Design, Optimization and Evaluation of Nail Lacquer for Treatment of Onychomycosis
Abna Fathima.V, Fawas.V, Greeshma Ann Varghese, Riya Shaji K.M, Shahana Sherin.K,
Tina Raju, Dr. Lal Prasanth ML
Dr. Moopen’s College of Pharmacy, Naseera Nagar, Meppadi P.O., Wayanad, India - 673577.
*Corresponding Author E-mail: tinarcpmht@gmail.com
ABSTRACT:
Nail lacquers or nail paints may be defined as viscous or semi-liquid preparations that are intended for the decoration of the nails of the fingers and toes. Nail lacquers form the most commonly used the most popular type of manicure preparations. It can help our nails grow longer and stronger. It has become an essential step for many self-care routines. An attempt was made to prepare nail lacquer containing natural antifungal agent obtained from extract of pomegranate (Punica granatum) peel which are employed for the treatment of onychomycosis, a fungal infection of nail. This was expected to improve chemical efficacy and improve patient compliance. The film forming polymer, permeation enhancer and solvent, along with extract was optimized for component release, stability and convenience of application. The formulation was prepared by simple mixing and using magnetic stirrer which was analyzed for nonvolatile content, gloss, smoothness to flow, drying time, viscosity, adhesion, permeation studies and antifungal studies. Among all the formulations of nail lacquer prepared, F5 exhibited good nonvolatile content and zone of inhibition. Pomegranate peel extract nail lacquer can be used irrespective of the sex of patient for the treatment of onychomycosis.
KEYWORDS: Nail lacquer, Nail, Onychomycosis, Fungal infection, Treatment, Pomegranate Peel Extract, Formulation optimization, Antifungal activity, Permeation.
The word cosmetics derived from the Greek word “kosmeticos” which means to “adorn”. One of the first cosmetic procedure was getting tattoos and also there where treatments for wrinkled skin. Cosmetics are the preparations that can be used for cleaning, beautifying, conditioning and for protecting the skin, hair, nails, and lips etc. Egyptians were the first one to identify the health benefits of cosmetics.
Nowadays cosmetics have become an essential commodity of life. Cosmetics not only play an important role in beautifying but also in psychological factors like confidence also protect the body from infections and other harmful substances. India is among one of the emerging countries with increased growth in cosmetic industries. In 2009 above 350 billion rupees were registered by the Indian cosmetic industries. So, cosmetics play an important role in day today life and are used widely without any gender.
In humans, most primates, and some other mammals, a nail is a horn-like envelope that covers the dorsal surface of the terminal phalanges of fingers and toes. Fingernails and toenails are made of a tough protein called keratin. The nail is the largest skin appendage. It grows continuously through life in a non- cyclical manner; its growth is not hormone-dependent. The nail of the middle finger of the dominant hand grows fastest with approximately 0.1mm/day. The nail consists of the nail plate, the nail matrix and the nail bed below it, and the grooves surrounding it.
The tissue on which the nail rests is known as the matrix (matrix unguis, keratogenous membrane, nail matrix, or onychostroma). It is a region of the nail bed that extends below the nail root and is home to nerves, lymph nodes, and blood vessels. The cells that develop into the nail plate are produced by the matrix. The fingertip's shape dictates whether the nail plate is flat, arched, or hooked, whereas the size, length, and thickness of the matrix influence the width and thickness of the nail plate. As long as it obtains nutrition and is in good health, the matrix will continue to expand. Round and white new nail plate cells arise from the matrix during incubation, pushing out the older nail plate cells and in this way yet older cells become compressed, flat, and translucent, making the pink colour of the capillaries in the nail bed below visible.
The lunula (occasionally called simply "the moon") is the visible part of the matrix, the whitish crescent-shaped base of the visible nail. The lunula is largest in the thumb and often absent in the little finger.
The skin layer underneath the nail plate is called the nail bed. Like all skin, it is made up of two different tissue types: the superficial epidermis, which is the layer just beneath the nail plate and moves with the plate, and the deeper dermis, which is the living tissue attached to the bone and contains capillaries and glands. The dermis and epidermis are connected by minute longitudinal "grooves" called matrix crests or nail matrix crests. The plate becomes thinner as we age, and these ridges are seen in the plate itself. The deep groove into which the nail root is put is called the nail sinus. The portion of the nail that is located in the nail sinus, or the base of the nail that is buried beneath the skin, is known as the nail root. It comes from the matrix, the below-surface tissue that is actively growing.
The real nail, also known as the nail plate or corpus unguis, is comprised of translucent keratin protein, which is an amino acid-based protein. It creates a strong, flexible substance inside the nail from several layers of flattened, dead cells. The capillaries beneath the plate give the impression that it is pink. The geometry of the underlying bone affects its transverse shape.
The free margin or distal edge is the anterior margin of the nail plate corresponding to the abrasive or cutting edge of the nail. The hyponychium is the epithelium located beneath the nail plate at the junction between the free edge and the skin of the fingertip. It forms a seal that protects the nail bed. The onychodermal band is the seal between the nail plate and the hyponychium. It is found just under the free edge, in that portion of the nail where the nail bed ends and can be recognized by its glassy, greyish colour. It is not perceptible in some individuals while it is highly prominent on others.
The cuticle and eponychium work together to provide a seal of protection. The eponychium is the fold of skin cells that generates the cuticle, and the cuticle is the semi- circular layer of nearly invisible dead skin cells that "ride out on" and cover the rear of the visible nail plate. The eponychium is the small band of epithelium that extends from the posterior nail wall onto the base of the nail. The perionyx is the projecting edge of the eponychium covering the proximal strip of the lunula.
The nail wall is the cutaneous fold overlapping the sides and proximal end of the nail. The lateral margin is lying beneath the nail wall on the sides of the nail and the nail groove or fold are the cutaneous slits into which the lateral margins are embedded. The paronychium is the border tissue around the nail and paronychia is an infection in this area.
Fig. 1: Anatomy of normal and diseased nail
Onychomycosis is a fungal nail disease caused by dermatophytes, non- dermatophytes and yeast which affects nail bed, matrix and plate. The term is from Greek words “onyx” means nail and “mykes” means fungus. This is an infection that makes one feel embarrassed socially as it is characterized by patches or white or yellow discoloration, nails become ragged, brittle or crumbly, thickening of nails, separation of the nail from the nail bed.
The nail as an anatomic structure protects the terminal phalanx of the digit from injury. Historically, it has served as a tool for protection and for survival. Today, there are many nail cosmetic products available, including nail hardeners, nail polishes, nail extensions, artificial/sculpted nails, nail lacquers and nail decorations.
Nail lacquer is derived from the French word “lacre” meaning a kind of sealing wax. It is a type of nail polish or varnish which when applied on the nail will quickly dries off.
The emergence of nail polish can be traced back to ancient China, where it was worn as a symbol of wealth and power by rulers and members of high society. Early nail polish was a mixture of beeswax, gelatin, and egg white dyed with orchids or roses that was applied to the nails and left on for a number of hours, leaving a stain. According to ancient manuscripts, reds and blacks were popular colours, were mostly metallic dusts of silver and gold. It was a crime for anyone else to wear stained nails as they were considered a symbol of the wealthy and powerful.
The ancient Chinese techniques then spread to India, Africa, and the Middle East. Egyptians began using henna to colour their nails. Cleopatra used plant extracts to dye her nails a deep blood red, and henna stained nails were discovered on certain mummified Pharaohs. It was fashionable for women in India and Africa to decorate their fingertips with henna.
Nail polish didn't arrive in Europe until much later, in the late 18th century, until the trade dealers from India and the Middle East reaches there. It was still associated with the wealthy at this point, until the first nail salon opened in Paris in the late nineteenth century. However, nail polish had not yet evolved into a liquid form. Powders and oils were applied to the nails and buffed away, leaving a soft pink or red colour.
Only after nail polish made its way to the United States were advances in nail technology like today's offerings developed. A woman named Mary Cobb opened the first nail salon in the United States in Manhattan in 1878. Her $1.25 buff and shine manicures established her as a successful female entrepreneur.
At this point in the vast history of nail polish, the invention of modern-day nail polish was not far off. During World War I, the United States seized
German chemical patents, allowing Nitrocellulose to enter the American market. In the 1920s, French makeup artist Michelle Menard, who was working for the Charles Revson Company at the time release the first liquid nail polish.
Nail technologies advanced throughout the 1950s, with the introduction of nail wraps and acrylic nails. A dentist accidentally invented acrylic nail. The materials used to fill cavities in teeth were transformed into false nails.
Fast forward to today, and there are virtually limitless options for adding colour to our nails. There is a wide range of products available to suit every individual's desires and aesthetic. At one end of the spectrum are peel-off nail polishes that only last a few hours on the nails, and at the other end of the spectrum are acrylic nails, which allow clients to have any shape, length, and colour they want for several weeks without chipping.
The pomegranate is native to a region from modern-day Iran to northern India. Pomegranates have been cultivated throughout the Middle East, South Asia, and Mediterranean region for several millennia, and it is also cultivated in the Central Valley of California and in Arizona.
1. Phenolic acids
· Hydroxycinnamic acid
· Hydroxybenzoic acid
2. Flavonoids
· Anthocyanin
· Catechin
3. Other complex flavonoids
4. Hydrolyzable tannins
· Ellagic acid
· Gallic acid
· Pedunculagin
· Punicalin
· Punicalagin
· Regenerating skin cells
· Hydrates skin
· Detoxifying the skin, reduce premature aging and hyperpigmentation.
· Protecting from the UV rays.
· Useful for oily, congested skin or excessively dry skin.
· It helps brighten our skin tone and whitens skin.
· Peel has anti-inflammatory, antimutagenic and anticancer activities.
Table no. 1: List of Materials
|
Sr. No |
Name of chemicals |
Supplied by |
|
1. |
Concentrated nitric acid |
CHEMIND Chemicals |
|
2. |
Concentrated sulphuric acid |
CHEMIND Chemicals |
|
3. |
Glycerol anhydrous |
CHEMIND Chemicals |
|
4. |
Hydrogen peroxide |
CHEMIND Chemicals |
|
5. |
Nitrocellulose |
LABCO Scientific |
|
6. |
Sodium hydrogen carbonate |
CHEMIND Chemicals |
|
7. |
Ethyl cellulose |
Research Lab |
|
8. |
Salicylic acid |
NICE Chemicals |
|
9. |
Ethyl acetate |
NICE Chemicals |
|
10. |
Dibutyl phthalate |
LABCO Scientific |
|
11. |
Acetone |
NICE Chemicals |
|
12. |
Potato dextrose agar |
HIMEDIA |
Extraction of Pomegranate Peel Using Maceration:
· Pomegranate peel was collected and cut into small pieces.
· It was then cleaned, washed using distilled water and dried in hot air oven at 400 C for 48 hours.
· The dried powder was grinded to fine powder form.
· 30g of pomegranate peel powder was taken in three iodine flask and 70ml of ethanol was added.
· It was shaken at 6 hours’ interval for 7 days.
· Then it was filtered by using muslin cloth.
· The filtrate was collected.
· Then it was poured in a petri dish to remove alcohol.
· It was kept for one day, so that the evaporation of alcohol takes place.
· Finally, the semisolid pomegranate peel extract was collected.
The formulation of pomegranate peel extract based nail lacquer was done according to the formula prescribed in Table. No.2:
Table. No.2: Formulation of nail lacquer
|
Ingredients |
Formulation code |
||||
|
F1 |
F2 |
F3 |
F4 |
F5 |
|
|
Pomegranate peel extract |
2.5% |
5% |
7.5% |
10% |
12.5% |
|
Nitrocellulose |
2% |
2% |
2% |
2% |
2% |
|
Ethyl cellulose |
1% |
1% |
1% |
1% |
1% |
|
Salicylic acid |
5% |
10% |
15% |
20% |
25% |
|
Acetone |
10% |
10% |
10% |
10% |
10% |
|
Dibutyl phthalate |
0.2% |
0.2% |
0.2% |
0.2% |
0.2% |
|
Ethyl acetate q.s. |
100% |
100% |
100% |
100% |
100% |
i. Nitrocellulose and ethyl cellulose were dissolved in sufficient quantity of ethyl acetate to get clear solution.
ii. Salicylic acid was dissolved in above mixtures and dibutyl phthalate was added.
iii. The extract and acetone were added with continuous stirring at 100 rpm on magnetic stirrer.
iv. The formulations were coded as F1 to F5.
v. Finally, ethyl acetate was added to get proper consistency.
Physicochemical evaluation of Pomegranate peel Extract nail Lacquer11,12,13,14
Organoleptic test was carried out to observe the physical appearance of the preparation using the parameters colour, odor and consistency of the preparation.
1ml of the pomegranate peel extract was taken and diluted with distilled water. Add 1 or 2 drops of ferric chloride. The formation of blackish blue colour confirms presence of tannin.
2ml of sample was taken in a watch glass and initial weights were recorded. Then the watch glass was kept in the oven at 1050C for 1hr, and it is then removed, cooled and reweighed. The difference in weights was recorded. The difference in weight was recorded.
With the help of a brush a film of sample was applied on a petridish. The time required to form a dry-to- touch film was noted with the help of stop watch.
The sample was poured from a height of 1.5 inches into a glass plate and it was then spread on a glass plate and made to rise vertically. Smooth flow of the film was visually observed.
Sample of nail lacquer was applied over the nail and gloss was visually seen and compared with cosmetic nail lacquer.
The viscosity of nail lacquer was determined using Brookfield viscometer. The sample was placed in a Brookfield viscometer. Then 1.5 RPM is selected for the spindle no. 4 to determine viscosity of the formulated batches of nail lacquer. The Brookfield viscometer determines viscosity by measuring the force to turn the spindle in the solution at a given rate.
The nail lacquer was applied to the nail. It was kept for drying and the adhesive (transparent cello tape) was adhered to the nail, and then it was pulled apart. The cello tape was observed for the presence of any lacquer material.
i) Antifungal study of pomegranate peel extraction-
The antifungal activity of the pomegranate peel extract was measured using cup plate method in potato dextrose agar medium.
Table.no. 3: Formula for potato dextrose agar medium
|
Ingredients |
For 100 ml |
|
Potato extract |
0.4gm |
|
Dextrose |
2gm |
|
Agar-agar |
1.5gm |
|
Distilled water |
Up to 100ml |
The zone of inhibition obtained for pomegranate peel extract was recorded. Inoculum of C. albicans was prepared by transferring a colony to potato dextrose broth which was sterilized using autoclave at 121°C for 15min at 15 psi pressure.
Sterile petri dish was taken and potato dextrose agar medium was poured into the petri plate to fill ¾th of its volume. The agar was allowed to cool. 0.1ml of the inoculum was placed at the Centre of the agar plate and spread using sterile L-spreader. The inoculums were allowed to diffuse, the cups of about 1mm size were made using sterile corn borer. The cups were marked with the batch code. About I ml of sample was poured into the cups for each batch and kept for sometimes. These plates were then covered and kept in an inverted position in the incubator at 37°C for 24-48hours. The zone of inhibition was observed after incubation period.
The antifungal activity of the formulated batches of nail lacquer was measured using cup plate method in potato dextrose agar medium. The zone of inhibition obtained for all the batches were compared with the zone obtained for pomegranate peel extract.
Cloven hooves from freshly slaughtered goat were collected from the nearby slaughter house. These were then soaked in phosphate buffer for 24 hours to remove adhering connective and cartilaginous tissue. Membranes with a thickness of about 1mm were cut from the distal part of the cloven hooves. The Franz diffusion cell was used for in vitro permeation studies, and the hoof membrane was carefully placed on the cell. The nail lacquer of each batch were then applied evenly to the nail membrane’s surface. The receptor compartment was filled with a pH 7.4 phosphate buffer solution, and the entire assembly was kept at 37° C with constant stirring. 1 ml sample from receptor compartment was taken at 30 min. This was then tested for antifungal activity using disk diffusion method.
RESULT AND DISCUSSION:
Organoleptic test was carried out to observe the physical appearance of the preparation using the parameters colour, odor and consistency of the preparation. It was observed that as the quantity of pomegranate peel extract increases the colour of the formulation intensifies and was uniformly distributed. The formulation was observed to have a characteristic smell of acetone. The consistency of all batches was found to be proper by comparing with standard marketed nail lacquer formulation.
Fig.2: Pomegranate peel extract nail lacquer
Test for tannins was performed in pomegranate peel extract. A bluish black colour is formed which indicates the presence of tannins.
Fig. 3: Estimation of tannin
Nonvolatile content of F1 to F5 was observed. 2 ml of sample was taken in a watch glass and was spread to an optimum thickness further, initial weights were recorded. Then the watch glass was kept in the oven at 1050C for 1hour, and it is then removed, cooled and reweighed.
The difference in weights was recorded.
W1 -Weight of empty dish
W2- Weight of sample before test
W3- Weight of dish with dried sample
|
Sl. no |
Batch |
Initial weight (W1) |
Final weight (W2) |
Difference (W3) |
Non-volatile content (%) |
|
1. |
F1 |
18.20 |
20.2 |
19.90 |
8.415 |
|
2. |
F2 |
18.52 |
20.52 |
20.00 |
7.212 |
|
3. |
F3 |
16.60 |
18.60 |
17.50 |
4.838 |
|
4. |
F4 |
16.91 |
18.91 |
17.83 |
4.865 |
|
5. |
F5 |
18.39 |
20.39 |
19.10 |
3.482 |
The drying time of different formulation of nail lacquer was noted. The drying time of batches F1, F2, F3, F4, and F5 was found to be 95sec, 90 sec, 83sec, 82sec, and 80sec respectively. From this it can concluded that batch F5 shows lower drying time as compared to other batches.
Table No.5: Drying time
|
Batch |
Drying time (Seconds) |
|
F1 |
95 sec |
|
F2 |
90 sec |
|
F3 |
83 sec |
|
F4 |
82 sec |
|
F5 |
80 sec |
The prepared formulation of nail lacquer was found to be satisfactory with the smoothness to flow. The F1, F2, F3, F4, F5 batches of nail lacquer formulation was poured separately into a watch glass and was found that each batches spread uniformly and resulted in a smooth uniform film.
The gloss of the nail lacquer was observed. The sample of nail lacquer were applied over the nail. It showed gloss and shiny appearance as shown in the figure below.
The viscosity of nail lacquer was determined using Brookfield viscometer. The spindle number 4 was used to determine the viscosity and the temperature was set at 23.7 degree Celsius. The instrument was set up at an rpm of 1.5 to determine the viscosity.
Table. No: 6: Viscosity
|
Batches |
Viscosity (Cp) |
|
F1 |
353298 |
|
F2 |
354942 |
|
F3 |
362562 |
|
F4 |
362881 |
|
F5 |
363793 |
It was found that the formulated batches almost show same viscosity. And compared to other batches F5 offers sufficient viscosity.
The adhesive material was observed for presence of any lacquer material. We saw that there was no lacquer film residue on cello tape after removing, hence it was found that the prepared nail lacquer has good adhesive property.
The zone of inhibition of pomegranate peel extract was determined. It was found that pomegranate peel extract shows good antifungal property. Presence of tannin is responsible for the antifungal activity of pomegranate peel extract.
Fig. 4: Zone of inhibition of pomegranate peel extract
The zone of inhibition of different batches of nail lacquer F1, F2, F3, F4, F5 was determined. The F1 and F2 batches do not show any zone of inhibition. The F3 and F4 batches show a small zone of inhibition. The F5 batch shown wide zone of inhibition therefore it is having good antifungal property compared to others.
Fig. 5: Zone of inhibition obtained from Batches F1-F5 containing pomegranate extract
In-vitro ungual permeation study of different batches of nail lacquer was performed. The zone of inhibition obtained from different batches of nail lacquer shows that the nail lacquer penetrates into the phosphate buffer solution in receptor compartment and it was responsible for showing zone of inhibition.
Fig. 6: Soaking of nail in phosphate buffer solution
Fig. 7: Zone of inhibition obtained from the sample taken from receptor compartment at 30 minutes for batch F1-F5
Fig.8: Residual film of nail lacquer after permeation study of batch F5
CONCLUSION:
The objective of the present work was to formulate a medical antifungal nail lacquer containing the extract of pomegranate peel as an antifungal agent for the treatment of onychomycosis a fungal infection affecting nail unit.
Initially the study was started with a wide thorough literature survey followed by nail lacquer containing the extract of pomegranate peel and a permeation enhancer (salicylic acid) in different concentration was tried out and comparison of extend of drug permeation has been done among the prapared5 batches (F1, F2, F3, F4, and F5). In these preparations we used nitrocellulose and ethyl cellulose as a rate controlling polymer to sustain the drug release as well as to be a good film forming agent.
Numerous investigations on physicochemical factors including film formation, salicylic acid concentration optimization, non-volatile content, drying rate, gloss, viscosity, smoothness and anti-microbial tests were conducted. And also during the formulation it was concluded that the extract used and the excipients in the formulation were compatible with each other.
All formulations showed good film formation, drying time, smooth flow and required volatile content and the results obtained from the study indicate formulation F5 showed a complete drug release, permeation rate, better smoothness and optimum drying time.
The F5 formulation had salicylic acid concentration of 9.35%w/v as keratinolytic agent and the percentage non-volatile content was found to be 17.41. Thus, with the complete evaporation of volatile content the necessary amount of non-volatile matter was observed. Also, F5 formulation demonstrated quick drying rate. Further, the formulation has a sufficient adhesive strength on the applied nail surface when compared to marketed sample. Therefore, formulation F5 was selected as the optimized nail lacquer formulation based on different evaluation studies.
Based on the findings, medicated nail lacquers showed to be a better tool as a drug delivery method for the ungula medication delivery of an anti-fungal in the treatment of onychomycosis. In addition to treating the nail infections, the medicated nail lacquer can also be used for nail beautifying with ease of application. This increases patient acceptance and compliance irrespective of the sex of patient.
CONFLICT OF INTEREST:
The authors have no conflicts of interest regarding this investigation.
ACKNOWLEDGEMENT:
We express our heartful thanks to the Management, prinCipal, Guide, Co-guide and Lab Assistants for providing all the necessary services and also for immeasurable support and encouragement for the successful completion of our work.
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Received on 01.02.2024 Revised on 11.01.2025 Accepted on 13.05.2025 Published on 30.10.2025 Available online from November 08, 2025 Research J. Topical and Cosmetic Sci. 2025; 16(2):69-76. DOI: 10.52711/2321-5844.2025.00011 ©A and V Publications All right reserved
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