Formulation and Evaluation of Anti-Aging Serum using Ferulic Acid from Matsyakshi Plant

Fig.1 Structure of Skin

OVERVIEW ON SKIN AGING

Aging skin is characterized by various signs, including: Wrinkles, Uneven pigmentation, Darkening, Thinning, Sagging, Roughening. These changes can be attributed to either intrinsic (internal) or extrinsic (external) factors. Theories suggest that DNA damage and repair processes contribute to aging, leading to: Epigenetic changes, Cell senescence, Loss of cell function, Genomic abnormalities. Extrinsic aging is driven by environmental factors like: UV radiation, Pollution, Reactive oxygen species production. Intrinsic aging is linked to: Programmed aging, Cellular senescence, Endogenous oxidative stress, Cellular damage. [5]

Fig.2 Difference between young Skin and Aged Skin

INTRINSIC SKIN AGING

It is a chronologically-driven physiological transformation process. Essentially, intrinsic skin aging is a condition that only appears in old age and is symbolized by smooth, unblemished, pale, dry, less elastic skin that has fine wrinkles. Intrinsic aging is characterized by a decline in dermal fibroblast count, which lowers the synthesis of collagen and elastin, especially types I and III collagen, and results in skin thinning, wrinkles, and loss of elasticity. Skin aging is partly caused by increased expression of matrix metalloproteinases (MMPs), which break down collagen and elastin in the dermal skin layer. Antioxidants such as resveratrol and iso orientin can reduce its expression both intrinsic and extrinsic variables can contribute to skin aging. [6]

Fig.3 Intrinsic factor responsible for skin aging

EXTRINSIC SKIN AGING

Low-grade chronic UVR exposure is Responsible for over 80% of the aging process of Facial skin, while exposure can also result in Sunburn, tanning, inflammation, and damage to Dermal connective tissue. Extrinsically aged skin, Primarily from UVR rays, has sallow complexion with uneven pigmentation, rough texture, coarse Wrinkles, and decreased skin elasticity. Accelerated skin aging can be caused independently by long-term exposure to UV Radiation and cigarette smoking. ROS produced by UV exposure lead to harmful oxidative stress. Because 02 can take electrons, it produces extremely reactive chemical compounds known as ROS. Reactive oxygen species (ROS) are produced when the skin is exposed to photoaging stimuli. These ROS subsequently stimulate dermal enzymes Such as collagenase and elastase. These enzymes Speed up premature skin aging by breaking down Elastin and collagen, respectively. Signs of this Process include deep furrows or severe atrophy, Wrinkles, freckles, sallowness, laxity, and a leathery Appearance. 

Fig.4 Extrinsic factor responsible for skin aging

FUNCTION OF SKIN :

1. Provide a barrier of defence against harmful substances, mechanical, thermal and physical harm.
2. Prevents moisture loss.
3. Minimizes the negative effects of UV light.
4. Aids in temperature regulation.
5. Serves as a sense organ .[9]

FACE SERUM :

Face serums are formulated to deliver a high concentration of active ingredients without unnecessary additives. Their thinner consistency and absence of heavy oils enhance absorption and penetration, making them effective for nourishing and layering on the skin. The focus on essential ingredients without emulsifiers ensures optimal delivery of active agents, such as vitamins or botanical extracts.

ADVANTAGES OF ANTI-AGING SERUM:

1. Keep skin moisturized

2. Help remove skin blemishes

3. Anti-aging benefits and protect against wrinkles [11]

AIM OF PROJECT:

To formulate and evaluate a stable, effective, and cosmetically acceptable anti-aging serum containing selected active ingredients that reduce signs of skin aging such as wrinkles, fine lines, dryness, and loss of elasticity.

NEED OF THE PROJECT:

Degradation of collagen and elastin Increased oxidative stress due to free radicals Reduced skin hydration Loss of skin firmness and glow Formation of wrinkles and fine lines.

OBJECTIVE OF PROJECT :

1. To formulate a stable and cosmetically acceptable anti-aging serum using suitable active ingredients and excipients.
2. To select and incorporate anti-aging agents (e.g., antioxidants, vitamins, humectants, peptides) based on their proven skin benefits.
3. To optimize the formulation for desirable properties such as pH, viscosity, spreadability, and skin feel.
4. To evaluate the prepared serum for physicochemical parameters including appearance, pH, viscosity, homogeneity, and stability.
5. To assess the antioxidant activity of the formulation by suitable in-vitro methods.
6. To study the compatibility of drug and excipients used in the formulation.
7. To perform stability studies of the serum as per ICH guidelines.
8. To evaluate the safety of the formulation by conducting skin irritation tests.
9. To assess the moisturizing and skin-hydration effect of the serum.
10. To compare the prepared formulation with a marketed anti-aging serum.

DRUG PROFILE :

Fig.5 Ferulic acid

Fig. 6 Alternanthera sessilis

Table.1 Drug profile [12]

LITERATURE REVIEW :

1. (Graf, 1992) Antioxidant Activity: Ferulic acid is a naturally occurring phenolic compound that exhibits strong antioxidant activity by scavenging reactive oxygen species (ROS) and preventing oxidative damage to lipids, proteins, and DNA. This property makes it highly useful in topical formulations aimed at protecting the skin from environmental stress.

2. (Lin et al., 2005) Synergistic Effect with Vitamin C: Ferulic acid enhances the antioxidant capacity of Vitamin C and Vitamin E when used in combination. This synergistic effect significantly increases the photoprotective potential of the formulation compared to individual antioxidants.

3. (Pinnell et al., 2001): Stabilization of Vitamin C: L-ascorbic acid is highly unstable and degrades rapidly when exposed to light, heat, and oxygen. Ferulic acid stabilizes Vitamin C by reducing its oxidation, thereby improving formulation stability and shelf life.

4. (Murray et al., 2008): Photoprotective Effect: Topical application of ferulic acid combined with Vitamin C reduces UV-induced erythema, inflammation, and cellular damage. This combination has been shown to significantly improve protection against UV radiation.

5. (Briganti and Picardo, 2003): Anti-aging Properties: Oxidative stress contributes to premature skin aging, including wrinkles and loss of elasticity. Antioxidants such as ferulic acid and Vitamin C reduce these effects by protecting skin structures and improving skin appearance.

6. (Pullar et al., 2017): Collagen Synthesis: Vitamin C plays a key role in collagen synthesis by acting as a cofactor in hydroxylation reactions, while ferulic acid protects collagen fibers from oxidative degradation.

7. (Cavalcanti et al., 2020): Anti-inflammatory Activity: Ferulic acid exhibits anti-inflammatory effects by inhibiting inflammatory mediators, reducing redness and irritation in the skin.

 8. (Saija et al., 1999): Skin Brightening Effect: Ferulic acid inhibits tyrosinase activity, which reduces melanin synthesis and leads to improved skin tone and reduction in hyperpigmentation.

9. (Das and Wong, 2020): Formulation Challenges: Ferulic acid has limited solubility and stability in aqueous systems. Advanced formulation strategies such as nanoencapsulation and emulsions improve its stability and skin delivery.

10. (Telang, 2013): Evaluation Parameter pH: The pH of Vitamin C serum plays a critical role in stability and penetration. Optimal pH (2.5–3.5) enhances absorption and effectiveness.

 11. (Patil et al., 2020): Evaluation Parameter: Drug Content: Quantitative analysis of ferulic acid in formulations is commonly performed using RP-HPLC methods to ensure accuracy, precision, and consistency.

12. (Mancuso et al., 2021): Evaluation Parameter; In-vitro Release: In-vitro diffusion studies demonstrate controlled release of ferulic acid from optimized formulations, improving bioavailability.

13. (Pinnell et al., 2001): Stability Studies: Accelerated stability studies show that ferulic acid improves the stability of Vitamin C formulations under various storage conditions.

14. (Liu et al., 2026): Clinical Evaluation: Clinical studies report improvements in skin hydration, reduction in erythema, and enhanced skin texture following the use of ferulic acid and Vitamin C formulations.

15. (Marcato et al., 2020): Safety and Efficacy: Ferulic acid-based serums are generally safe, non-irritating, and suitable for long-term use in dermatological applications .

PLAN OF WORK :

Aim, need and objective

Selection of ingredients and their roles Extraction, isolation, Purification and Identification of Alternenthra sessilis plant

Formulation and development

Evaluation

Result and Conclusion

MATERIAL AND METHODOLOGY :

Table 2. Master Formula

Preparation of Alternenthra sessilis plant leaf extract :

Method of extraction: Maceration

Take 100g of Matsyakshi leaf powder

↓

after 2 to 3 hour we get powder extract

↓

Transfer the drug to the round bottom flask

↓

extract to the 80% of the ethanol for 48 hours

↓

Filter and Concentrate using rotary evaporator

↓

After 2 to 3 hours we get powder extract

Fig.7 Extraction of ferulic acid from Alternenthra sessilis plant by Maceration

Isolation and purification of ferulic acid :

Procedure :

Dissolve extract in distilled water

Add 2N NaOH

Heat at 60–80°C for 1–2 hours

↓

Cool solution

Add HCl dropwise until pH = 2–3

Result: Ferulic acid precipitates out

↓

Transfer mixture to separatory funnel

Extract with ethyl acetate (3×)

Collect organic layer

Contains: Ferulic acid

↓

Evaporate ethyl acetate using rotary evaporator

Obtain semi-solid crude ferulic acid

Fig.8 Isolation of ferulic acid

Purification:

Method A: Recrystallization

Dissolve in hot ethanol

Cool slowly → yellow crystals form

↓

Method B: Column Chromatography

Stationary phase: Silica gel

Mobile phase: Hexane: Ethyl acetate (gradient) [16]

Identification :

1. TLC Analysis :

Fig.9 TLC Analysis

RF Value = 0.42

Procedure for Anti-aging Serum :

Preparation of gel phase (A)

Disperse Carbopol 940 in purified water and allow hydration for 20-30 minutes

↓

Preparation of antioxidant phase (B)

Dissolve a gel phase in water

Dissolve ascorbic acid in a small amount of water

↓

Preparation of humectant phase (C)

Propylene glycol, glycerin, and sorbitol were stirred.

↓

Preparation of oil phase(D)

mixed almond oil with tween20

↓

Combine phases:

Add phase C to phase A

Add phase B to phase slowly

Add phase D with continuous stirring

↓

Add preservative:

Dissolve sodium benzoate and EDTA, mix well, and then

↓

Add pH adjuster:

Measure the pH

Add triethanolamine dropwise until pH 5-6

↓

Make the final volume:

Add purified water and mix thoroughly [16]

Anti-aging serum :

Fig.10 After mixing all the phases

Fig.11 Anti-aging serum

Formulation Trials :

F1: Rejected due to unstable in room temperature

F2: Reject due to acidic pH

F3: Reject due to improper mixing

F4: Reject because,  it was causing irritation

F5: Stability, pH, homogeneity, no irritation

UV spectroscopy test ferulic acid :

Fig. Test ferulic Acid UV Graph

Observation table :

Table.4 Test ferulic acid UV observation

Actual conc. Test ferulic acid = 14ug/ml

Take 0.12g ferulic acid

EVALUATION PARAMETER FOR PREPARED ANTI-AGING SERUM

1. Physical evaluation for serum :

2. Anti-Oxidant Test :

Fig.13 Graph of anti-oxidant activity

Observation table:

Fig.14 Scavenging activity graph

Observation Table :

Result : Maximum anti-oxidant activity = 60%

3. PH Test :

4. Cyclical temperature study :

RESULT :