YY 0719.7-2011Ophthalmic optics—Contact lens care products - Part 7: Biological evaluation test methods (English PDF)

3.7 Preparation before test

The cells that have been cultured for 48 h ~ 72 h with good morphology and vigorous growth are made into cell suspension (trypsin, disodium edetate or pancreatin + disodium edetate digestion solution can be used), to perform cell count to determine the cell suspension's concentration; meanwhile prepare a certain concentration of cell suspension according to the requirements of the test method.

3.8 Test method - Agar diffusion method

3.8.2 Add the cell suspension to a phi60 mm culture dish; each culture dish is 4.5 mL, a total of 6 culture dishes.

3.8.3 Put all culture dishes into a 5% CO2 incubator (containing 5% carbon dioxide gas by volume, the same below), to culture it at (37 ± 1) °C saturated humidity for 24 hours; the cells grow into a single layer.

3.8.4 Discard the original culture medium; add 4.5 mL of "agar medium" to each culture dish. Culture in a CO2 incubator for about 30 minutes to make it solidify.

The agar medium is the mixture of double cell culture medium and 0.5% tissue culture agar in equal amounts.

Note: After mixing and adding to the culture dish, it must not be solidified; at the same time, avoid problems such as high temperature denaturation of proteins in the medium or degradation of amino acids.

3.8.5 Add 4.5 mL of newly prepared neutral red active staining solution (or other cell active staining solution) to the surface of the solidified agar; culture in the dark at (37 ± 1) °C for 0.5 h; suck out the excess staining solution.

3.8.6 Use a filter membrane which has a diameter of 0.9 cm to draw 100 µL of the test solution; gently place the filter on the surface of the agar; operate 2 dishes in parallel for each of the positive control group, negative control group and test solution group.

3.8.7 Incubate in a 5% CO2 incubator at (37 ± 1) °C saturated humidity for 24 h ~ 48 h.

a) The name of the test product;

b) Production batch number or production date;

c) Preparation method of test solution;

d) Test medium, cell lines and negative, positive and other control substances;

e) Test procedure;

f) Cell morphology description, measurement data and other conditions.

3.8.1 Prepare a cell suspension of 1 x 10^5 cells/mL

4.1 Purpose

This test evaluates the potential of contact lens care products to produce skin irritation under test conditions.

Note 1: Based on animal welfare requirements, according to GB/T 16886.2, if the test product has skin irritation, the eye irritation test is no longer carried out.

Note 2: If the manufacturer uses the same or similar contact lens care products currently commercially available on the market, the eye irritation test can be performed directly.

4.2 Main equipment and appliances

Animal balance, syringe.

4.3 Preparation of test animals

According to the provisions of GB/T 16886.10.

4.4 Preparation of test solution

4.4.1 For contact lens care products that directly contact human eyes (such as eye wetting solution, multi-functional care solution), directly take the sample as the test solution.

4.4.2 For contact lens care products that indirectly contact the eyes (such as hydrogen peroxide care solution), simulate actual use operations according to the product instructions; take the product that eventually contacts the human body as the test solution.

hydrogen peroxide care solution), simulate actual use operations according to the product instructions; take the product that eventually contacts the human body as the test solution.

4.5 Test procedure and result evaluation

5.4 Test procedure and result evaluation

Operate and evaluate according to the acute exposure test method in GB/T 16886.10.

5.5 Test report

The following information shall be given in the test report:

a) The name of the test sample;

b) Production batch number or production date;

c) Description of the test animal;

d) Preparation of test solution;

e) Test procedure;

f) Observation records;

g) Evaluation of results.

6.1 Purpose

This test is to evaluate the oral toxicity potential of contact lens care products in a single oral dose.

6.2 Main equipment and appliances

Syringe, gavage needle, animal balance.

6.3 Preparation of test animals

6.3.1 The test uses healthy, newborn mice, the same strain and the same source, the female mice are non-pregnant, weighing 17 g ~ 23 g. It allows the mice to adapt to the laboratory environment before the test.

6.3.2 Each group of tested mice uses 10 mice, half male and female. 5 mice of the same sex are kept in each cage and marked.

6.4 Preparation of test solution

The following information shall be given in the test report:

a) The name of the test sample;

b) Production batch number or production date;

c) Description of the test animal;

d) Preparation of test solution;

e) Test procedure;

f) Observation records;

g) Evaluation of results.

7.1 Purpose

In this test, the test solution of the contact lens care product is contacted with the skin of the guinea pig, to evaluate the potential of the contact lens care product to cause skin sensitization under the test conditions.

7.2 Reagents

Sodium chloride injection with a mass concentration of 9 g/L, dinitrochlorobenzene (DNCB), sodium lauryl sulfate, Freund's complete adjuvant.

7.3 Main equipment and appliances

Pressure steam sterilizer, electric razor, syringe, animal balance.

7.4 Preparation of test animals

According to the provisions of GB/T 16886.10.

7.5 Control sample

Sodium chloride injection with a mass concentration of 9 g/L.

7.6 Preparation of test solution

7.6.1 For contact lens care products that directly contact human eyes (such as eye wetting solution, multi-functional care solution), directly take a sample as the test solution. […]