PATCH TESTING

Points to be studied
Principle behind patch test Types of patch testing General procedure involved Open & prophetic patch test Constraints in patch test Report of patch test

What is patch testing?

Patch testing is a way of identifying whether a substance that comes in contact with the skin is causing inflammation of the skin (contact dermatitis). There are two types of contact dermatitis:
irritant contact dermatitis and allergic contact dermatitis.

Irritant contact dermatitis

An irritant substance is one that would cause inflammation in almost every individual if it was applied in sufficiently high concentration for long enough. An irritant reaction is caused by the direct contact of an irritant substance with the skin and does not involve the immune system.

Allergic contact dermatitis

An allergic reaction is specific to the individual and to a substance (or a group of related substances) called an allergen. Allergy is a hypersensitivity to a particular substance, and always involves the immune system. All areas of skin that are in contact with the allergen develop the rash. The rash will disappear if you avoid contact with the substance.

Purpose of Patch Testing

Patch testing can help to differentiate between the two. The test involves the application of various test substances to the skin under adhesive tape that are then left in place for 48 hours. The skin is then examined further 48 hours later for any response. This can help the doctor/ cosmetic chemist, to decide which allergens you are allergic to and identify those that could be aggravating your dermatitis. The doctor will then be able to advise how you can avoid the allergens.

Types of patch test

Diagnostic patch test Prophetic patch test

Diagnostic patch test

The cosmetic which used, has caused dermatitis and if cosmetic is known, the ingredient which has caused it.

Prophetic patch test

For new cosmetic products. Whether new product should place in market or not.

General procedure
Place 0.1 to 0.3 gm of cosmetic to be tested on a piece of cotton or flannel (2-3 cm2) Apply to the skin of arms, thighs or back. This patch is covered with the patch of cellophane (5 cm2) and sealed with adhesive plaster (40 cm2) Apply several patches at one time. Some patches: similar cosmetics with other brand but known not to cause any harm to the skin. Other similar cosmetics: Controls Allowed to remain that patches for 24 72 hrs.

If there is no reaction, the same patch should reapplied to the same place or fresh patch of the same material may be made & applied. This may be continued till:
either a reaction is produced under one or more patches, or investigator is confirmed that no reaction will occur.

Reading & grading of patches: experienced dermatologist. Sites of the patches, examined after 30 minutes of removal of patch. Observation should not be done before 15 minutes of patch removal.
Skin under adhesive tape: inflamed, Skin under cellophane tape: clear Skin under test patch: no reaction or reactions like erythema, erythema with papules, papulory vascular reaction or ulceration or neurosis.

Erythema: redness of the skin caused by capillary congestion. Erythema with papules: elevated, solid lesion, less than 1 cm. Papulory vascular reaction: injury to blood vessels. Ulceration: inflammatory or more seriously, infective condition of the cornea involving disruption of its epithelial layer. Necrosis: premature death of cells and living tissues.

Sr. No
1 2 3 4 5

Description of observation Symbol

No reaction Erythema only Erythema with papules Papilovascular Ulceration or Necrosis + ++ +++ ++++

If no reactions are observed, the subjects should be observed for next 3 to 5 days bcoz some ingredients show late reaction. The skin is exposed to sunlight and find out, it causes any photosensitization.
For particular ingredients: Eighteen patches can placed on one subject:
Three on each arm & thigh and Six on the back.

OPEN PATCH TEST

Used for cosmetics containing higher % of potential irritants like hair dyes, shampoos, hair tonics. Sealing of patches does not simulate actual use so patches are kept open and performed on sensitive part of skin e.g. bend of elbow, popliteal space of the skin behind the ears. Cosmetics applied on 1 cm2 area. Control patches of similar cosmetic but well known brand applied. Site of patch inspected after 24 hrs. If no reaction: repeated once more on same site upto three times. If no reaction after third time: not hypersensitive If reaction occurs: hypersensitive.

PROPHETIC PATCH TEST

Before performing this test, investigator should confirm that the material is not the primary irritant. Careful about the sign of the subjects for the waiver of damage and money matter. 10 to 200 subjects can be used. One to five days for observation, three more days for late reaction. After 7 to 10 days, patches are reapplied for the same test if not shown any positive response in first. If there are no reactions or one or two reactions out of 200, new product can be placed for trial sale in a community of about 10,000 persons.

Prophetic test may be open or covered. Safe cosmetics: if more than 2 in 10,000 shows hypersensitivity reaction.

Constraints in Patch test

Concentration of ingredient Solid: mild than liquid or semisolid Individual basis: some may produce reaction very soon than others. Seasonal changes: Summer season due to pricks formation around the area of test due to occlusion Volatile subs. may acts as primary irritants

Report of Patch Test

Concentration of chemical / agent, Amount used, Site of application, No. of days patch was left on the skin, Period after the removal of patch when observations were made.

DRAWBACKS OF PROPHETIC TEST

Quick absorption of potential skin irritant thr skin Rapid evaporation of volatile skin irritants from patch Small amount of test sub. is used as compared to large qty. actually used Small area of skin is used as compared to actual S.A. of skin Short exposure of skin to cosmetic than the exposure actual in use.

Various tests to be performed for safety

Cosmetics .potential for adverse reaction Ex.
Skin irritation Allergic sensitization Contact urticaria Stinging Photo toxicity Photo allergy

Skin irritation
Irritation: state of inflammation or painful reaction to allergy or cell-lining damage.
inflammatory responses such as erythema, oedema or vesiculation

Irritants: Substances that damage the skin by toxic reaction

Primary irritants Secondary irritants

How to perform the test?

Tests for cosmetics

Test
1.Skin irritation test 2.Oral toxicity limit test 3.Mucous membrane irritancy test
a. Eye irritancy and mucous membrane irritancy Oral mucous irritancy

Type of product
Skin, hair and lip products Dentrifrices, mouth wash and lip products

Hair care and eye area cosmetics Dentrifrices and mouth wash

b.

Sensitivity testing
Draize patch test Area: hairless, 10 cm2, 2-3 cm apart, 4 patches Applicant: 12 ply surgical gauze on polythene sheet on surgical adhesive plaster

Draize patch test

Quantity: 0.5 ml - liquids 0.5gm - solids/semisolids Positive control: sodium lauryl sulphate (2-20%) Position maintaining: wrapping whole trunk with cloth/canvas bandage Duration of exposure- 24hrs/4hrs Duration of response- 24/48/72 hrs

Draize patch evaluation

Erythema / eschar formation
No erythema Very slight erythema Well defined erythema Moderate to severe erythema Severe erythema and slight eschar formation

Score
0 1 2 3 4

Draize patch evaluation

Oedema formation
No oedema Very slight oedema Slight oedema (defined edges) Moderate oedema(1 mm raising) Severe oedema(1 mm raising beyond area of exposure)

Score
0 1 2 3 4

Oedema: abnormal accumulation of fluid beneath the skin, or in one or more cavities of the body.

Draize patch scoring

Average of 6 animals= a Primary irritancy index= a/12
Index Evaluation

0.00 0.04 0.99 1.00 1.99 2.00 2.99 3.00 5.99 6.00 8.00

No irritation Irritation barely perceptible Slight irritation Mild irritation Moderate irritation Severe irritation

IRRITATION TEST
Indian standards Patch test Guinea pigs/rabbits No. of animals: 10 Patch Area : 2 cm2 Area: without hairs No use of bandage Cosmetic used as such

IRRITATION TEST
Base is painted on skin If no reaction. Test carried out in 10 human volunteer Application on forearm No reactionsafe

REPEATED INSULT TEST

Described by Draize so called Draize test. Apply same conc. found in product in strong base. Strong base may be saline solution to such nonirritating, nonsensitizing organic solvent like dimethyl phthalate.
0.5gm/0.5 ml of sample Site: back/upper arm skin Duration: 24 hours Observation: 15-20 min after patch removal 24 hours rest periodsecond patch different site 10 consecutive exposure10-14 days rest period retest application Comparison of reactions of retest and original application Selection of test subjects 200 test subjects Ideally initially 10 animals10 human volunteer

PHOTOPATCH TEST
Some substances are not harmful by themselves but become harmful when exposed to sun light Wavelength 300- 800 nm Substance applied in duplicate in a manner same for standard patch test After 24 hrs one patch is exposed to sunlight / uv light for 30 min

Exposed patch covered again One adjoining area of skin expose to sunlight in similar manner to patchcontrol site After 24 hrs. examination of patches No reaction no phototoxicity

TEST FOR SENSITIZING POTENTIAL

Standard patch test with same chemical / cosmetic .in 200 volunteers after an interval of 10-14 days Persons showing reactions.sensitizing potential of the substance Provocative patch test Weak sensitizing agents 10-15 applications of a substance on alternate days on same spot in 10 volunteers

TEST FOR SENSITIZING POTENTIAL

Standard patch test performed after 10 days of last application Test will give idea about developing sensitizing potential In case of unknown chemicals, ideally performed on animals like gunia pigs/rabbits

USE TEST
Cosmetic to used and adverse effects are observed No. of applications: 15 No adverse reactions released for trials

RESTRICTED CONSUMER TEST

After passing tests for irritant potential and sensitivity potential Release of 5000-10000 units to market In case of any adverse reactions, further testing with appropriate methods

Stability of cream & lotion

Storage test:
- 200 C for 24 hrs. (thaw) - 50 C for 1 week (thaw), AT in dark place for 4 months, test each month, 400C for 3 months, test each month, 500C for 1 week, test at 1, 3 & 7th day.

Thaw: to pass or change from a frozen to a liquid or semi liquid state

Temperature swing test:

Freeze & thaw cycles during six weeks,
one cycle comprises 24 hrs. storage at -50C 24 hrs. at 400C

Judge & interpret the result

Centrifugation test:
Performed at AT:
Either for 10 mins. At 10,000 rpm or For 30 mins. At 3000 rpm

Specific value is considered and scale the rating as follow:

Stable, Slight inhomogenity, Beginning of separation, Marked separation, Complete separation

Questions ?????
What is the purpose of testing the cosmetic on animal? Enlist various tests to be performed for safety. Write in brief about patch test. Write principle behind irritancy test of cosmetics? Explain in brief irritancy test? Write a note on safety of cosmetic products. Write a note on stability of cosmetic products.