INTRODUCTION:

Fungal nail infection is a common problem affecting around three in every 100 people around the world. Around half of all nail problems are due to a fungal infection. Fungal nail infection is about four times more common in toe nails than finger nails and can involve all or part of the nail, including the nail plate, nail bed and root of the nail. It usually affects adults, especially men and becomes more common as you get older. Healthy-looking nails are an important part of an individual's body image and any nail abnormalities may be considered by patients as a significant cosmetic problem, markedly influencing their self esteem¹.

However, recent data have indicated that nail lesions are not only important because of cosmetic disfigurement, but may be a symptom of significant morbidity.

Based on literature data and our own experience we conclude that nail diseases cause a marked decrease in health-related quality of life in a substantial percentage of patients. Nail changes are an important medical concern for patients and, therefore, nail diseases should raise attention and receive proper care from both physicians and other healthcare providers².

The body normally hosts a variety of microorganisms, including bacteria and fungi. Fungi can live on the dead tissues of the hairs, nails. Continuous exposal of nail to warm, moist environments usually develops nail infection. Nail plate is main route for penetration of drug³.

The Ungual drug delivery system is a new drug delivery system in the treatment of nail infections. Penetration of topical antifungal through the nail plate requires a vehicle that is specifically formulated for transungual delivery. Recent focus is emphasizing on development of a promising antifungal treatment in the form of nail lacquer owing to its beneficial advantages⁴.

Nail Disorders:

Onycholysis:

The nail becomes detached from its bed at the base and side, creating a space under the nail that accumulates dirt. Air under the nail may cause a grey-white colour but can vary from yellow to brown. If Pseudomonas aeruginosa grows underneath the nail, then green colour appears.

Paronychia:

Paronychia is inflammation of the tissue around the fingernail, with pus accumulating between the cuticle and the nail matrix. The area may become swollen, red and tender. Acute paronychia is usually due to bacterial infection, particularly Staphylococcus aureus. Chronic paronychia may be associated with eczema or psoriasis. It is often due to Candida infection but other pathogens - eg, Pseudomonas species. (Producing a green or black discoloration) - may be the cause.

Psoriasis:

Virtually all patients with psoriasis have nail involvement at some time and it occurs in 50% of cases at any given time.

Yellow nail syndrome:

Yellow nail syndrome is characterized by slow-growing, excessively curved and thickened yellow nails which are associated with peripheral lymphoedema and exudative pleural effusions. Abnormalities include nail pits, transverse furrows, crumbling nail plate and roughened nail.

Amorolfine nail lacquer contains the active substance amorolfine (as amorolfine hydrochloride), which belongs to a group of medicines known as antifungal. It kills a wide variety of fungi that can cause nail infections. Amorolfine nail lacquer is used to treat fungal infections of the nails. Amorolfine 5% w/v medicated nail lacquer contains 5.574gms of amorolfine hydrochloride per 100 ml equivalent to 5 gm (5%w/v) of amorolfine base. Its fungi static and fungicidal efficacy is based on the alteration of fungal cell membrane targeted primarily on stereo biosynthesis. The ergosterol content is reduced and at the same time unusual sterically nonpolar sterols accumulate. Amorolfine is a broad spectrum antimitotic. It is highly active against current or casual agents of onchomycosis^5,6.

Hence there is a need for the development of ungual drug delivery system containing amrolfine for fungal infection of nail.

MATERIALS AND METHODS:

Preparation of Nail Lacquers:

Amorolfine was used 5 %w/v 2.5 gm was used in 50 ml formulation; the concentration of amorolfine in all formulation was kept constant. The nail lacquer is formulated by simple mixing method. Required quantity of Eudragit RL-100 was added then weighed amount of ethyl cellulose was added, then to this mixture amorolfine, triacetin, and ethyl acetate was added and mixed until no lumps, the volume is made up to 50 ml with ethyl alcohol.

The composition of prepared formulations of nail lacquers is given in following table.

Table 1: Formulation of Nail Lacquers

Ingredients	F 1	F 2	F 3	F 4	F 5	F 6
Amorolfine	5% w/v	5% w/v	5% w/v	5% w/v	5% w/v	5% w/v
Eudragit RL 100 (gm)	0.65	0.65	1.9	1.9	2.55	2.55
Triacetin (ml)	2.5	3	2.5	3	2.5	3
Ethyl acetate (ml)	12	12	14	14	16	16
Ethyl cellulose (g)	5	6	5	6	5	6
Ethyl alcohol (ml)	q.s to 50ml	q.s to 50 ml	q.s to 50 ml	q.s to 50 ml	q.s to 50 ml	q.s to 50 ml

Evaluation of Nail Lacquers:

Non volatile content:

A sample was taken in a glass petri dish of about 8 cm in diameter. Samples were spread evenly with the help of tared wire. The dish was placed in the oven at 105ºC for 1 h then petri dish was removed, cooled and weighed. The difference in weight of sample after drying was determined^7-9.

Drying time:

A film of sample was applied on a glass petri dish with the help of brush. The time to form a dry‐to‐touch film was noted using a stopwatch¹⁰.

Smoothness:

The smoothness was determined by pouring the sample of nail lacquer approximately 1.5 inches on a glass plate and made to raise vertically¹¹.

Gloss:

Gloss of the film was visually seen, within the specifications¹¹.

Water resistance:

This is the measurement of the resistance towards water permeability of the film. A thin film on three glass plates should be applied and dried then weigh these. The plates should then be immersed in a water bath containing distilled water at 37 ⁰C for 24 h. The panels should then be removed and dried by placing the plate between absorption papers and reweighed. The increase in weight is calculated. Higher the increase in weight lowers the water resistance¹².

Viscosity:

The viscosity of nail lacquer was measured using Brookfield viscometer¹³. Samples are taken in a closed jar to minimize the solvent evaporation. At 25 ^°C, shake vigorously, start timer, insert spindle into the sample to the scored line, with the motor running at 60RPM. Read the instrument at the end of ten minutes then switch the speed control to 6 RPM and read the instrument again at the end of another ten minutes. In order to convert the dial reading to centipoises, multiply by following factor- 60rpm X 20; 6rpm X 200. The viscosity-thixotropy relationship of cream nail should be about 375-500cps at 60 rpm, at 25 °C¹⁴.

Colour:

Colour comparing with master color standards by applying on thumbnails, holding them side by side, moving the thumb with the standard first on the right and then on left. Artificial acrylic nails have been utilized as well for matching comparative shades¹⁵.

Stability Studies:

According to ICH guidelines at 40 ± 2 ^oC/75 ± 5% RH sample was stored in stability chamber for one month. The sample was evaluated for non volatile content, drying time, gloss, smoothness of flow, water resistance.

Antifungal activity of nail lacquers by disc diffusion method:

The antifungal activity was screened by disc diffusion method¹⁶. The Sabouraud’s dextrose agar plates were inoculated with the fungal culture of Candida albicans (72h culture). Sabouraud Dextrose Broth cultures were prepared in test tubes. Using sterile cotton swab, the surface of Sabourauds Dextrose Agar plates were swabbed to prepare lawn cultures. 5 min after the agar surface had dried, wells were dug using sterile cork borer aseptically. Wells were saturated with nail lacquers six formulations. Standard fluconazole disc was used for comparison. The plates were incubated at 28 ºC for 24-48 hrs. The diameters of zones of inhibition were measured using a scale to the nearest millimeter.

RESULTS AND DISCUSSION:

Six formulations were prepared and evaluated for the physical properties.

Physical evaluation:

Table 2: Evaluation of Non volatile content, Drying Time and Smoothness

Formula	% Non volatile content ±SD	Drying Time (sec )±SD	Smoothness
F1	25% ± 0.23	63 ± 0.25	++
F2	28% ± 0.24	65 ± 0.23	++
F3	31%±0.28	68 ± 0.29	++
F4	33%±0.24	69 ± 0.26	++
F5	35%±0.26	70 ± 0.24	+++
F6	36%±0.25	75 ± 0.29	+++

+= Satisfactory, ++ = good, +++ = very good

The data of non-volatile content, drying time and smoothness is given in the above table 2. Desired amount of non-volatile matter was seen with complete evaporation of volatile-matter leaving a thin film. It ranges from 25-36%. It was seen that as the polymer concentration increased non-volatile content also increased and vice-versa. Non-volatile content depends and vary upon the concentration of polymer used.

The application and performance properties of nail coating depend greatly upon volatility characteristics of its solvent system and therefore on its drying time. The values of the drying time shown in the above table 2. Drying time of nail lacquer was found in the range of 63-75%. As the polymer concentration increased drying time increased.

This is a character of a film. Formulation F1-F4 showed good smoothness due to lower polymer concentration and formulation F5-F6 shows very good smoothness due to presence of increased polymer concentration.

Table 3: Evaluation of Water Resistance, Viscosity, Colour and Gloss.

Formulation	Water Resistance (gm)	Viscosity (cps) ±SD	Colour	Gloss
F1	0.03	242.6 ± 0.24	Transparent	+++
F2	0.04	263.4 ± 0.19	Transparent	+++
F3	0.06	298.5 ± 0.23	Transperent	+++
F4	0.07	302.7 ± 0.25	Transperent	+++
F5	0.10	312. 3 ± 0.27	Transparent	++
F6	0.11	319.4 ± 0.20	Transparent	++

+ = Satisfactory, ++ = good, +++ = very good

Results of water resistance test are given in the above Table 3. It can be seen that as the polymer concentration increased water resistance also increased and as the polymer concentration decreased water resistance also decreased. Lower the increase in the weight of nail lacquer film higher is the water resistance capacity. F1, F2, F3, F4 showed lower water resistance compared to F5 and F6.

Viscosity measurements are done as per the procedure mentioned in the methodology and results are given in the above Table 3. Viscosity is an important parameter for characterizing the nail lacquer as it effects the spreadablity and release of drug. All formulation showed increased viscosity ranging from 240-320 cps. As the polymer concentration increased viscosity also increased and vice-versa.

All the formulation prepared it was observed that the color was found to be transparent. And there is no significance change in the color.

The results of the gloss are shown in the Table 3. The character of the film was visually seen. Formulation F1, F2, F3, F4 showed good gloss compared to F5 and F6. The gloss of the formulation with more polymer concentration was less compared to formulation with less polymer concentration.

Antifungal activity of nail lacquers by disc diffusion method

In anti fungal activity studies, among all the formulations F5 and F6 formulation showed better anti fungal activity as compared to fluconazole. The results are shown in following table 4

Stability Studies:

The stability test is carried out for 8 week and there is no significant change is observed in color, non volatile content, viscosity, drying time and smoothness.

CONCLUSION:

The present work was to develop and evaluate ungula drug delivery system containing amorolfine using different polymers. This drug delivery system was found to have good antifungal property. This nail lacquer was found to have no side effects. The prepared nail lacquers were evaluated for physical characteristics which showed good results within standard specifications. Stability studies showed that there were no variations in the evaluation parameters in the prepared formulations. Anti fungal activity of all the formulations of nail lacquers showed significant zones of inhibition compared with standard antifungal agent Fluconazole. Thus it can be concluded that use of ungual drug delivery system with Amorolfine is capable of treating fungal infections of nail.

REFERENCES:

1. Baran R. Amorolfine nail lacquer. A new Transungual Delivery System for nail mycoses. J Am Med Assoc Southeast Asia. 9(4); 1993: 5–6.

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11. Mithal BM, Saha RN. A handbook of cosmetics, 1st edition reprint 2005, Vallabh prakashan, Delhi, 176- 182.

12. Elezovic A, Hadzovic S, Hadzidedics S, Kostic S. Characterization of antifungal nail lacquer formulation containing fluconazole. Scientia Pharmaceutica. 78; 2010: 624-628.

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14. Khar RK, Nanda A, Nanda S. Cosmetic technology, 1st edition 2002, Birla publications, Delhi, 12- 22.

15. Indian standard of Nail Polish (Nail enemal) specification (IS 9542:1994). Bureau of Indian standard, 2003, pp: 35

16. Kalemba D Kunicka A. Antibacterial and antifungal properties of essential oils. Cur

Received on 23.12.2016 Modified on 07.01.2017

Research J. Pharm. and Tech. 2017; 10(3): 869-872.

DOI: 10.5958/0974-360X.2017.00162.7

Organism: candida albicans	F1	F2	F3	F4	F5	F6	Fluconazole
Zone of inhibition in mm	31	34	32	35	39	40	42