1. INTRODUCTION:

Phyllanthus urinaria is commonly called as chamber bitter, is an herb species in the family which belong in Euphorbiaceae and Phyllantaceaceae family^1,2. Numerous small green red fruits are found along the underside of its stem. Therefore, the plant is called “Ye Xia Zhu” in the Pharmacopeia of P.R of China¹. It has features of great number of seeds, high shade tolerance, and extensive root system and therefore considered a competitive weed.

The whole plant could be used as medicine. As a natural product P. urinaria has long been used in traditional oriental medicine to promote healthy elimination of gallstones and kidney stones, as an immune system inducer and for liver disease treatment^3,4.

Due to the existence of diverse active ingredients, effortlessness of access, and minimal side effects, some medicinal plant extracts and it’s Phytoconstituents are recognized as a hopeful another for antimicrobial and antifungal agents. Medicinal plants with antimicrobial, antioxidant and anti-inﬂammatory properties. Polyherbal results in cheaper medication by reducing the duration of therapy for anti-inﬂammatory and antimicrobial and antifungal medications. Modern medical science, despite so many achievements and progress, is finding itself difficult to reach to every people and deal with ever increasing diseases and disorders⁵. Still, majority of world population mainly in developing and underdeveloped countries does not have access to modern medicine and depends on the time tested traditional or complimentary system of medicine, many of these systems is much older compared to the allopathic medical wisdom⁶.

The whole plant, roots, fruits, and leaves of P. urinaria is used to treatment various complications in different regions of the world. In particular, the Chinese and Indian traditional medicine system documents different applications of parts of this plant as remedies for various health complications. It is used in treatment of jaundice, enteritis, diarrhea, dropsy. Pharmacological activities of P. urinaria extract shows the anticancer, hepatoprotective, antioxidant, anti-diabetic, antimicrobial, antifungal effect⁷.

2. MATERIAL AND METHOD:

The extract was obtained by stem, leaves of Phyllanthus urinaria which is easily available in road side. The preparation of cream is given bellow in steps.

2.1. Plant collection:

Leaves from Phyllanthus urinaria plant were collected from Sangli district in Maharashtra, India. The plant authentication is carried out by botanical department of Yashvantrao Chavan college, Karad, Maharashtra, India. The extraction was separately carried out for different selected plants.

2.2. Preparation of plant extract:

The fresh leaves were washed with running tap water in 15 minutes and dry in show at room temperature for week. The leaves are cut in small pieces then make fine powder of it. 20g of leaves are weighted and dissolved in 100ml distilled water in a 500ml Erlenmeyer flask and boiled for 30 mins. The extract was filtered with glass cloth again filtered with Whatman filter paper number 1 were stored in an air tight container and protected from sunlight for further use⁸.

2.3. Preparation of cream from extract of Phyllanthus urinaria.:

For the preparation of cream from extract there were need of prepare two phases which are given in bellow⁹:

2.3.1. Phase 1: Preparation of oil phase:

Steric acid (17%), potassium hydroxide (0.5%), sodium carbonate (0.5%) was taken into one porcelain dish and this mixture was melted at 70⁰C.

2.3.2. Phase 2: Preparation of aqueous phase:

Alcoholic extract of crude drugs mentioned in 2.2 (4.5%), glycerin (6%), water (71%) were taken into another porcelain dis and heated this mixture at 70⁰C.

2.3.3. Addition of aqueous phase to oil phase:

The aqueous phase was added to oil phase with continuous stirring at 70⁰C. Now, once the transfer was completed it was allowed to come at room temperature, all the while being stirred. Perfume (0.5%) was added at last just before the finished product was transferred to suitable container. Then cream was evaluated for various physical parameters.

3. CHARACTERIZATION OF CREAM:

3.1. Physical characteristics:

The prepared P. urinaria cream is evaluated for its physical properties¹⁰.

3.1.1. pH of the formulated cream:

The pH meter was used to calculate the pH of the formulated cream, and it has been tested with a standardized buffer solution. The pH of 0.5mol of cream were determined after it was calculated and dissolved in 50.0ml of sterile distilled water.

3.1.2. Homogeneity:

The formulated cream was tested sense of touch experiments have been used to assess homogeneity. And ranked it as excellent, very good, good, poor¹⁰.

3.1.3. Consistency:

Consistency is measured by attaching cone to holding rod which was dropped from fixed distance of 10cm. it should be fall on the center of cylinder filled with cream. The distance travelled by cone is measured after 10sec.

3.2. Rheological properties.

3.2.1. Spreadability.

The excess cream was applied in between two glass slides and compressed to uniform thickness by placing 100gm weight for 5min. weight was added to the pan. The time required to separate the two slides, i.e. the time in which the upper glass slide moved over the lower plate was taken measure of spreadability¹⁰.

S = m * l\t

m= weight tide to upper slide

l= length moved on the glass slide

t= time taken.

3.2.2. Viscosity.

A Brookfield synchroelectric viscometer, was used to measure the viscosity (in cps) of P. urinaria cream. The spindle was rotated at 2.5rpm. Sample of the creams were allowed to settle over 30 min at the temperature of test (25⁰C) before the measurements were taken.

3.3. Other properties:

3.3.1. Appearance:

The appearance of the cream formulations was judged based on its pearlscence, color, roughness and uniformity.

3.3.2. Type of smear:

the formulation of smear after applying to skin is checked. A fixed amount of different cream formulations was applied on to the surface of the dorsal human skin and the kind of smear or film the formation on the skin was examined.

3.3.3. Removal:

Applied cream was observing for removal by using tap water. All three creams were applied on the surface of the dorsal skin. The simplicity of removal of the creams applied on the surface was examined by subjecting the applied part for washing under laminar flow of tap water for 10 to 15s.

3.3.4. Irritancy test:

Apply cream at back side of left hand. An area was marked about 1sq.cm on the left-hand dorsal surface. The cream was spread to the skin in the designated region, and the timer was started. The cream application area was checked for Irritancy, edema, erythema if any, atusualinterval up to 24 hrs.

3.3.5. Stability test:

Stability checking at a quicker speed for the cream formulations was conducted for 6 days at room temperature. And for the two most stable cream formulations found during room temperature studies, the accelerated studied were conducted at 40^oC ± 1^oC for 20 days.

Table no 3.1: Physical evaluation result of cream formulations

Sr. No	Parameter	Formulation
1	pH	6.1
2	Homogeneity	Good
3	Consistency	5mm

Table no 3.2. Rheological evaluation result of cream

Sr. No.	Parameter	Formulation
1	Spreadablity	34 g.cm/sec
2	viscosity	10 X 10⁶

Table no 3.3. Other evaluation result of cream

Sr. No.	Parameter	Formulation
1	Appearance	No change
2	Type of smear	Non greasy
3	Removal	Easy removal
4	Irritancy test	Nil
5	Stability test	Good spread

4. PHYTOCHEMICAL CONSTITUENTS OF PLANT EXTRACT:

The phytochemical screening was done for ethanolic extracts showed presence of phytoconstituents alkaloids, flavonoids, tannins, phenolic, terpenoids, reducing sugars and steroids. shown in table (4)¹¹.

Table no 4: Shows the phytochemical constituents of plant extract:

Active Constituents	Test	Test	Present test
Alkaloids	Wagner reagent	Dragendorff’s	+
Flavonoids	Sulphuric acid	NaoH	+
Tannin	FeCl₃	Dil.HNO₃	+
Glycoside	Killer Kilani	Saponin	+
Reducing sugar	Fehling’s test	Benedict’s reagent	+

5. ANTIFUNGAL ACTIVITY:

The antifungal activity is performed by the agar plate method. The potato dextrose agar is prepared with nutrients (6gm), heated water (100ml) and mix the nutrient agar in it. Properly autoclave the all petri plates media. Pour the agar in plates in aseptic area. The striking of fungus (C. albicans and A. niger) on agar plate by the inoculating loop in an aseptic condition. The plates are placed in autoclave of 37⁰C for 24 hrs.

When the growth of fungus is observed the further procedure is done by the well-diffusion method. Making 3 bores with help of borer. The formulation, extraction and standard luliconazole (s) is with micro.gm/ml concentration and place in bore. The plates put in autoclave with no disturbance about 24 hrs. the zone of inhibition is measured in mm. shown in table below table (5). ¹²

Table no. 5: Zone of inhibition in mm measured in petri plate.

Sr.No.	Name of fungus	Zone of inhibition in mm.
		Formulation (100 micro ml)	Extraction (100 micro ml)	Luliconazole (s) (100 micro ml)
1	C. albicans	12.6	13.8	15
2	A. niger	10.5	11.2	12

RESULTS:

The P. urinaria cream is prepared. its evaluation as physical, rheological, and other evaluation test are carried out with good result. Evaluation test results are shown in the table respectively as table no 3.1, 3.2, 3.3.

The phytochemical constituents of plant extract are determined with various chemical reaction. The active constituents found in extract are alkaloid, flavonoid, tannin, glycoside, reducing sugar etc. test and its result shown in table no 4.

The antifungal activity of extract and formulated cream is done on the agar plate by well diffusion and striking method. The zone of inhibition is measured with help of scale the obtained result shown in the table no 5.

DISCUSSION:

In the current study of P. urinaria extract cream is done with formulation of O/W type of cream. The result obtained and confirmed. Cream formulation was easily wash with tap water, spreadable, non-greasy and thin, feel good on applied to skin. The observation gives no irritation on the skin after application. Appearance of formulated cream is stable. The better results of cream are obtained with parameter like homogeneity, spreadability, type of smear, removal, and stability, pH. The antifungal activity of cream, extract is gives the comparative study with standard luliconazole. The extract shows more activity than the cream but the both cream and extract shows antifungal activity.

CONCLUSION:

Prepared cream of P. urinaria have good evaluation test result so it is usable. It does not show any irritation so easily applicable. The removal of cream is good so it easily washed with water. It has many active constituents which are helpful for skin. The main purpose of cream is antifungal action, the antifungal activity of prepared cream is good. The quality of cream as antifungal is checked by carried out antifungal test.

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Received on 28.04.2021 Modified on 14.05.2021

Asian J. Research Chem. 2021; 14(5):353-356.

DOI: 10.52711/0974-4150.2021.00060