Formulation and Evaluation of Herbal Cream for Treating Psoriasis

 

Padmini Iriventi*, N. Vishal Gupta

Department of Pharmaceutics, JSS College of Pharmacy, Sri Shivarathreeshwara Nagara, Mysuru,
JSS Academy of Higher Education and Research, JSS Medical Institutions Campus,

Sri Shivarathreeshwara Nagara, Mysuru – 570015, Karnataka, India.

*Corresponding Author E-mail: paddhu.iriventi@gmail.com

 

ABSTRACT:

Psoriasis is the most common chronic autoimmune disease. The objective of the present study was to formulate herbal cream that consists of Azadirachta indica (Neem) extract used in treating Psoriasis. Various phytochemical identification tests were carried out using reagents. DPPH (2,2- diphenyl-1-picrylhydrazyl) free radical scavenging assay was carried out to confirm antioxidant activity. Coconut oil, Olive oil and Vitamin E oil were used in the formulation which provides different pharmaceutical activities. Phytoconstituents present in the herbal extract were identified by Liquid Chromatography-Mass Spectroscopy (LC-MS) studies.  Evaluation studies were carried out for prepared cream. LC-MS studies concluded that various herbal constituents were identified and it concludes that active constituents responsible for treating psoriasis were present in the obtained extract and also possessed antioxidant activity.

 

KEYWORDS: Azadirachta indica; Psoriasis; LC-MS; Phytochemicals; Antioxidant activity.

 

 


INTRODUCTION:

Psoriasis is a skin disease which is distinguished by massive proliferation, thick inflammatory cell infiltrates, generation of new blood vessels, modifications in lymphatic structure and impaired differentiation of epidermis. It is an autoimmune disorder where environment and genetic components have a major function. The immune system releases proinflammatory cytokines and growth factors that accelerate the growth of skin cells which accumulate and form thick red patches of skin on various parts of the body1,2.

 

Azadirachta indica (Neem) belongs to the family Meliaceae. Different parts of this tree have various uses and medicinal properties.

 

Neem leaves are useful in treating chickenpox, increase immunity of the body, reduce fever caused by malaria, treating various foot fungi, useful against termites, used in curing neuromuscular pains and in treatment of skin diseases like leprosy, psoriasis etc. Neem seed cake is used as a natural fertilizer and insecticide. Neem bark and roots helps in controlling fleas and ticks on pets, fights against skin infections such as acne, psoriasis, scabies, eczema, etc, treats diabetes, AIDS, cancer, heart disease, herpes, allergies, ulcers, hepatitis and several other diseases3.

 

Pharmacological actions of Neem are Abortifacient, analgesic, anthelminthic, antibacterial, antiyeast, antiulcer, antifilarial, antifungal, antihyperglycemic, anti-inflammatory, antiviral, antimalarial, diuretic, antipyretic, antispasmodic, insecticidal, antispermatogenic, antitumor, hypercholesteremic, hypoglycaemic, immunomodulator.

 

Chemical composition of neem leaves:

The most important bioactive principal are Nimbin, Nimbidin and Azadirachtin. Other compounds present are Nimbidol, Sodium nimbinate, Quercetin, Gedunin, Salannin. In present study methanolic extract of neem leaves is used4.

 

 

MATERIALS AND METHODS:

Azadirachta indica leaves were obtained from Govinda Raj Shetty stores, Mysuru, India. All other reagents used were of analytical grade.

 

Preparation of extracts:

Total Methanolic Neem Extract (MeNE) was prepared by maceration technique. For this purpose, dry leaves (500gm) of Neem were extracted with 1.5 liter methanol and then evaporated by rotary evaporator. The total methanol extract was preserved at 4°C until being analyzed5.

 

·       Qualitative analysis of phytochemicals:

Herbal extract was subjected to preliminary phytochemical screening. Presence of alkaloids (Mayer’s test), flavonoids (alkaline reagent test), tannins (Braymer’s test) carbohydrates (Molischs test), glycosides (Liebermann’s test), saponins (Salkowski test), triterpenoids (Liebermann Burchard test), proteins and amino acids (Ninhydrin test) were tested5-7.

 

·        LC-MS studies:

The herbal extract was analyzed by LC-MS method in order to identify different constituents present in them8. Specifications of Instrument used in the study was as below:

 

Specifications:

LC column: ACQUITY UPLC BEH C18 1.7µm, Solvent selection A: 0.1% Formic acid in water, B: Acetonitrile, Mobile Phase: Water: Methanol, Ionization Mode: ES+, Mode: Positive, Injection Volume: 2 microlitre, Column Dimension: 25cm×2.5mm, Mass range: 50-1500, Software: 1.40.2532.

 

·       DPPH free radical scavenging assay:9

The free radical scavenging activity (antioxidant capacity) of extracts on stable radical 1, 1-diphenyl -2-picrylhydrazyl (DPPH) was estimated. Briefly, 2ml of extract at varying concentrations (50μg/ml to 250μg/ml) was mixed with 2.0ml of DPPH solution in methanol (0.004% w/v). The mixture was allowed to stand at room temperature in dark for 20 min. Then the mixture was vortexed and absorbance was recorded at 517 for neem using spectrophotometer. Ascorbic acid was used as a reference standard and control consisted of DPPH solution without extract. The test was performed in triplicate and percentage scavenging of DPPH free radical by extract was calculated using the equation:

 

(Acontrol- Atest)/Acontrol X 100

 

where Acontrol is the absorbance of control and Atest is the absorbance in presence of extract or standard.

 

 

Preparation of Herbal Cream:10

Oil in water (O/W) emulsion-based cream (semisolid formulation) was formulated. The emulsifier (stearic acid) and other oil soluble components (cocoa butter, cetyl alcohol, coconut oil, olive oil, Vit E oil) were dissolved in the oil phase (Part A) and heated to 75°C. The preservatives and other water-soluble components (methyl paraban, triethanolamine, propylene glycol and ethanol extract of neem) were dissolved in the aqueous phase (Part B) and heated to 75°C. After heating, the aqueous phase was added in portions to the oil phase with continuous stirring and perfume was added. The formula for the cream is given in Table 1.

 

Table 1: Composition of Herbal cream preparation

Ingredients

Quantity

Neem extract (mg)

500

Cocoa butter (g)

3

Stearic acid (%)

3

Cetyl alcohol (%)

2

Coconut oil (ml)

2

Olive oil (ml)

1

Vit E oil (ml)

1

Methyl paraben (%)

1

TEA (ml)

0.5

 

Evaluation of creams:11

·    Type of emulsion under dye test:

The scarlet red dye is mixed with the cream. A drop of the cream was placed on a microscopic slide, then it was covered with a cover slip and examined under a microscope. If the disperse globules appear red and the ground is colorless, the cream is O/W type. The    reverse condition occurs in W/O type cream i.e. the disperse globules appear colorless in  the red ground.

·    Appearance:

The appearance of the cream was judged by its color, pearlscence and roughness and graded.

·    Homogeneity:

The formulations were tested for the homogeneity by visual appearance and by touch.

·    pH of the Cream:

The pH meter was calibrated using standard buffer solution. About 0.5 g of the cream was weighed and dissolved in 50.0 ml of distilled water and its pH was measured.

·    Viscosity:

Viscosity of the formulation was determined by Brookfield Viscometer at 100rpm, using spindle no 7.

 

RESULTS AND DISCUSSION:

Qualitative analysis of phytochemicals:

Different phytochemicals present in both the extracts were identified using methods mentioned in Table 2. Results obtained were also given in Table 2: The results state that prepared herbal extract contained alkaloids, flavonoids, tannins, carbohydrates, glycosides, saponins, triterpenoids.


 

Table 2: Phytochemicals present in Methanolic extracts of Azadirachta

S No

Constituents

Test

Obesrvation

Results

1        

Alkaloids

Mayer’s test

Formation of creamy precipitate.

Positive

2        

Flavanoids

Lead acetate test

Formation of yellow precipitate

Positive

3        

Carbohydrates

Molish’s test

Formation of violet ring at the junction.

Positive

4        

Triterpenoids and steroids

Salwonski test

If lower layer turns red indicates presence of steroids.Golden yellow layer at bottom indicates presence of triterpenoids

Positive

5        

Deoxy sugars

Killer kiliani’s test

Formation of blue color in the acetic acid layer

Positive

6        

Glycosides

Legal’s test

Formation of pink to blood red color

Positive

7        

Reducing sugars

Benedict’s test

Solution appears green or yellow or red depending on the amount of reducing sugar present in the test solution

Positive

8        

Amino acids

Ninhydrin’s test

Formation of blue color

Negative

 


LC-MS studies:

Base peak Ionization (BPI) Chromatogram of methanolic Neem extract was obtained as shown in Fig. 1 which states that retention time of constituent Nimbin (Molecular weight-540.601g/mol) present in the methanolic extract was around 1.62. Fig.2 shows the Mass spectrum related to this RT. Similarly RT of another constituent Azadirachtin (Molecular weight-720.721g/mol) was around 3.16. Its Mass spectrum is shown in Fig. 3.


 

 

Figure 1: Base Peak Ionization (BPI) Chromatogram of Neem

 

 

Figure 2: Mass spectrum of Nimbin

 

 

Figure 3: Mass spectrum of Azadirachtin


DPPH free radical scavenging assay:

The antioxidant activity is mainly depends on phenolic compounds, alkaloids, terpenoids and their derivatives. All these compounds and their derivative were present in the herbal extracts. These compounds produce the free radical and then react with DPPH and gradually changed its colour. In this present study, all these herbal extracts were able to decolorize with DPPH. The DPPH scavenging (%) of Methanolic neem extract was 12.04±1.62%.

 

Evaluation of creams:

·    Dye test:

It confirmed that  formulation prepared  was o/w type emulsion cream

·    Appearance:

There is no change in colour of cream

·    Homogeneity:

By visual appearance and by touch, it is confirmed that the formulation is homogenous

·    pH of the Cream:

The formulation had shown pH nearer to skin,i.e., 6.4

·    Viscosity:

The viscosity of cream was 649 cps which indicates that the cream is easily spreadable by small amounts of shear.

 

Table 3: Evaluation parameters of formulated cream

Parameters

Results

Dye test

o/w type emulsion

Appearance

No change

Homogeneity

Homogenous

pH

6.4

Viscosity (cps)

649

 

CONCLUSION:

From the results obtained it was confirmed that herbal extract of Azadirachta indica prepared, contains various phytoconstituents that provide several medicinal properties. The DPPH free radical scavenging assay performed during the current study confirm the antioxidant property of herbal extracts. LC-MS studies confirm various constituents present in the extract.

 

ACKNOWLEDGEMENT:

Authors are thankful to The Principal, JSS College of Pharmacy, Mysuru, India for providing necessary facilities.

 

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Received on 20.12.2019           Modified on 27.02.2020

Accepted on 18.04.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(1):167-170.

DOI: 10.5958/0974-360X.2021.00029.9