INTRODUCTION:

Urological Disorders specially the formation of stone in the urinary system is becoming a threat and area of research due to its prevalence at a wide scale throughout the world. Many factors like change in life style, food habits, family history, obesity etc. are the major cause of the disease. Te formation of stone in the urinary system is classified as urolithiasis meaning the stone or calculi generation in the ureter, gall bladder or any other location beside kidney, where the other class is nephrolithiasis, meaning the formation of stone or pebbles in the kidney¹.

The complication of the disease can be speculated as the urinary system is majorly involved in the regulation of complex chemical levels in our body, involved in the maintenance of acid-base and electrolyte balances and even blood pressure, excretion of metabolic wastes through urine, which drains from kidney to gall baldder with the help of a narrow tubular structure called as ureter and finally through urethra².

The clinical symptoms of the disease are extreme pain in the side, back, below the ribs and spread below the abdomen, fowl smelling urine, fever, vomiting and nausea. Around 75% of the calculi in the body are chemically made up of calcium oxalate and generated as a result of binding of calcium ions with free oxalic acid or oxalate often present in many food articles like spinach, nuts, tea etc. but some stone may be composed of calcium hydroxyl phosphate, magnesium ammonium phosphate, urate and cystine^3,4.

As compare to drugs or medication, the most popular way of treating the disease is surgery or sometimes Shock wave lithotripsy, Extracorporeal Shockwave Lithotripsy are also advisable. Since the availability of anti-urolithic drugs is limited and surgical procedures are even more painful, so there is an urgent need for finding out the effective and patient friendly method for its treatment.

Herbal medicines are gaining its popularity for the treatment of many dreadful diseases including urolithiasis and many plant based medicines are available for the prevention and cure of the disease. The present investigation was focussed on finding out the in-vitro anti-nephrolithiatic activity of leaves extract of costus igneus.

Costus igneus also know as insulin plant or spiral flag and belongs to costaceae family. which is mainly found in tropical Africa south America, Australia, Asia central north and south America. Costus igneus recently establish in India it is cultivated in coastal area, uttarkannada district of Karnataka⁵. Different type of phytoconstituents like saponin, alkaloids, triterpene, flavonoids, steroids and glycoside found in costus igneus⁶. Costus igneus have different type of pharmacological activity such as antidiabetic activity, anti microbial activity⁷, anti inflammatory potential⁸, antioxidant⁹, antiproliferative potential¹⁰, hypolipidemic activity¹¹.

Figure 1: Schematic representation of Anti- urolithic activity¹⁶

MATERIAL AND METHODS:

Calcium Chloride, Sodium oxalate, liquid ammonia, distilled water, methanol, Hydrochloric acid, Tris buffer (pH 6.9), Sulphuric acid and Potassium permanganate. Cystone from the Himalaya Drug Company was used as a standard drug.

Collection, drying and successive solvent extraction of leaves of costus igneus:

The leaves of costus igneus was collected in the month of February 2019 from medicinal garden of Smriti College of Pharmaceutical Education, Indore. The authentication of the plant was done by Dr. S.D. Upadhyaya, Professor & Head, Department of Forestry and Medicinal Plants, J.N.K.V.V, Jabalpur. The leaves were shade dry, shattered and screened with the help of 40 no. mesh. It was soxhlet extracted with petroleum ether for defatting for 4hr at 60^oC. After drying and levigation, the marc was extracted with 70% methanol for 4hr at 85^oC and evaporated to dryness. The percentage yield of methanolic extract was calculated.

Phytochemical Examination of extract:

Phytochemical identification tests was performed in the leaf extract of costus igneus as per the standard methods for the qualitative analysis of Phenolic compounds & tannins, Alkaloids, Saponins, steroids, Proteins & Amino acids, fats and oils, Flavonoids, Terpenoids, Carbohydrates & glycosides^12,13.

In-Vitro anti-urolithic activity:

The precipitates of calcium oxalate was prepared by reacting the equimolar solution of calcium chloride dehydrate (Prepared in distilled water) with sodium oxalate (Prepared in 10ml of 2N H₂SO₄. The precipitates was washed with ammonia solution to make them free of H₂SO₄ and finally washed with water and allowed to dry at 60⁰C. The egg membrane was isolated from egg by the use of 1M HCl and 10mg of the leaves extract of costus igneus was packed along with 1 mg. of calcium oxalate precipitates, inside the egg membrane. The membrane was tied on the glass rod and suspended in 100ml. 0.1 M Tris buffer.

The anti-urolithic activity was performed as shown in the table 1.

Table: 1 Group division for in-vitro anti-urolithic activity

Group	Content in semi permeable egg membrane
I (Blank)	1 mg. calcium oxalate
II (Positive Control)	1 mg. calcium oxalate + 10 mg. standard drug (Cystone)
III (Test)	1 mg. calcium oxalate + 10 mg. test drug (Methanolic extract of leaves of costus igneus)

The flasks were incubated for 2 hours at 37⁰C. After 2 hrs. the content from each egg membrane was taken separately and added 2ml. 0.9494 N H₂SO₄ and titrated with 0.1 N potassium permanganate. The pink color indicates the end point and with the help of the burette reading the amount of undissolved calcium oxalate in the egg membrane was calculated and percentage of dissolution was determined^14,17.

RESULTS:

Analysis of pharmacognostic activities of leaves of Costus igneus:

Table 2: Organoleptic features of dried leaves of Costus igneus

Common Name of plant

Scientific

name

Part of the Plant Used in the study

Colour

Insulin plant

Costus igneus

Leaves

Greenish Yellow

Table 3: Color and percentage yield of methanolic extract of leaves of Costus igneus

Name of Extract	Percentage Yield
Methanolic Extract	9.57

Table 4: Phytochemical qualitative tests of extract

Phytochemical tests	Result
Carbohydrates	-
Tannins	-
Alkaloids	+
Glycosides	+
Phenolic compounds	+
Proteins & Amino acids	-
Flavonoids	+
Phytosterols	+
Terpenoids	+
Gums	-
Saponins	+

Table 5: Percentage of Dissolution of calcium oxalate by Methanolic extract of leaves of costus igneous and standard drug: Cystone

S. No	Groups	% of dissolution of Calcium Oxalate
1	Blank	0.00
2	Positive Control	86.12
3	Methanolic Extract	80.34

Figure-2 Percentage of Dissolution of calcium oxalate by Methanolic extract of leaves of costus igneus and standard drug: Cystone

In the present study methanolic extract was obtained from the leaves of Costus igneus with percentage yield of 9.57.

The phytochemical screening of the extract shows the presence of Carbohydrates, Tannins, Alkaloids, Phenolic compounds, Flavonoids, Terpenoids, Saponins, Steroids (Table-4).

The percentage dissolution of methanolic extract and standard drug cystone was found to be 86.12% and 80.34% respectively.

DISCUSSION:

In the present study evaluation of anti urolithic activity of methanolic extract of costus igneus was performed. From the study it was found that the extract shows percentage of dissolution of calcium oxalate but it was less in comparison to standard drug cystone. The methanolic extract shows the presence of various phyoconstituents like flavonoids, phenolic compounds, terpenoids etc. which may be responsible for the anti-urolithic activity.

CONCLUSION:

Lithiasis (stone formation) is an one of the major cause of renal failure. It includes nephrolithiasis i.e calculi formation in kidney and urolithiasis stone or pebble formation in ureter or bladder. The calcium stones are maximally found in men and the occurrence of phosphate stones is more in women¹⁵. The herbal methanolic extract of leaves of costus igneus was explored for its in-vitro anti-urolithiatic activity. The promising results obtained in the present investigation suggests that the plant have anti urolithic activity. The plant leaves contains many important phytochemical which may be responsible for the activity, but further chemical investigation is needed to get complete profile of the phyochemical responsible for the activity.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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Received on 13.08.2019 Modified on 30.10.2019

Research J. Pharm. and Tech 2020; 13(3):1289-1292.

DOI: 10.5958/0974-360X.2020.00237.1