INTRODUCTION:

In recent years, the number of Asian women aspiring for a whiter skin complexion has increased dramatically. This is due partly to the discovery of many effective skin whitening agents, particularly those derived from plants. The skin colour is mainly determined by the content of an epidermal pigment called melanin. It is secreted by melanocyte cells in the basal layer of the epidermis. Melanin may be overproduced due to chronic sun exposure, melasma, reactive oxygen species (free radicals) or other hyperpigmentation diseases. The excessive melanin production is not desirable since it may cause a darker or uneven skin colour. Changes in colour are also desired for cosmetic reasons. The initial process of melanin production (melanogenesis) is controlled by tyrosinase, which is an enzyme catalysing the hydroxylation of tyrosine, the precursor of melanin, into dihydroxyphenylalanine and other intermediates. Thus, inhibition of tyrosinase activity or its production can prevent melanogenesis, and reducing the uneven skin colour. Hence the plants produces strong anti-tyrosinase activity is used for skin whitening agent in cosmetic preparations^1-3.

The herbal products claim to have no side effects, commonly seen with products containing synthetic agents. Attractiveness of herbal preparations has socially as well as technologically resulted in flooding of market place in India⁴. So here we planned to select such type of plants which are found to exhibit anti-tyrosinase activity for preparation of polyherbal cream. The plants used in cosmetic preparation have varieties of property like anti-tyrosinase, antioxidant, anti-inflammatory, antiseptic and antibacterial etc. As per this concern we select following plants Curcuma longa, Azadirachta indica, Coriandrum sativum, Trigonella foenum-graeceum, Mentha arvensis, Cocos nucifera and Prunus amygdalus oil for the formulation of herbal cosmetic.

Curcuma longa rhizomes are valued as a topical antioxidant and anti-inflammatory agent, with superior free radical scavenging and lipid peroxidation inhibition efficacy as compared to vitamin E. The curcumin and Tetrahydrocurcuminoids present in C. longa efficiently inhibits the tyrosinase. The parent compound Curcumin is a potent inhibitor of protein kinase C, EGF-receptor tyrosine kinase and IkappaB kinas. Moreover C. longa is reported an effective skin lightening agent with multifunctional topical benefits, without irritant and sensitization side effects⁵. The saponin glycosides are present in Trigonella foenum-graeceum seeds and is claimed to be useful for prevention of inflammation and melanin synthesis that occur during skin damage or as a result of dermatological diseases⁶. The Coriandrum sativum also produce anti-tyrosinase⁷ activity and Prunus amygdalus is enriched with Vitamin E⁸. The Azadirachta indica⁹ are reported to possess highly antibacterial property, hence it is required to inhibit the microbial growth when skin become humectants. Mentha arvensis were used as cooling and soothing agent¹⁰, while Cocos nucifera oil is useful for skin itching and rashes¹¹. In the present study, an attempt has been made to combine all these plants in preparation of polyherbal cosmetic cream and to produce the synergistic whitening effects on face skin.

MATERIAL AND METHODS:

Preparation of extracts

Air dried and coarsely powdered (500 gm) of Curcuma longa rhizomes, Azadirachta indica leaves, Coriandrum sativum seed, Trigonella foenum-graeceum seeds and Mentha arvensis leaves were placed in soxhlet extractor separately, using petroleum ether and then successively with ethanol. The extracts were then concentrated to dryness under reduced pressure and controlled temperature, respectively and they were preserved in a refrigerator.

The Cocos nucifera and almond oil were collected from local market of Bhopal (M.P.).

Preparation of cream base:

Oil in water (O/W) emulsion-based cream (semisolid formulation) was formulated. The emulsifier (stearic acid) and other oil soluble components (Cetyl alcohol, almond oil) were dissolved in the oil phase (Part A) and heated to 75° C. The preservatives and other water soluble components (Methyl paraban, Propyl paraban, Triethanolamine, Propylene glycol) 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 until cooling of emulsifier took place. The formula for the base is given in table 1.

Table 1: Composition of cream base

Ingredients	Formula % w/w
Ingredients	F1	F2	F3	F4	F5	F6
Stearic acid	15	12	12	10	18	15
Cetyl alcohol	2	3	4	4	3	4
Almond oil	4	4	4	4	4	4
Glycerol	3	3	3	3	3	3
Methyl paraban	0.02	0.02	0.02	0.02	0.02	0.02
Triethanolamine	qs	qs	qs	qs	qs	qs
Water, qs, 100	qs	qs	qs	qs	qs	qs

Table 2: pH of cream base

Formulation	pH
F1	6.5
F2	6.7
F3	6.8
F4	6.8
F5	6.4
F6	6.5

Drug formulation:

The appropriate base was selected from table and cream was formulated. The emulsifier (stearic acid) and other oil soluble components (Cetyl alcohol, almond oil) were dissolved in the oil phase (Part A) and heated to 75° C. The preservatives and other water soluble components (Methyl paraban, Propyl paraban, Triethanolamine, Propylene glycol, all extracts) 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 until cooling of emulsifier took place. The composition of cream is given in table 6.

Evaluation of cream

pH of the Cream:

The pH meter was calibrated using standard buffer solution. About 0.5g 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 100 rpm, using spindle no 7.

Dye test:

The scarlet red dye is mixed with the cream. Place a drop of the cream on a microscopic slide covers it with a cover slip, and examines it under a microscope. If the disperse globules appear red the ground colourless. The cream is o/w type. The reverse condition occurs in w/o type cream i.e. the disperse globules appear colourless in the red ground.

Homogeneity:

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

Appearance:

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

After feel:

Emolliency, slipperiness and amount of residue left after the application of fixed amount of cream was checked.

Type of smear:

After application of cream, the type of film or smear formed on the skin were checked.

Removal:

The ease of removal of the cream applied was examined by washing the applied part with tap water.

Acid value:

Take 10 gm of substance dissolved in accurately weighed, in 50 ml mixture of equal volume of alcohol and solvent ether, the flask was connected to reflux condenser and slowly heated, until sample was dissolved completely, to this 1 ml of phenolphthalein added and titrated with 0.1N NaOH, until faintly pink color appears after shaking for 30 seconds.

Acid value = n*5.61/w

n = the number of ml of NaOH required.

w = the weight of substance.

Saponification value:

Introduce about 2 gm of substance refluxed with 25 ml of 0.5 N alcoholic KOH for 30 minutes, to this 1 ml of phenolphthalein added and titrated immediately, with 0.5 N HCL.

Saponification value = (b-a)*28.05/w

The volume in ml of titrant = a

The volume in ml of titrand =b

The weight of substance in gm = w

Irritancy test:

Mark an area (1sq.cm) on the left hand dorsal surface. The cream was applied to the specified area and time was noted. Irritancy, erythema, edema, was checked if any for regular intervals up to 24 hrs and reported.

Accelerated stability testing:

Accelerated stability testing of prepared formulations was conducted for 2 most stable formulations at room temperature, studied for 7 days. They were formulation number 4 and 5 at 40 ^oC ± 1 ^oC for 20 days. The formulations were kept both at room and elevated temperature and observed on 0th, 5th, 10th, 15th and 20th day for the following parameters^12-18.

RESULTS:

pH of the Cream:

The pH of the cream base was found to be in range of 6-7 which is good for skin pH. All the formulations of cream base were shown pH nearer to skin required (Table 2).

Viscosity:

The viscosity of was cream was in the range of 27020-27054 cps which indicates spreadibilty of cream. In our study F₂, F₃ and F₄ depicted easily spreadable by small amounts of shear, while F₁, F₅ and F₆were not easily spreadable on skin. But F3 shows good spreadable property than other formulations.

Acid value and Saponification value:

The results of acid value and saponification value of all formulation of cream base were presented in table 3, and showed satisfactorily values.

Irritancy test:

The formulation F3 shows no redness, edema, Inflammation and irritation during irritancy studies. These formulations are safe to use for skin (Table 4).

Dye test:

This dye confirms that all formulations were o/w type emulsion cream. But formulation (F3) shows more stable in o/w type emulsion. So here we select F3cream base for further study.

Table 3: Test applied for acid value and saponification value

Parameter	Formula
Parameter	F1	F2	F3	F4	F5	F6
Acid value	6.5	5.4	6.3	5.9	5.1	6.1
Saponification value	25.4	27.2	26.7	26.4	25.3	24.7

Table 4: Type of adverse effect of formulations

Formulation	Irritant	Erythema	Edema
F1	NIL	NIL	NIL
F2	NIL	NIL	NIL
F3	NIL	NIL	NIL
F4	NIL	NIL	NIL
F5	NIL	NIL	NIL
F6	NIL	NIL	NIL

Homogeneity:

All formulations produce uniform distribution of extracts in cream. This was confirmed by visual appearance and by touch (Table 5).

Appearance:

When formulation were kept for long time, it found that no change in colour of cream (Table 5).

After feel:

Emolliency, slipperiness and amount of residue left after the application of fixed amount of cream was found (Table 5).

Type of smear:

After application of cream, the type of smear formed on the skin were non greasy (Table 5).

Removal:

The cream applied on skin was easily removed by washing with tap water (Table 5).

Cream:

From above study, the F3 base was selected for the preparation of herbal cream, and the composition of cream has illustrated on table 6. The physical evaluation and stability of polyherbal cream has shown in table 7, and results were considerable and acceptable.

DISCUSSION:

Melanin is the primary determinant of human skin colour. Regular use of herbal face cream can reduce the production of melanin. The principle constituents present in C. longa, C. sativum and T. foenum-graeceum have potent anti-tyrosinase and antioxidant activity, and play synergistic action on reduction of melanin production. The antibacterial property of A. indica produces protection from bacterial infection. The vitamin E present in P. amygdalus oil and its antioxidant property protect the skin from premature ageing. For improvement of herbal cream of whitening effect in skin should be well hydrated and protected from environmental damage. Here we assure that the herbal face cream can produce excellent whitening effect on skin due to synergistic anti-tyrosinase, antioxidant, anti-inflammatory and antibacterial properties of the ingredients.

Table 5: Physical parameter of F3 cream base on room and accelerated temperature

Days	Temperature	Parameter
Days	Temperature	pH	X₁	X₂	X₃	X₄	X₅	X₆
0	RT	6.9	**	NCC	**	E	NG	ES
0	40 ºC + 1 ºC	6.8	**	NCC	**	E	NG	ES
5	RT	6.7	**	NCC	**	E	NG	ES
5	40 ºC + 1 ºC	6.8	**	NCC	**	E	NG	ES
10	RT	6.6	**	NCC	**	E	NG	ES
10	40 ºC + 1 ºC	6.7	**	NCC	**	E	NG	ES
15	RT	6.7	**	NCC	**	E	NG	ES
15	40 ºC + 1 ºC	6.8	**	NCC	**	E	NG	ES
20	RT	6.7	**	NCC	**	E	NG	ES
20	40 ºC + 1 ºC	6.7	**	NCC	**	E	NG	ES

X₁-Homogenity, X₂-Appearance, X₃-Spreadibility, X₄-After feel, X₅-Type of smear, X₆-Removal, **: Good, *: Satisfactory, E: Emollient, NG: Non greasy, ES: Easy, NCC: Not change in colour

Table 6: Composition of polyherbal cream

Ingredients	Formulation (% w/w)
Ethanol extract of A. indica	0.75
Ethanol extract of T. foenum	0.20
Ethanol extract of C. longa	0.30
Ethanol extract of C. sativum	0.25
Ethanol extract of M. arvensis	0.30
Coconut oil	0.20
Stearic acid	12
Cetyl alcohol	4
Almond oil	4
Glycerol	3
Methyl paraban	0.02
Triethanolamine	qs
Water, qs, 100	qs

Table 7: Evaluation of herbal cream

Formulation	pH	Acid value	Saponification value	Adverse effect	Physical Parameter
					Room Temperature	Accelerated Temperature
Herbal cream	6.9	6.3	26.3	NIL	Acceptable	Acceptable

Our study indicated that the base F3 found to be more stable, while remaining base were not stable and resulted in breakdown of the emulsion when stored for long time. So that base F3 was appropriate for development of polyherbal cream, hence we prepared polyherbal cream by mixing all the extract in this base. The pH of prepared cream was nearer skin pH, and cream produces homogeneous, emollient, non-greasy and easily removed properties after the application. The polyherbal cream was safe in respect to skin irritation and allergic sensitization. The finding report illustrated that polyherbal face cream is effective in whitening the skin complexion, and making it soft and smooth. The study suggests that herbal face cream is more stable, safe and efficacious.

ACKNOWLEDGEMENTS:

The authors acknowledge to Director of Oriental College of Pharmacy, Bhopal (M.P.), India, for providing all the facilities and successfully completion of work.

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Received on 17.06.2012 Accepted on 30.06.2012

Res. J. Topical and Cosmetic Sci. 3(1): Jan. –June 2012 page 23-27