especially when combined with vitamin B6 intake. Conversely, copper affects the formation of calcium phosphate crystals and could be involved in the formation of kidney stones, particularly those that contain oxalate. Furthermore, kidney stones featuring trace elements including copper, zinc, and magnesium were shown to be more fragile, which could facilitate their dissolution during extracorporeal shock wave lithotripsy (ESWL).

In conclusion, this study highlights the important role trace metals like copper and magnesium play in managing and preventing urolithiasis. It has been demonstrated that magnesium efficiently prevents the production of calcium oxalate crystals, mainly by restricting the absorption of oxalate from the gastrointestinal system and by decreasing crystal nucleation and growth. This activity provides a promising strategy to lower the risk of kidney stone development, especially when combined with vitamin B6 intake. Conversely, Copper influences the development of calcium phosphate crystals and may play a role in the creation of kidney stones, particularly those that contain oxalate. Furthermore, kidney stones containing trace elements including copper, zinc, and magnesium were shown to be more fragile, which could facilitate their dissolution during extracorporeal shock wave lithotripsy.

Furthermore, plants that are high in magnesium and copper, such Asparagus racemosus, Pedalium murex, and Trachyspermum ammi, show promise as medicines by blocking urolithiasis in a number of ways. In addition to protecting renal epithelial cells, they include their, antioxidant activity, crystal aggregation inhibition and diuretic effects. The bioactive substances in these plants provide a different and intriguing approach to kidney stone management, especially as there are currently few non-invasive therapy alternatives for urolithiasis. More research in clinical settings is necessary to determine whether trace element supplementation and plant-based therapies may prevent and treat urolithiasis in a comprehensive and successful manner.

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Received on 21.01.2025 Revised on 17.03.2025

Accepted on 28.04.2025 Published on 12.07.2025

Available online from July 21, 2025

Asian Journal of Pharmaceutical Analysis. 2025; 15(3):175-180.

DOI: 10.52711/2231-5675.2025.00027

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

MATERIALS AND METHODS41–43