Research article SKIN REJUVENATING SERUM WITH KAKADU PLUM EXTRACT AS AN ACTIVE

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1 Research article SKIN REJUVENATING SERUM WITH KAKADU PLUM EXTRACT AS AN ACTIVE Nitesh Rajput 1 Vishal Chinchamalatpure 2* 1 Senoir Manager,R & D, Vitro Med Health Care, Jaipur. 2 Executive, R & D Cosmetic, Sri Sri Ayurveda Trust, Bangalore *Corresponding author id: vishalchinchamalatpure@gmail.com Abstract: Skin aging is the severe problem faced by the people these. According to the literature survey it is studied that Kakadu Plum Contains Vitamin C Vitamin C is a highly effective antioxidant and acts to lessen oxidative stress. At first the active was selected for skin Rejuvenating product. Then the excipients were selected for the Base formulation. New excipients like Hydroxy ethyl cellulose, glyceryl stearate, glyceryl cocoate were used to give the additional stability to the formulation. Various Base formulations were prepared by variation in excepients. Optimization of these various Base formulations was done by considering the physical parameters like appearance, colour, consistency, thermal stability, feel and Spreadability. Then the in-vitro evaluation of Serum Base was performed. Satisfactory Base formulation was selected among these formulations by comparing them on the basis of optimization and in-vitro evaluation of these Serum Bases. The various concentrations of Kakadu Plum Extract 1%, 2%, 3% & 5% were incorporated in the selected Serum. The Serum was prepared by conventional procedure. Optimization of these formulations containing 1%, 2%, 3% & 5% of Kakadu Plum Extract was done by considering the physical parameters like appearance, color, consistency, thermal stability, feel and Spread ability. It was kept stability testing for 21. Here these 2% & 3% formulations were observed after each interval of 7 for 21. In the stability studies ph is observed. And it was found that both the formulations were stable and had no changes in ph. Keywords: Kakadu Plum extract, vitamin C, base serum, thermal stability INTRODUCTION: Skin products are becoming increasingly popular and in high demand all over the world. Some women often express a desire to lighten skin tone by achieving improved visible tone, reduction in yellowness (or sallow tone), and reduction in the appearance of hyper pigmented spots ( age or sun spots). existing transdermal dosage forms. The dark or pigmented skin is treated with skin rejuvenating cosmetic product containing new Kakad Plum Extract an active. Drug profile: Kakadu Plum (S Mohanty, Ian E Cock, 2012). Synonyms : Terminalia ferdinndiana. Final report of the safety assessment of L-Ascorbic Acid, Calcium Ascorbate, Magnesium Ascorbate, Magnesium Ascorbyl Phosphate, Sodium Ascorbate, and Sodium Ascorbyl Phosphate as used in cosmetics. (Elmore, 2005). The objective of work is to develop and investigate a novel skin product for the pigmented skin as alternative to the Description: Terminalia ferdinandiana is a slender, small to medium-sized tree growing up to 32 m in height. Large shrub or small tree, m tall; stems spreading, bark grayish. Serumy-grey, flaky bark and deciduous pale green leaves. The flowers are small, Serumy-white, perfumed, and borne along spikes in the leaf axils towards the ends of the branches. Flowering is from September to December. (Southern

2 hemisphere spring / summer). The fruit is yellow-green, about 2 centimetres long and 1 centimetre in diameter, almond-sized with a short beak at the tip, and contain one large. They ripen from March onwards. Part used: fruits MATERIAL AND METHOD 1. Mineral Oil 2. Cetyl alcohol 3. Glyceryl Monostearate 4. Isopropyl Myristate 5. Propylene glycol 6. Kakadu Plum Extract. Table. 1: Optimization of Serum Base: 7. Citric Acid- 8. Perfume 9. DMDM Hydantoin 10. Cyclopentasiloxane 11. Hydroxy Ethyl Cellulose PROCEDURE: (Murad Alam et al., 2009) Clean the apparatus, weight the ingredients as per formulation F1, F2, F3, F4.weight separately oil phase and water phase. Heat both the phase up to 80 0 C. Add oil phase to water phase and thus emulsification process was carried out with the help of mechanical stirrer. When temperature lowers down to 40 0 C then perfume was added. All the formulations were kept for 48 hours stability test. Sr.. Parameter F1 F2 F3 F3 1 Appearance Colour Odor Consistency Feel Spedability Here, + = Good, ++ = Better, +++ = Best From the above observation formula F2 was stable & it shows consistency, spreadability and feel, therefore it was selected and extract was added with different concentration and forward for in vitro study as per IS and in vivo study with human volunteers. Analysis of Skin Rejuvenating Serum In vitro studies: The prepared Skin Rejuvenating Serum with Kakdu Plum Extract as an Active: Evaluation of serum Determination of Physical Parameters: (S Mohanty, Ian E Cock, 2012) In physical parameters, appearance, consistency, color, odor, and spredability was taken into consideration. Procedure: The Physical Parameters are determined by visual observation by taking small amounts of sample. The Serum and lotion samples were kept at various temperatures such as room temp, at 45 C and at the elevated temp (freeze temp.).the formulations were checked after every 10 and parameters Such as color, odor, consistency, spreadabilty and appearance. Determination of ph: (Ghulam Murtaza et al., 2010). Procedure: For oil-in-water emulsion Serum Accurately 5±0.01 g of the Serum was weighted in a 100ml beaker. 45ml of water was added and the Serum was dispersed in it. The ph of the suspension at 27 0 C was determined using the ph meter. Determination of total fatty substance content: (Indian Standard skin creams specification, 2004) Principle of the method: The emulsion is broken up with dilute mineral acid and the fatty matter is extracted with petroleum ether. It is weighed after removal of the solvent. Procedure: Accurately about 2g of sample was weighted into a conical flask, about 25ml of dil HCl was added, reflux condenser was fitted into the flask and the content of the flask was boiled until the oil and water phases have separated. The content of the flask was poured into 300 ml separating funnel

3 and it was allowed to cool to 20 0 C. The conical flask was rinsed with 50ml of ethyl ether in portions of 10ml. The ether rinsing was poured into separating funnel. The separating funnel was shaking well and leave until layers separate. Separate out with 50ml portions of ether twice. All the ether extracts was combined and washed them with water until free of acid. The ether extracts was filtered through a filter paper containing sodium sulphate into a conical flask which had been previously dried at temperature of 60±2 0 C and then weighed. The sodium sulphate was washed on the filter paper with ether and the material remaining in the flask was dried at a temperature of 60±2 0 C to constant mass. Calculation Toal fatty substance % by mass =100x M1/M2. Where, M1 is Mass in g of residue and M2 is Mass in g of material taken for the test. Determination of Thermal Stability: (W.A. Robinson and M. Katouli, 2006) Procedure: A 20 mm broad and 5 mm thick strip was spreaded from the material to be tested on the internal wall of a beaker of 100ml capacity in its total height. The beaker was kept for 8 hrs. in the humidity chamber at 60 to 70% relative humidity and temperature 37±1 0 C. Microbial examination of Serum: T. (Wohlsen et al., 2006) Outline: The test consist of pleating a known dilution of the sample on soya bean casein digest agar medium suitable for the total count of aerobic bacteria and fungi after incubating them for a specified period to permit the development visual colonies for counting Table. 2: Stability studies of Serum Pretreatment of sample: To 10 ml of sodium chloride solution ph 7 or any other suitable medium add 1gm or 1ml of sample Total bacterial count: Pipette out in duplicate 1ml of pretreated sample aseptically into 5 sterile petridishes. Pour 15 to 20 ml of molten soya bean casein digest agar maintained at about 45 0 C. Mix the content of the plate by swirling. Allowing the incubate the plates at 37 0 C C in inverted position for three Count the number of colonies in each plate. Determine the average number of colonies on plates and multiply by dilution factor. This will be the number of microorganisms per gm of the sample. If no colony was recovered from any of the plate it can be stated as less than 50 microorganisms per gm Total fungal count:pipette out in duplicate 1ml of pretreated sample aseptically into 5 sterile petridishes. Pour 15 to 20 ml of molten sabouraud s chloranphenicol agar (SCA) maintained at about 45 0 C mix the content of the plate by swirling. Allowing the plates to solidify, invert and incubated at C for three. Count the number of colonies in each plate. Stability studies of Serum: (Ghulam Murtaza et al., 2010) Stability studies for Serums were carried out according to ICH guidelines. The Serum samples were kept on the 5 0 C, room temperature, and 40 0 C. The changes in the physical appearance, color, odour etc and chemical changes such as change in ph, viscosity, ph separation were checked and thus. The formulation of Serum was optimized. Sr.. Parameters Skin Rejuvenating Serum containing Kakadu Plum Extract as an Active A) Physical parameter F1 F2 F3 1 Appearance Serum like Serum like Serum like 2 Color Light Light Light 3 Consistency Semi liquid Semi liquid Semi liquid 4 Spreadability Good good Very Good 5 Oil / tacky feel

4 Accelerated stability studies: To ensure that a cosmetic remain stable till the consumers has used the entire cosmetic or has stopped using it, a number of special accelerated test procedures have been developed. The evaluation employs a combination of tests. This method of evaluation not only indicates stability of Base formulation but also indicates the stability of functional ingredient. Cyclical temperature tests (S Mohanty, Ian E Cock, 2012): These tests are not carried out at fixed temperature and humidity. In these tests, temperature was changed cyclically every day eg. Low-high-low-high-low-high, to simulate changes in temperature daily. Table. 3: Cyclical temperature tests for serum. Parameters Skin Rejuvenating Serum containing Kakadu Plum Extract as an Active Cyclic Parameter F1 F2 F3 1 Low-high-low-high number of cycles 5 from 1-20 Stable Stable Stable RESULT AND DISCUSSION: In Vitro-Study Table. 4: Determination of physical parameter of a Skin Rejuvenating Serum containing Kakadu Plum Active extract. (Stability study after 10 ) Characteristic 1 Appearance NC NC NC NC 2 Color NC NC NC NC 3 Odor NC NC NC NC 4 Consistency NC NC NC NC 5 Spredability NC NC NC NC Table. 5: Stability study after 20 5 Spredability NC NC NC NC Table. 6: Stability study after 30 Characteristic Characteristic 1 Appearance NC NC NC NC 2 Color NC NC NC NC 3 Odor NC NC NC NC 4 Consistency NC NC NC NC 1 Appearance NC NC NC NC 2 Color NC NC NC NC 3 Odor NC NC NC NC

5 4 Consistency NC NC NC NC 5 Spredability NC NC NC NC Table. 7: Determination of ph of serum incorporated with Kakadu Plum Extract. Standard value 5 to 9 Sr. Time Interval 1 0 day th day th day th day th day Graph F1 F2 F3 F Time Interval Graphical Representation ph Skin Rejuvenating Serum containing Kakadu Plum Extract Active Table. 8: Determination of Total Fatty Matter of : Standerd Value : 3 Min Sr. Characteristics 1 TFM Percent by Mass 4.4% 4.55% 4.5% 4.44%

6 Graph. 2 % By Mass Total Fatty Matter F1 F2 F3 F4 Graphical Representation of Determination of Total Fatty Matter of Skin Rejuvenating Serum containing Kakadu Plum Extract as an Active Table. 9: Determination of Thermal Stability of : Sr.. Parameters 1 Thermal stability Passed Passed Passed Passed Microbial examination of a Serum Table. 10: Microbial examination of a Serum containing Kakadu Plum Extract. Sr. Name of test Result specification unit 1 Total bacterial count 10 CFU/gm NMT100 CFU/gm CFU/gm 2 Fungal count NIL NMT10 CFU/gm CFU/gm The Total bacterial count Of a Serum containing Kakadu Plum Extract was found to be 10 CFU/gm that is <100 CFU/gm therefore the Serum passes the test. The fungal count of a Serum containing Kakadu Plum Extract was found to be NIL. Therefore the Serum passes the test CONCLUSION Wrinkles, fine lines and uneven skin texture are persistent and universal concerns of aging patients, particularly for women. Skin Rejuvenating products are becoming increasingly

7 popular and in high demand all over the world. From the above study the skin Rejuvenating product is developed using Kakadu Plum Extract a vitamin C derivative. This skin rejuvenating product gives satisfactory skin Rejuvenating property. This Kakadu Plum Extract is used as derivative of vitamin C which is responsible for healthy skin. This product not only lightens the skin but also it keeps the skin healthy and fresh. This quality of Kakadu Plum Extract is lack in other skin lighteners. And therefore the skin rejuvenating product using Kakadu Plum Extract was developed.therefore it is suggested that the prepared formulation (o/w type Serum) containing 3% Kakadu Plum Extract can be used to increase the Rejuvenation of the skin. REFERENCE: Elmore AR (2005).Final report of the safety assessment of L- Ascorbic Acid, Calcium Ascorbate, Magnesium Ascorbate, Magnesium Ascorbyl Phosphate, Sodium Ascorbate, and Sodium Ascorbyl Phosphate as used in cosmetics.int J Toxicol. 24(2): Ghulam Murtaza, Muhammad K. Waqas, Naveed Akhtar, Mahmood Ahmad (2010).Formulation and characterization of a cream containing extract of fenugreek seed,acta Polonisae Pharmaceutica-drug Research,vol.67 2 pp ,. Indian standard skin creams specificationis (2004). 6608: Skin Creams [PCD 19: Cosmetics]. Bureau of indian standards manak bhavan, 9 bahadur shah zafar margnew delhi nd edition. Pp 3-4 Murad Alam, Ashish Bhatia, Rupal Kundu, Simon S.Yoo (2009). Cosmetic Dermatology for Skin of Color.pp-24. ISBN: S Mohanty, Ian E Cock (2012).The chemotherapeutic potential of Terminalia ferdinandiana: Phytochemistry. 6(11): T. Wohlsen, J. Bates, G. Vesey, W.A. Robinson and M. Katouli (2006). Evaluation of the methods for enumerating coliform bacteria from water samples using precise reference standards. 42: Source of Support: Nil Conflict of Interest: ne Declared