Formulation and Evaluation of Antioxidant Face Cream Containing Raspberry Fruit and Grape Seeds Extract

 

Priya R. Kawarkhe*, Subhash V. Deshmane, Kailash R. Biyani

Department of Cosmetic technology, Anuradha College of Pharamcy, Chikhli. Dist-Buldana. 443201. M.S. India

*Corresponding Author E-mail: kawarkhepriya@gmail.com, subdeshmane@yahoo.co.in, principal.acpchikhli@gmail.com

 

ABSTRACT:

Antioxidant neutralizes free radicals, unstable oxygen molecules that break down skin cells and cause wrinkles, thus preventing impairment at the cellular level. In the present work, face cream formulations were prepared using antioxidant from natural source. Raspberry extract and Grape seed extract was used. Radical scavenging activity of raspberry, grape seeds was studied with standard ascorbic acid by DPPH method. Six cream base formulations were prepared and two formulations BF1 and BF4 were optimized. Six formulations of antioxidant cream were prepared using 2% extract. The various evaluation test like, pH, viscosity, grittiness, spreadability, skin irritation and antimicrobial test were carried out. The stability study over one week was also conducted. Raspberry and grapes seeds showed very good antioxidant activity with IC50 value 0.033µg/ml and 0.072µg/ml respectively. Face cream formulation shown active zone of inhibition within range of 9-12 mm, indicating that, all the formulation have good preservation activity. The study was concluded with that, natural source for excipient is the more suitable due to easy availability, non-toxicity and biocompatible properties of excipient. After studying the physical parameters, stability study evaluation, it is concluded that the product has the best capacity to reduce free radical and can be used as the provision of a barrier to protect skin.

 

KEYWORDS: Antioxidant, face cream, DPPH, raspberry fruit extract, grape seeds extract.

 


 

1. INTRODUCTION:

Skin facial creams restore a more youthful complexion, whitening the colour of skin, adds moisture and protective layer on the skin. The facial creams acts as removal of the flaws from skin, maintaining the smoothness and moistness of skin.1 The skin is the largest external organ of body and also known as the cutaneous membrane that protects against mechanical trauma, UV light and infections.2,3

 

It protects the underlying structures from injury and from invasion by microbes. Antioxidant neutralizes free radicals, unstable oxygen molecules that break down skin cells and cause wrinkles, thus preventing impairment at the cellular level. They inhibit inflammation leading to collagen depletion and offer protection against photo damage and skin cancer.4 It may also reduce the effect of trace, smoke, and pollution. Some skin expert believes that direct application of antioxidant cream to the skin might slow down the aging process.5 Now a day’s antioxidant obtained from natural herbal source is also have wide applications in preparation of cosmetic preparations because of their easy availability and non-toxicity.6 Raspberry, perennial plant species with woody stems in the genus Rubuscrataegifolius of the rose family.33 The fruit is an aggregate fruit 1 cm diameter, made up of numerous drupelets. The species grows on forest margins and mountain slopes, in areas with moist and well-drained soil. Red raspberry contains 31 μg/100 g of folate. Red raspberries have antioxidant effects that play a minor role in the killing of stomach and colon cancer cells. Grapeseeds extract, the extract obtained from the seeds of Vitisvinifera, belonging to the family vitaceae. Many of the other Vitisvinifera (grape) ingredients are reported to function as skin conditioning agents, and a few are reported to function as antioxidants.7

 

2. EXPERIMENTAL WORK:

Materials: Raspberry extract and Grape seed extract was procured from Kingvish Kompany, Thane, India. Other chemicals were used are of analytical grade.

 

2.1 Evaluation of Antioxidant activity of Raspberry and Grape seed Extract

2, 2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of the ethanol extract of raspberry and grape seed extracts in alone and combination was investigated. Stock solution of DPPH (100 mmol/L, 33mg in 1L) was prepared in methanol and labeled as control. The absorbance of control was measured using UV-Visible spectrophotometer at 517 nm. 5 ml of control as stock solution was added to 1ml of test sample dissolved in ethanol was added at different concentrations (50, 100, 200 μg/mL). Equal amount of ethanol was added to the control.8 After 30 min. absorbances of all three test samples were measured at 517 nm spectophotometrically 9. The scavenging activity was calculated using the formula:

 

% Scavenging activity

= [(A517control-A517sample)/A517control] x 100.

Control = stock solution of DPPH, Ascorbic acid was used as a standard.

 

2.2 Formulation of cream base

Ingredients were used in the preparation of cream base are mentioned in table 1. The base was prepared by emulsification method.

 

2.3 Evaluation of cream bases

The six formulations of cream base were subjected to various evaluation tests. The pH of base was determined by pH meter. The viscosity of cream bases formulation was determined by using Brookfield viscometer at 100 r/min. Spreadability of formulations were determined by fabricated apparatus in laboratory and used for study. The apparatus consist of wooden block, with a fixed glass slide with one end tied to weight pan rolled on the pulley, which was in horizontal level with fixed slide. 1.5gm was placed between two glass slide and 100gm weight was placed on slide for 5 min to compress. The sample to uniform thickness weigh was added to the pan. The time (seconds) required to separate the two slides was taken as a measure of spreadability. It was calculated using the formula:

              

Where,

m = weight tide to upper slide, l = length moved on the glass slide, and t = time taken (in sec) to separate the slide completely each other.

 

Skin irritancy test was conducted using six human volunteers. An area of 1 cm2 on the dorsal left hand surface was marked. The cream was applied to the specified area and the time was noted. Irritancy, erythema, edema were checked for regular intervals up to 24 h and reported. The stability study of cream base formulations at the room temperature was carried out for three weeks and all above parameters were checked. The ease of removal of the cream applied was examined by washing the applied part with tap water.

 

2.4 Preparation of antioxidant face cream

Antioxidant face cream containing raspberry (RB) and grape seed (GS) extract was prepared with optimized base cream formulation. The details of components of face cream are mentioned in table 2. The cream was prepared by emulsification method and water was used as vehicle up to the volume. The weight of cream was 100 g.

 

 


 

Table 1: Formulation of cream base

Formulation code

SA*

(g)

AO*

(ml)

NaOH

(g)

TEA*

(ml)

EDTA*

(g)

Glycerin

(ml)

MP*

(g)

Perfume

(ml)

BF1

13

1

1

1

0.2

6

0.1

0.5

BF2

14

1

1

1

0.2

6

0.1

0.5

BF3

15

0.5

1

0.5

0.2

8

0.1

0.5

BF4

16

0.5

1

0.5

0.2

8

0.1

0.5

BF5

17

0.5

1

0.5

0.2

8

0.1

0.5

BF6

18

0.5

1

0.5

0.2

8

0.1

0.5

* SA- Stearic acid, AO- Almond oil, TEA- Triethanolamine, EDTA-ethylenediamine tetra acetic acid, MP-Methyl paraben

Table 2: Composition of face cream containing active ingredient

Formulation code

Fruit extract

(2 %)

SA*

(g)

AO*

(ml)

NaOH

(g)

TEA*

(ml)

EDTA*

(g)

Glycerin

(ml)

MP*

(g)

Perfume

(ml)

AF1

RB

14

0.5

1

0.5

0.2

8

0.1

0.5

AF2

GS

14

0.5

1

0.5

0.2

8

0.1

0.5

AF3

RB + GS

14

0.5

1

0.5

0.2

8

0.1

0.5

AF4

RB

15

0.5

1

0.5

0.2

8

0.1

0.5

AF5

GS

15

0.5

1

0.5

0.2

8

0.1

0.5

AF6

RB + GS

15

0.5

1

0.5

0.2

8

0.1

0.5

* SA- Stearic acid, AO- Almond oil, TEA- Triethanolamine, EDTA-ethylenediamine tetra acetic acid, MP-Methyl paraben

 

 

 


2.5 Evaluation test for antioxidant face cream

The all formulations of antioxidant cream were subjected to various primary evaluation tests like pH, viscosity, spreadability and grittiness, skin irritancy, color and washability. The anti-microbial assay test for the antioxidant cream formulations  was carried out using disk diffusion method  against two human pathogenic bacteria, including Gram positive (Staphylococcus aureus), Gram-negative bacteria (E. Coli). The cultures were grown in nutrient broth and incubated at 37 °C for 24 h. A sterile 6mm diameter filter discs (whatman paper no. 42) impregnated in 6 batches of cream formulation placed on the surface of petri plates seeded with 0.1 ml of microbial suspension. Soon afterwards the plates were kept at 10oC for 30 min. After it normalized to room temperature the plates were incubated at 37 şC for 24 h. After incubation period the zone of inhibition was measured.13 The stability study at room temperature was determined for the period of 3 week. The tests were studied was color, sudden viscosity change and feel properties 10-12.

 

3. RESULT AND DISCUSSION:

FTIR spectrums of Raspberry, Grape seed extract and combined Raspberry and Grape seed extract are mentioned in figure no.1, it was found that the all characteristic functional group were found between specified band widths and hence identified pure. There is a slight shifting of peaks, but that is also in specified ranges, hence found both extract was compatible with each other.


 

 

 

Figure 1: FTIR study of Raspberry fruit, grape seeds extract and mixture of raspberry and grape seeds

 

 

 


3.1   Antioxidant activity

The DPPH as a control was measured with absorbance 0.0775. The highest percent inhibition of Raspberry fruit extract was found 94.83 % with IC50 value 0.033 µg/ml. Grape seed extract showed 90.96 % inhibition activity with IC50 value 0.072 µg/ml. While, ascorbic acid gives percent inhibition activity was 89.67% with IC50 value 0.150 µg/ml.  The combination of Grape seed and Raspberry extract has 87.09 % inhibition with IC50 value 0.161 µg/ml. The different concentrations showed different percent inhibition which were mentioned in table no. 3.

 


 

 

Table 3: Absorbance and % Inhibition activity of raspberry extract.

Substance

Concentration (μg/ml)

Sample absorbance

% Inhibition

IC50value

(μg/ml)

Raspberry extract

50

0.018

76.77 %

0.148

100

0.014

81.93%

0.115

200

0.004

94.83%

0.033

Grape seed extract

50

0.019

75.48%

0.195

100

0.012

84.51%

0.123

200

0.007

90.96%

0.072

Ascorbic acid

50

0.014

81.93%

0.264

100

0.013

83.22%

0.245

200

0.008

89.67%

0.150

Raspberry and grape seed extract

50

0.017

78.06%

0.274

100

0.016

79.35%

0.258

200

0.010

87.09%

0.161

 

 

 

                                                           

 

 

Figure 2: Graph showing antioxidant activity

 

 

 

Table 4:  Evaluation of different parameters of base formulations

Formulation code

pH

Viscosity

(cps)

Spreadability

(cm/sec)

Skin irritation

BF1

5.8

32533

29±1

No Irritation

BF2

6.2

34753

27±1

No Irritation

BF3

6.1

40200

26.33±1.52

No Irritation

BF4

5.9

41350

25±1

No Irritation

BF5

6.8

52400

23.66±1.5

No Irritation

BF6

6.9

55240

22.66±1.52

No Irritation

 

 


 

Figure 2, indicated that single Raspberry fruit extract of 200 µg/ml concentration showed highest percent free radical scavenging activity (94.83%) as compare to 200 µg/ml ascorbic acid (89.67%). From the observation (fig. 4,5,6 and 7) it was notice that single Raspberry fruit extract having good antioxidant activity than Grape seed extract or combined form of Raspberry and Grape seed extract. Also it was observed that Grape seed extract and combine extract show antioxidant activity near to ascorbic acid. Hence, both extracts were considered for formulation of herbal antioxidant face cream.

 

3.2 Evaluation of base formulations

Six formulation of primary base were prepared by melting method satisfactorily. All the formulation was subjected to various evaluation parameters. The results of various tests are mentioned in table 4 and found satisfactory. All formulations were white in color, feel was very smooth, viscosity was acceptable and no irritation was cause.

 

Form the data mentioned in table 4, it was found that except formulation no. BF5 and BF6 all formulations were shown desire pH (Desire pH should be 4.5 to 6.2). The viscosity of all formulations was also passable with good spreadability (22.6-29 cm/s). All formulations were non-irritant. Remaining parameters were good and/ or desirable, hence at this stage formulations BF1 to BF4 were found good stable.  After 3 week stability study at room temperature all the cream base formulations were stable upto 1 week. Afterwards slight change in viscosity and feel was found with formulation BF5 and BF6 respectively. It might be due to increasing concentration of stearic acid. In 3rd week only formulation BF2 and BF4 was found stable with white colour, smooth feel and a very specific pH and viscosity (Table 5). Hence, for the preparation of Antioxidant Face Cream we considered formulations BF2 and BF4 as optimized batches. This may be due to appropriate concentration of stearic acid.

 

 


 

Table 5: Stability study of base formulation

Formulation code

Color

Sudden viscosity change

Feel

pH

Viscosity (cps)

After 1 Week

BF1

White

No change

Smooth

5.7

32630

BF2

White

No change

Smooth

6.4

34770

BF3

White

No change

Smooth

6.1

40213

BF4

White

No change

Smooth

6.2

41355

BF5

White

No change

Smooth

6.5

52450

BF6

White

No change

Smooth

6.9

55260

After 2 Weeks

BF1

White

No change

Smooth

5.7

32726

BF2

White

No change

Smooth

6.1

34793

BF3

White

No change

Smooth

6.0

40225

BF4

White

No change

Smooth

5.8

41350

BF5

White

No change

Smooth

6.6

52560

BF6

White

Change

Tacky

6.5

55467

After 3 weeks

BF1

White

Change

Smooth

6.4

32742

BF2

White

Change

Smooth

5.8

36730

BF3

White

No change

Smooth

6.1

45220

BF4

White

No change

Smooth

5.9

46358

BF5

Yellowish white

Change

Tacky

6.4

53577

BF6

Yellowish white

Change

Tacky

6.6

56489

 

 

Table 6: Evaluation of different parameters of cream formulations

Formulation code

pH

Viscosity

(cps)

Spreadability

(cm/sec)

Skin irritation

Zone of inhibition

AF1

6.2

38590

32.6±0.55

No Irritation

9 mm

AF2

6.5

38166

31.21±0.93

No Irritation

12 mm

AF3

6.9

37459

27.96±0.40

No Irritation

9 mm

AF4

6.6

45470

24.3±0.95

No Irritation

7 mm

AF5

6.8

46598

26.3±0.95

No Irritation

9 mm

AF6

6.9

46577

21.42±0.87

No Irritation

11 mm


 

 

3.3 Evaluation of antioxidant cream formulations

Results shown in table 6, indicated that all the cream formulations were found within the acceptable limit. The zone of inhibition was found within range of 9-12 mm, indicating that, all the formulation have good preservation activity. Stability tests at room temperature for cream formulation for 3 weeks were conducted satisfactory and the results were shown in table no.7. Among all the formulas containing active AF1 showed good consistency and pH around 6 up to 3 week study and it is also suitable for topical application as the pH of the skin is in between 4.5 to 6.2.

 


 

 

 

Table 7: Stability study of cream formulations

Formulation code

Color

Sudden viscosity change

Feel

pH

Viscosity

(cps)

After 1 week

AF1

White

No change

Smooth

6.2

38578

AF2

Faint brown

No change

Smooth

5.8

38234

AF3

Off white

No change

Smooth

6.6

37467

AF4

White

No change

Smooth

6.5

45477

AF5

Faint brown

No Change

Smooth

6.8

45892

AF6

Off white

No Change

Smooth

6.9

45077

 After 2 weeks

AF1

White

No change

Smooth

5.9

37560

AF2

White

Change

Tacki

6.4

38679

AF3

Off white

No change

Smooth

6.8

38489

AF4

White

No change

Smooth

6.5

45588

AF5

Faint brown

Change

Tacki

6.7

48789

AF6

Off white

No change

Smooth

6.5

45578

After 3 weeks

AF1

White

No change

Smooth

6.1

45768

AF2

White

Change

Tacki

6.6

38457

AF3

Off white

No change

Smooth

6.5

39467

AF4

White

No change

Smooth

6.3

47855

AF5

White

Change

Tacky

6.8

48765

AF6

Off white

Change

Tacky

6.6

49521

 

 

 


4. CONCLUSION:

Raspberry fruit extract and grape seed extract showed very good free radical scavenging (antioxidant) activity. Stearic acid, triethanolamine and other ingredients were used in preparation of face cream also prove stability of the preparations. Natural source for excipient is the more suitable due to easy availability, non-toxicity and biocompatible properties of excipient. After studying the physical parameters, stability study evaluation, it is concluded that the product has the best capacity to reduce free radical and can be used as the provision of a barrier to protect skin.

 

5. REFERENCES:

1.     Anne Waugh, Allison Grant, Ross and Wilson Anatomy and Physiology, Churchill Livingstone Elsevier, 11(1), 2009:353-358.

2.     Gerard J. Tortora Bryan, Derrickson, Principles of Anatomy And Physiology, John Wiley And Sons Pvt. Ltd.13(1),2011:154-165.

3.     Rahul Phate, Anatomy, physiology and health education, career publication, 2008;3(1):244-247.

4.     Christian Oresajo, Sreekumar Pillai, Margarita Yatskayer, Germain Puccetti,  David H. McDaniel, Antioxidants and Skin Aging: A Review Cosmetic Dermatology 2009;22(11):563-570.

5.     Abhay Prakash Mishra, Formulation and evaluation of herbal antioxidant face cream of Nardostachys jatamansi collected from Indian Himalayan region, Asian Pacific Journal of Tropical Biomedicine, 2014; 4(2): S679-S682.

6.     Dragana Stojiljković, Oxidative Stress, Skin Aging and Antioxidant Therapy, Scientific Journal of the Faculty of Medicine in Niš 2014; 31(4):207-217.

7.     Chair, Wilma F. Bergfeld,  Donald V. Belsito, Ronald A. Hill, Curtis D. Klaassen,  Daniel C. Liebler, James G. Marks, Jr., Ronald C. Shank,  Thomas J. Slaga, ; and Paul W. Snyder, Safety Assessment of Vitis Vinifera (Grape)-Derived Ingredients as Used in Cosmetics, Cosmetic Ingredient Review; 2012:1-35.

8.     Warrier, P.K, Nambier, VPK, Raman Kutty C, Indian medicinal plants- A compendium of 500 species, Orient Longman Ltd, Madras, 1994, Vol-I, 95-97.

9.     Harborne, J.B, Phytochemical methods- A guide to modern techniques of plant analysis, 3rd Edn, Springer (India) Pvt. Ltd, New delhi,1998, 5-32.

10.   Indian Standards, Specification for Skin Cream. 2001, 3 -10

11.   Cleidson Valgas: Simone Machado De Souza; Elza FaSmania; Artur Samania Jr, Screening method to determine antibacterial activity of natural product, Brazilian Journal of Microbiology (2007).

12.   Sujith S Nair, Molly Mathew and Sreena K., Evaluation of skin irritation of herbal antioxidant cream, Formulation of Some Antioxidant Herbal Creams. Asian Journal of Biochemical and Pharmaceutical Research 2012; 3(2):184-188.

 

 

 

 

 

Received on 10.11.2016                    Accepted on 20.12.2016  

©A&V Publications all right reserved

Research J. Topical and Cosmetic Sci. 2016; 7(2): 73-78.

DOI: 10.5958/2321-5844.2016.00012.1