IN VITRO EVALUTION OF SHAMPOO

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K. JAYA RAJ KUMAR Dept.pharmaceutics Rao’s college of pharmacy. Nellore. [email protected] IN VITRO EVALUATION OF ANTI DANDRUFF SHAMPOO

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METHODS FOAM AND FOAM STABILITY The Ross-Miles foam column method used measure foam height and stability. In this test 200 ml of 0.1% liquid shampoo solution falls through an orifice into a glass column containing 500 cc of the same solution. After a specified period of time at once and after five minutes, the height of foam is measured.

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Cleansing power is evaluated by the method of barnet and powers, 5gm sample of soiled human hair is placed at 35oc in 200 cc of water containing of 1 gm of shampoo. The flask is shaken 50 times a minute for 4 minutes. Then washed once again with sufficient amount of water, then after filter the hair dried and weighed. The amount of soil is removed under these condition is calculated. CLEANING ACTION:

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Uniform size of human hairs was collected. Then hairs washed with sufficient amount of water and dried. The Accurate amount of hair (5g) soiled with standard soil then weighed. These hairs taken for cleaning action. COLLECTION OF HAIR: PREPARATION OF STANDARD SOIL Combination of carbon black and mineral oil.

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Canvas disk sinking test: A mount veron cotton duck # 6 canvas disk 1 inch in diameter, is floated on the surface of a solution, and the time required for it to sink is measured accurately. WETTING ACTION:

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The wells are dig on agar plates with sterilised well digger aseptically. Take 100µml of each sample, add to well aseptically. Incubate the plates at 37oC for 24 hrs to 48 hrs. Observe the effectiveness of sample on culture growing on the agar plate and we can see the effectiveness of sample in the form of zone of inhibition around each well containing different sample. MAKING THE WELLS ON AGAR PLATES

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Conditioning action is a difficult property to assess. This is because it is basically dependent on subjective appraisal. No method has been published for measuring conditioning action. The degree of conditioning given to hair is ultimately judged by shampoo user who is making the evaluation on the basis of past experience, present expectations, and continuing change is the individual scalp and hair situation. Conditioned hair should be soft, lustrous, easily combined and coiffures. CONDITIONING ACTION:

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EVALUATION OF CONDITIONING ACTION: The prepared shampoo formulations were given to volunteers and they were asked to give their opinion, on the conditioning ability of preparation. After two weeks volunteers were of the opinion that the shampoo gives the hair a lustrous look, softness and hair is easily styled.

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VISCOSITY: Viscosity of the liquid shampoo was determined using a Brooke field 100 mL of the shampoo was taken in a beaker and the spindle was dipped in it for about 5 min and then the reading was taken. MICROBIAL TEST: PREPARATION OF PRE-INOCULUM Take the loopful culture of staphylococcus aureus (ATCC6532) aseptically and transfer to sterilized and cooled 100 ml SCDM (broth).Mix well. Incubate the broth at 37oC for 24 hrs.

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PREPARATION OF MEDIA Soya bean casein digests medium, soya bean casein digest agar and nutrient agar. PREPARATION OF POUR PLATES Sterilized SCD agar (100 ml) is cooled to 40oC and mixed with 5 ml of 24n hrs old pre inoculated culture. This is immediately poured in plates (340 ml each) and allows to set.

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MAKING THE WELLS ON AGAR PLATES The wells are dig on agar plates with sterilised well digger aseptically. Take 100µml of each sample, add to well aseptically. Incubate the plates at 37oC for 24 hrs to 48 hrs. Observe the effectiveness of sample on culture growing on the agar plate and we can see the effectiveness of sample in the form of zone of inhibition around each well containing different sample.

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The test calls for dropping 0.1 ml of liquid shampoo the test substance in the conjunctiva sac of one eye of the rabbit , the other eye serving as control. In the case of the first three animals, the treated eye remains unwashed. Since washing the eye may or may not alleviate symptoms of injury. The six remaining animals are divided into two equal groups in the first of these groups eyes instilled with the substances are washed with 20 ml of lukewarm water two seconds after treatment and in the second group after instillation. The washing is regarded as significant since it is important to know the effect of such a procedure that is, whether it is detrimental or beneficial and if beneficial, to ascertain the extent of the benefit of the made at 24, 48 and 72 hr and again four and seven days after treatment. If the lesions have not cleared up in seven days the test material is considered as severe irritant. EYE IRRITANCY TEST:

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REFERENCE: Edward Sagarin, Cosmetics Science and Technology, Page No. 409, 641 to 646, 403 to 407,414, 419. Balsam, Sagarim, “Cosmetics Science Technology” Volume 2, 2nd edition, page No. 105 to 112. Poucher, “Perfumes cosmetics and soap and volume 3, page No. 108 to 109, 121 to 123, 110. Ross miles (Miles, G.D. Ross, J and Shed Lovsky, L.J. Am Oil Chemists Soc. 27, 27-273). Barnett G. And Powers, D.H. “The effect of soft water, hard water and sea water on the performance of shampoos and surface active agents Proc. Sci., Tern, 15:16(1951).

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Thank You

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