Formulation and Evaluation of a Poly Herbal Skin Care Cream containing Neem and Tulsi

 

R. Chandrasekar*, B. Sivagami

Department of Pharmacognosy, Seven Hills College of Pharmacy, Tirupati.

*Corresponding Author E-mail: chandrumnrcop@gmail.com

 

ABSTRACT:

Neem and Tulsi called as holy tree and herb has been used since ancient times for the treatment of various skin diseases. In this study Neem leaves Azadirachtaindica and Tulsi leaves Ocimum sanctum was incorporated with various ingredients and a poly herbal cream was prepared. Neem leaves and Tulsi leaves were collected from Tirupati Local Area. Both these plants were shade dried for 4 days and size reduced using mixer grinder converted into coarse powder and passed through sieve number 22. The coarse powder was stored for further studies. Both Neem and Tulsi leaf were defatted using pet ether 60-40 and then extracted using ethanol in a soxhlet apparatus. The extraction was carried out for 3 hrs taking 100 gms of plant material with 500 ml of ethanol. The extract was evaporated in a rotary evaporator the obtained ethanol extracts were preserved for further studies in a dessicator. The extracts were incorporated with various ingredients and excipients and a poly herbal cream was formulated. The cream was evaluated for various evaluation parameters which include pH, viscosity, spreadability, centrifugation test, spectrophotometric test, accelerated stability studies and microbial stability. The accelerated stability studies were carried out for 20 days in room and elevated temperature in a stability chamber. The cream was found to be stable after centrifugation there was no phase separation after centrifugation and stability study. The microbial stability was evaluated for bacterial growth (E. coli) and yeast. The results revealed that the microbial stability of the cream incorporated no microbial growth.

 

KEYWORDS: Azadirachtaindica, Ocimum sanctum, Evaluation,Poly Herbal, Stability.

 


 

 

INTRODUCTION:

Neem is a tree Azadirachtaindica belonging to Meliaceae Family. The bark, leaves, and seeds are used to make medicine. Less frequently, the root, flower, and fruit are also used. Neem leaf is used for leprosy, eye disorders, bloody nose, intestinal worms, stomach upset, loss of appetite, skin ulcers, diseases of the heart and blood vessels (cardiovascular disease), fever, diabetes, gum disease (gingivitis), and liver problems. The leaf is also used for birth control and to cause abortions. Some people apply neem directly to the skin to treat head lice, skin diseases, wounds, and skin ulcers; as a mosquito repellent; and as a skin softener. Neem is also used as an insecticide. [1]

Of all the herbs used within Ayurveda, tulsi (Ocimum sanctum Linn) is preeminent, and scientific research is now confirming its beneficial effects. There is mounting evidence that tulsi can address physical, chemical, metabolic and psychological stress through a unique combination of pharmacological actions. Tulsi has been found to protect organs and tissues against chemical stress from industrial pollutants and heavy metals, and physical stress from prolonged physical exertion, ischemia, physical restraint and exposure to cold and excessive noise. Tulsi has also been shown to counter metabolic stress through normalization of blood glucose, blood pressure and lipid levels, and psychological stress through positive effects on memory and cognitive function and through its anxiolytic and anti-depressant properties. Tulsi's broad-spectrum antimicrobial activity, which includes activity against a range of human and animal pathogens, suggests it can be used as a hand sanitizer, mouthwash and water purifier as well as in animal rearing, wound healing, the preservation of food stuffs and herbal raw materials and traveler's health. Cultivation of tulsi plants has both spiritual and practical significance that connects the grower to the creative powers of nature, and organic cultivation offers solutions for food security, rural poverty, hunger, environmental degradation and climate change. The use of tulsi in daily rituals is a testament to Ayurvedic wisdom and provides an example of ancient knowledge offering solutions to modern problems. [2]

 

Several medicinal plants have been used as cosmetics since time immemorial, and showed promising effects on various skin infections like acne, blackheads, age spots, skin rashes, skin allergy, ageing of skin, wrinkles, skin whitening agents etc. Some very common plants like Aloe vera ,Azadirachtaindica , Annona squamosa, Aterocarpusheterophyllus, Carica papaya , Centellaasiatica, Curcuma longa, Mangiferaindica, Moringa citrifolia, Ocimum sanctum , Phyllanthus emblica , Psidium gujava, Terminalia arjuna, Terminalia chebula, Vitis vinifera etc. have been extensively reported in Ayurveda, Siddha and Unani systems of medicines for their cosmetic potential. [3]

 

Boswellic acid, tetrahydrocurcuminoids, retenoids, panthenol, glucopyranosides, hydroxy acids, antioxidants, vitamin E etc. these are some active constituents present in herbs which act on the skin and used as cleansing agent, toners and moisturizers. They help the skin in many ways in enhancing the beauty and protecting the skin from infections. The objective of present study is to prepare skin care product that not only moisturizes and softens the skin but also helps in healing of skin diseases and skin infections. An herbal cream that can give effective protection to skin and free from any side effects or toxic residue or any irritation when regularly used and should also be cosmetically acceptable. [4, 5]

 

MATERIALS AND METHODS:

Collection of plant material:

Neem Azadirachtaindica leaves and Tulsi leaves Ocimum sanctum were collected from local area from Tirupati. The plants were identified by Prof. N. Yasodamma, S.V. University, Tirupati. Specimen no: K.H-01 (Azadirachtaindica) and Specimen no: K.S.-01 (Ocimum sanctum)

 

Chemicals and Reagents

Almond oil was obtained from Dabur India limited, Mentha oil was obtained from Himalaya herbals Pvt Ltd. Stearic acid, Lecithin, Poly Ethylene Glycol, Sodium Laurel Sulphate, Triethanol amine, were obtained from Sigma Aldrich Mumbai. All the other chemicals were obtained from SD Fine Chem Pvt. Ltd.

 

Instruments and Equipments

Double Beam UV Spectrophotometer-Analytical technologies Limited, Model 212R RI, Centrifuge-R-8C REMI Instruments, Digital pH meter Systronics, Mumbai, Brookfield Viscometer Servewell Pvt. Ltd. Model Number. LVDVE, SUPERFIT ROTAVAP, Model-PBU-6. Servewell Instruments Pvt. Ltd.

 

Extraction of plant material

Both Neem and Tulsi leaves were shade dried for 4 days and size reduced using mixer grinder converted into coarse powder and passes through sieve number 22. The coarse powder was stored for further studies. Both Neem and Tulsi leaves were defatted using pet ether 60-40 and then extracted using ethanol in a soxhlet apparatus. The extraction was carried out for 3 hrs taking 100 gms of plant material with 500 ml of ethanol. The extract was evaporated in a rotary evaporator (SUPERFIT ROTAVAP, Model-PBU-6) the obtained ethanol extracts were preserved for further studies in a dessicator.

 

Preparation and formulation of poly-herbal cream

The poly-herbal cream containing Azadirachtaindica leaf ethanol extract NLE and Ocimum sanctum leaf ethanol extract TLE were formulated. Oil in water O/W cream was formulated using, the oily phase (Part A) and the aqueous phase (Part B). Oil in water (O / W) emulsion - based cream (semisolid formulation) was formulated. The emulsifier (stearic acid) and other oil soluble components (Lecithin, Almond oil and fraction of ethanolic extract of Azadirachtaindica leaf and Ocimum sanctum leaf ethanol extract were dissolved in the oil phase (Part A) and heated to 75 C. The preservatives and other water soluble components, (PEG, SLS, triethanolamine, propyl parabene, Mentha oil and water) 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. [6, 7] the formula for the cream is given in Table 1.

 


Table 1. Formula for cream

Part A (Oil Phase)

Part B (Aqueous Phase)

Ingredients % w/ w

C1% w/ w

C2 % w/ w

Ingredients % w/ w

C1% w/ w

C2 % w/ w

Stearic acid

(Emulsifier)

--

3%

Poly ethylene glycol

(Base)

2%

5%

Lecithin

(Surfactant)

3%

--

Sodium laurel sulphate

(Surfactant)

3%

3%

Almond oil

(Emollient)

4%

4%

Triethanol amine

 

1%

2%

NLEE

 

2%

5%

Propyl Parabene

(Preservative)

0.02%

0.18%

TLEE

 

2%

5%

Mentha oil

(Perfume)

0.02%

0.02%

 

 

 

Water upto 100%

QS upto 100%

QS upto 100%

 

The herbal formulation was prepared using cream base incorporating all necessary ingredients along with the extracts of Azadirachtaindica and Ocimum sanctum. Formulation was then evaluated for its physical properties.

 

 

 

Figure 1. Cream 1

Figure 2 .Cream 2


Evaluation of the cream

The cream was evaluated for its physiochemical parameters pH, Viscosity, Dye test, Homogeneity, Appearance, After feel, Type of smear, Removal, Acid value, Irritancy test, Accelerated stability testing. [8, 9]

 

pH of the Cream

The pH of the cream was determined using a pH meter and 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. Digital pH meter Systronics

 

Viscosity

Viscosity of the formulation was determined by using Brookfield Viscometer at different RPMs, spindle number-64 was used for the study. Brookfield Viscometer Servewell Pvt. Ltd. Model Number. LVDVE

 

Figure 3 Brook Field Viscometer

Dye test

Ruthenium red dye was used for dye test. A drop of the cream was placed on a microscopic slide covered with a cover slip, and examined under a microscope. If the disperse globules appear red the ground colorless. The cream is o/w type. The reverse condition occurs in w/o type cream i.e. the disperse globules appear colorless in the red ground.

 

Homogeneity

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

 

Appearance

The appearance of the cream was studied by its color, pearl scence 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.

 

Spreadability

Spreadability denotes the extent of area to which the formulation readily spreads on application to skin or hair. The bioavailability efficiency of a formulation also depends on its spreading value. [10-12]Spreadability is expressed as

 

S = m * l/t

m weight tied on upper slide

l length of glass slide

t time in s

 

Spectrophotometric test

The cosmetic formulations were diluted in ultra-pure water at ratio of 1/100 (w/v) and then submitted to scanning analysis by spectrophotometry in the UV-VIS region (245 nm). (Double Beam UV Spectrophotometer-Analytical technologies Limited, Model 212R RI).

 

Centrifugation test

To perform the centrifugation test, 5 g of sample was subjected to a cycle of 3000 rpm for 30 minutes at room temperature. At the end of the centrifugation period, the cosmetic formulations were examined for phase separation which is an indication of cosmetic formulation instability. (Centrifuge-R-8C REMI Instruments).

 

Accelerated stability testing

Stability testing was carried out for Formula no. 2 by keeping 50 g of cream at 40C1C and another 50 g at room temperature. It was checked for any visual appearance and phase separation from time to time over a period for 20 days.

 

In order to determine the viscosity the cream was kept at room temperature and at an elevated temperature of 40C1C. For this, 50 g of the cream base was kept at 40C1C in stability chamber. Stability of this cream was measured after regular intervals of time for 20 days. [10- 12]

 

RESULTS AND DISCUSSION:

pH of the Cream:

The pH of the cream was found to be in range of 5.9 to 6.9 which is good for skin pH. The cream has shown pH nearer to skin required i.e pH of C2 was found to be 6.9.

 

 

Figure No 4 pH of the cream

 

Viscosity:

The viscosity of cream was in the range of 50910 40900 cps which indicates that the cream is easily spreadable by small amounts of shear. But the C2 shows good spreadable property than the other formulation. The results are given in Table no 5 and Fig no 5

 

Spreadability Test

Spreadability denotes the extent of area to which the formulation readily spreads on application to skin or hair. The bioavailability efficiency of a formulation also depends on its spreading value.

 

Table No 2 Spreadability Index

S. NO

Formulation 1

Formulation 2

1

14.9

9.9

2

7.5

8

 

Figure No 5 Spreadibility Index

 

Acid value and Saponification value:

The results of acid value and saponification value of the formulation of cream are presented in Table 3, and showed satisfactorily values. [8, 9]

 

Spectrophotometric test:

The cosmetic formulations were diluted in ultra-pure water at ratio of 1/100 (w/v) and then submitted to scanning analysis by spectrophotometry in the UV-VIS region (245 nm).

 

Dye test:

This dye confirms that the formulation was o/w type emulsion cream. But formulation (C2) shows more stablility in o/w type emulsion.

 

Homogeneity:

The formulations produced uniform distribution of extracts in cream. This was confirmed by visual appearance and by touch. (Table 6)

 

Appearance:

When formulation were kept for long time, it found that no change in colour of cream. (Table 6)

 

After feel:

Theamount of residue, Emolliency of the cream and slipperiness was found. (Table 6)

 

Type of smear:

The type of smear formed may be non greasy. (Table 6)

 

Removal:

Sincethe cream C2 is O/W type it can be easily removed by washing with tap water. (Table 6)

 

Table No. 3 Viscosity of the cream

Days

C2

C1

5

50000

45000

10

40000

39000

15

39000

35000

20

40000

38000

Figure No 6 Viscosity cps

 

 

Table No. 4 Acid Value and Saponification value

S. No

Parameter

C2

1

Acid value

5.56

2

Saponification value

12.33

 

Table No. 5 Spectrophotometric test

Concentration

Absorbance

0.01g/ml

0.060

0.02g/ml

0.090

0.03g/ml

0.135

0.04g/ml

0.169

0.05g/ml

0.233

 

Figure No 7 Spectrophotometric test

 

Figure No 8 Spectrophotometric test


 

Table No. 6. Physical parameter of cream on room and accelerated temperature

Days

 

Temperature

Formulation

Parameter

pH

Homogeneity

Appearance

Spreadability

After Feel

Type of Smear

Removal

0

 

RT

40 C + 1 C

C2

6.6

Good

NCC

Good

Emollient

Non greasy

Easy

C2

6.5

Good

NCC

Good

Emollient

Non greasy

Easy

5

RT

40 C + 1 C

C2

6.4

Good

NCC

Good

Emollient

Non greasy

Easy

C2

6.3

Good

NCC

Good

Emollient

Non greasy

Easy

10

 

RT

40 C + 1 C

C2

6.4

Good

NCC

Good

Emollient

Non greasy

Easy

C2

6.5

Good

NCC

Good

Emollient

Non greasy

Easy

15

 

RT

40 C + 1 C

C2

6.5

Good

NCC

Good

Emollient

Non greasy

Easy

C2

6.3

Good

NCC

Good

Emollient

Non greasy

Easy

20

RT

40 C + 1 C

C2

6.6

Good

NCC

Good

Emollient

Non greasy

Easy

C2

6.3

Good

NCC

Good

Emollient

Non greasy

Easy

NCC: Not change in colour

 

 

 


Microbial enumeration test and absence of specified microorganism

The microbial stability of the cosmetic formulations was evaluated through the microbial contamination test. After being prepared the culture media were autoclaved at 125C for 20 minutes and then 20 mL of the culture medium was poured into a sterile petri dish. Then 0.2g of the formulation was placed in the center of each petri dish, and then the plates were incubated at 37C or at 25C for 3 days according to the inoculated microorganisms. After the incubation period, plates were taken out and checked for microbial growth, which is an indication of contamination. Sample dissolved in phosphate buffer pH 7.2, shaken well and made upto 100ml. pipette out 1 ml of dilution into a sterile petridish, pour 30ml of the soyabean casein digest agar, and mix well. Solidify the agar and invert and incubate at 30o C to 35o C for 48 to 72 hrs for Bacterial count. To the specimen of the sample add fluid lactose medium and make up to 100 ml, incubated for 24 hrs at 37 o C for E. coli. Incubate at 30o to 35o for 48 to 72 hrs for presence of bacteria. Similarly pipette out 1 ml of dilution into a sterile petridish and pour 30 ml of the sabour and dextrose agar mix solidify invert and incubated at 20 o C to 25 o C for 5 days for yeast. The contamination test was performed using a variety of microorganisms namely Bacteria (Escherichia coli) and Yeast. After the incubation period, the plates were taken out and checked for microbial growth by comparing it with the control. No microbial growth was observed in the formulation. Figures 7-14. The obtained results confirmed that there was no contamination in the formulation. But formulation C1 was found to be contaminated formulation C1 was discontinued from the study. [13]

 

 

 

 

 

 

Formulation C2

 

Figure No 9 bacterial Count

 

Figure No 10 Control for bacterial Count

 

Figure No 11 Yeast and Mould Count

 

Figure No 12 Control for Yeast and Mould Count

 

Figure No 13 bacterial Count

 

Figure No 14 Control for bacterial Count

 

Figure No 15 Yeast and Mould Count

 

Figure No 16 Control for Yeast and Mould Count

 

CONCLUSION:

Nowadays herbal cosmetics are gaining popularity among customers there is a growing demand for herbal cosmetics in the world market and they are invaluable gifts of nature. The prepared poly herbal face cream was O/W type. Formulations C1 to C2 were prepared with the same ingredients but with different compositions of emulsifiers and thickeners. Both the cream formulations were observed to have similarity in consistency. There was no apparent change in the phase separation after centrifugation and physical appearance with formulation C2. But formulation C1 was found to be slightly unstable and contaminated so the formulation C1 was discontinued from the study. Therefore, C2 was chosen as the final formulation to prepare the cream for this thesis. Since both these plants Neem and Tulsi are well known for their anti-inflammatory, antioxidant and various other activities, we have chosen these two plants and tried to make a poly herbal face cream containing the extract of Azadirachtaindica and Ocimum sanctum. Our study indicated that the formulation C2 was found to be more stable, while the remaining formulation C1 was not stable and resulted in breakdown of the emulsion when stored for long time. The formulation C2 had almost constant pH, homogeneous, emollient, non-greasy and easily removed after the application. The stable formulations were safe in respect to skin irritation and allergic sensitization. Therefore to confirm the microbial stability of the formulation the cream was tested against various microorganisms the results revealed that no microbial growth was observed in the formulation. The obtained results had confirmed the microbial stability of the formulation.

 

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Received on 12.06.2018 Accepted on 28.06.2018

A&V Publications all right reserved

Research J. Topical and Cosmetic Sci. 9(1): Jan.-June 2018 page 25-32.

DOI:10.5958/2321-5844.2018.00006.7