Formulation And Evaluation of the Anti- inflammatory Potential of Shata Dhauta Ghrita enriched Moringa Oleifera Seed Emulgel for Topical Application

Figure No 1 : Acute VS Chronic Inflammation

Herbal topical gel formulation important because they offer localized therapeutic action, better patient compliance, and fewer systemic side effects than oral dose forms, herbal topical gel formulations are significant. Gels have several benefits, including improved skin penetration, quick drug release, ease of spreading, and non-greasy nature. Because herbal extract contain bioactive phytoconstituents such flavonoids, tannins, and alkaloids, adding them to topical gels improve their efficacy in treating inflammation, wound healing, burns, acne, and arthritis. Herbal gels are also generally seen as safer, more affordable, and more acceptable for long- term use^.[5]   Emulgel is a dual controlled drug delivery technology that overcome the drawbacks of traditional topical formulations by incorporating an emulsion (O/W or W/O) into a gel foundation. Emulgel is a new topical drug delivery method that combines the benefits of gels and emulsions, making it particularly useful for delivering both hydrophilic and hydrophobic medications through the skin.^[6][7] Traditional topical formulations, such creams and ointments, have restricted drug release, a greasy texture, and poor anti- inflammatory medication penetration. When used topically, many anti- inflammatory drug are less effective because they are poorly soluble in water. By combining the anti- inflammatory medication in an emulsion system distributed throughout a gel base, Emulgel overcomes these drawbacks and improves skin penetration, prolongs local activity, and minimizes systemic adverse effects, making it an excellent choice for treating localized inflammation.^[8]

MATERIALS AND METHODS :

Shata Dhauta Ghrita 

Ayurveda values ghee for improving intellect, digestion, and body balance, treating various disorders, and supporting modern topical formulations.^[9]  Shata Dhauta Ghrita is an Ayurvedic preparation made by washing cow ghee 100 times with purified water while chanting Vedic mantras. This process produces an odorless, butter-like, silky cream that can penetrate all seven layers of the skin without clogging pores.^[10] Handmade Shata Dhauta Ghrita is an emollient that heals damaged skin by deeply nourishing all tissue layers and is effective for burns, wounds, scars, blemishes, burning sensations, and herpes.^[11]

Figure No 2 : Shata Dhauta Ghrita

Moringa Oleifera Seeds

Moringa oleifera Lam. is a species of the genus Moringa (family Moringaceae)  It is widely grown in India, Pakistan, Afghanistan, Bangladesh, Africa, and Latin America.^[12]

They bacteria Staphylococcus aureus and Pseudomonas aeruginosa, which infect the skin, are effectively treated with moringa seeds^.[13]In Binga District, Zimbabwe, the Tonga people use root powder mixed with milk as an aphrodisiac and traditional remedy. It is believed to help treat asthma, gout, rheumatism, enlarged spleen or liver, digestive gas, stomach disorders, and to relieve ear and tooth pain.^[14][15]

Figure No 3 : Moringa Oleifera seeds

Preparation of Extract :

Cucumber Gel Extract :

Typically, Cucumis sativus L. (C. sativus) is served as a dessert or appetizer.

They are also linked to calming, emollient, cooling, and therapeutic properties. Above all, it is discovered that they have a broad range of activities, including antioxidant^.[16][17].

Three unripe cucumbers were washed, dried, and separated into peel and pulp. Each portion was blended with PBS (1:3), incubated at 37⁰ C for 8 h, filtered, and centrifuged to obtain the supernatant. The extracts were spay-dried using 5% maltodextrin, and aqueous extracts were prepared similarly using deionized water. Phytochemical screening was performed to identify tannins, flavonoids, alkaloids, saponins, and steroids.^[18][19]

Figure No 4 : Cucumber Gel

Lemongrass Oil :

Lemon grass, or Cymbopogon citratus (DC.) Stapf (Poaceae family), is one of the primary aromatic and therapeutic plants grown in Algeria. It  is a perennial tropical grass with thin, long leaves ^[20]

Figure No 5 : Lemongrass Oil

The stalks and leaves of lemongrass were separated, allowed to air dry, and then cut into 4–8 mm pieces. Every extraction used a constant mass of 200 g. Using a Clevenger-type device and distilled water, essential oils were extracted individually by hydro-distillation. Dichloromethane was used to extract the oil, which was then dried over anhydrous sodium sulfate. For additional analysis, the recovered oils were kept at 4–6 °C in the dark.^[21]

Formulation of Emulgel :

Master Formula :

Table No 1 : Batch wise Ingredients

Optimized Formula :

Table No 2 : Optimized Formula Ingredients

Preparation of Emulgel :

1. Preparation of  HPMC Gel Base:

This creates the emulsion's aqueous phase. Alcohols and water are often utilized agents^.[22] HPMC was dispersed in hot, purified water (70°C) to create the gel in formulas F1, F2, and F3. The gel was then chilled for 15 minutes . ^[23][24]

Figure No 6 : HPMC Gel base

2. Preparation of Oil Phase :

Tween 80 was dissolved in purified water to create the aqueous phase of the emulsion, whereas span 60 was dissolved in light liquid paraffin to create the oil phase. Moringa oleifera seed extract was dissolved in ethanol, and methyl and propyl parabens were dissolved in propylene glycol before being combined with the aqueous phase. The oily and aqueous phases were heated independently to 70°C. ^[23][24]

Figure No 7 : Oil Phase

3. Gel preparation :

A mechanical shaker was used to continuously swirl various polymer concentrations in distilled water at a moderate speed in order to prepare the gel bases. Triethanolamine (TEA) was used to bring the pH of each formulation down to 6–6.5. ^[25]

Figure No 8 : Gel Preparation

4. Preparation of Emulgel

The obtained emulsion was mixed with the gel with gentle Stirring to obtain the emulgel^.[25]

Figure No 9 : Gel Formulation

Evaluation of Herbal Emulgel

Visual inspection was used to assess the physical appearance, noting color, homogeneity, uniformity, and phase separation.^[26]

1.Organoleptic properties

Colour :- Pale yellow to Off white (Uniform Appearance and No Visible Discoloration)

Odour :- Characteristics Pleasant Ghee Odour

Appearance :- Semi solid Consistency  Smooth and Glossy Surface No visible particle

Texture:- smooth Soft on touch

Consistency :- Semi solid At room Temperature

Homogenicity :- Appears Uniform No visible particle

2. Evaluation Tests :

Stability Test :                                                                                                                                            For 35 days at room temperature, the final formulations were assessed for their physicochemical stability criteria, including pH, appearance, odor, homogenicity, Spreadability, Phase Separation and washability.^[27]

Dye test :                                                                                                                                                                    Take small amount of emulgel on a clean glass slide. Add few drops of suitable dye, Mix gently with a glass rod and Observe under microscope/ eye.^[28]

Ease of removal test :                                                                                                                                             Apply small quantity of emulgel on skin. Allow it to remain for fixed time (5-10 min), Wash area with tap water. Observe whether formulation remove easily or leaves residue.^[28]

Viscosity Test :                                                                                                                                                               Transfer emulgel into Viscometer beaker by using Brookfield Viscometer and by using suitable spindle at specific rpm. Then record viscosity reading in cps.^[29]    

Spreadability Test :                                                                                                                                                                 Spreadability of the prepared emulgel was determined to evaluate the ease of application on the skin. A known quantity of the formulation was placed between two clean glass slides. A definite weight was placed over the upper slide to form a uniform thin layer. The time required for the upper slide to move a specified distance under the influence of the applied weight was recorded.^[30]                                                                                                                                                                                                                                                                                                                                             

Pharmacological Screening :

In-vitro Anti-inflammatory Activity of Shata Dhauta Ghrita and Moringa oleifera seed extract herbal emulgel by Protein denaturation method

Materials:

Protein Source- Egg albumin/ Bovine albumin                                                                                                    Test Sample- Herbal emulgel (Shata Dhauta Ghrita and Moringa oleifera seed extract)                                              Standard Drug- Diclofenac sodium                                                                                                                        Phosphate Buffer (pH 6.4-6.8)                                                                                                                                Distilled water                                                                                                                                                              Water bath, Centrifuge

Procedure:

1. Preparation of Albumin Solution                                                                                                                          - Prepare 5% egg albumin solution ( 5ml egg albumin + 95ml distilled water                                                      
2. Preparation of Reaction Mixture                                                                                                                                    Divide into 3 groups :                                                                                                                                            A] Control : 2 ml albumin + Phosphate buffer                                                                                                     B] Standard : 2 ml albumin + Diclofenac sodium solution                                                                                  C] Test (Emulgel) : 2 ml albumin + emulgel extract at different concentrations
3. Incubation                                                                                                                                                                   - Incubate all samples at 37⁰ C for 15 minutes
4. Calculation

% Inhibition = (Wc-Wt)/Wc  x 100

Where :                                                                                                                                                                            W_c = Weight of control                                                                                                                                                                                                                                  W_t= Weight of test

50. Control Weight = 2.34
50. Test Weight = 0.40

% Inhibition = 2.34-0.40/2.34 x 100

1.94/2.34 x 100 = 82.9 % 

          The herbal emulgel containing Shata Dhauta Ghrita and Moringa oleifera seed extract showed 82.9 % inhibition of protein denaturation, indicating strong anti- inflammatory activity.[31][32][33]

Evaluation Parameters :

Table No 3 : Evaluation Parameters of Formulation

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      FUTURE SCOPE :

The current study demonstrates a novel method for topical anti-inflammatory use by mixing Shata-Dhauta Ghrita with Moringa oleifera extract in an emulgel system. This innovative composition provides a special fusion of contemporary medicine delivery methods with traditional Ayurvedic principles. To further improve medication penetration, targeted administration, and therapeutic efficacy, future research can investigate the creation of sophisticated carriers such phytosomal systems or nanoemulgels. Furthermore, adding additional synergistic herbal extracts could increase the formulation's overall effectiveness. 

  To confirm its safety and efficacy in human use, more research incorporating in vivo tests and clinical trials is necessary. The bioactive substances causing the observed activity can be identified and standardized using sophisticated analytical techniques. Sustainable development may also take into account stability improvement through the use of natural preservatives and environmentally friendly packaging. Additionally, this formulation has the potential to be expanded into cosmeceutical applications, such as skin healing and anti-aging products, creating new opportunities for herbal therapies innovation and commercialization.   

CONCLUSION

An emulgel combining Shata-Dhauta Ghrita and Moringa oleifera extract for topical application was successfully created and assessed in this study. Good consistency, stability, and compatibility for skin application were demonstrated by the produced formulation's satisfactory physicochemical qualities. An innovative and successful method of anti-inflammatory treatment is provided by combining a classic Ayurvedic basis with a botanical extract. The formulation appears to have promising anti-inflammatory potential, which could be helpful in the treatment of a number of skin-related disorders, according to the observed results.  All things considered, the study is in favor of combining herbal components with contemporary formulation methods to create topical medication delivery systems that are both safe and efficient. With better patient compliance and fewer adverse effects, the emulgel may be a viable substitute for traditional synthetic formulations. To establish its clinical efficacy and investigate its entire therapeutic potential, more research is advised.   An emulgel of Shata-Dhauta Ghrita and Moringa oleifera extract with acceptable physicochemical characteristics that may be used topically was effectively created by the study. The mixture showed encouraging anti-inflammatory capabilities, suggesting that it could be used to control skin inflammation. All things considered, it is a novel, safe, and efficient herbal substitute; nonetheless, additional research is needed for clinical confirmation.

RESULT & DISCUSSION

The physicochemical properties of Shata-Dhauta Ghrita  and Moringa oleifera extract emulgel were good. It was discovered that the preparation had a uniform distribution of the ingredients since it was smooth, homogenious, and grittiness-free. The formulation pH was within an acceptable range for topical administration, indicating good skin compatibility. Appropriate application ease was demonstrated by the spreadability test, which is crucial for patient compliance. Furthermore, during the research period, the formulation showed good stability with no phase separation or notable appearance changes.

The synergistic impact of Moringa oleifera extract and Shata-Dhauta Ghrita is demonstrated by the reported anti-inflammatory action. The findings imply that the topical distribution of herbal components is successfully improved by the emulgel formulation. All things considered, the results validates this formulation’s applicability as a possible herbal substitute for the treatment of inflammatory skin disorders. It advised that more research be done to confirm these findings by clinical and in vivo assessments.

Figure No 10 : Overall Performance Comparison of Formulations