T/GDCQMA 005-2024 Cosmetic soothing efficacy test - In vitro skin keratinocyte inflammatory factor testing method

Scope

Scope: This document specifies a test method for evaluating the potential soothing effects of cosmetics or their raw materials—the in vitro keratinocyte inflammatory factor test. The test results can serve as one of the supporting evidence for determining whether a cosmetic or its raw materials have soothing effects. This document applies to the testing of samples that achieve soothing effects through inflammatory response pathways induced by external stimuli. Main technical content: 5. Test environment and material requirements 5.1 Test environment: The cell culture cleanroom used in this method shall comply with the requirements of GB/T 42398-2023. The environmental temperature should be controlled at 22°C ± 4°C, and the humidity should be controlled at 45% to 65%. The pressure difference between the clean area and the non-clean area, as well as between clean areas with different air cleanliness levels, should be > 10 Pa. A suitable pressure gradient should be maintained between different functional and grade rooms in the clean area to prevent contamination and cross-contamination. The total air supply volume (non-unidirectional flow) should include 10% to 30% fresh air. The noise level (in quiet state) should be ≤ 65 dB(A). The illumination intensity should be > 300 lx. 5.2 Cell selection: Immortalized human keratinocytes should be selected, preferably from cell strains sourced from ATCC or ECACC. The cells should be free from contamination by external factors such as bacteria, fungi, and mycoplasma. 5.3 Instruments and equipment 5.4 Reagents 5.5 Consumables 6. Test method 6.1 Basic requirements: All solutions, glassware, pipette tips, etc. should be sterile, and all operations should be conducted in a clean environment. 6.2 Cell preparation 6.3 Cell culture conditions 6.4 Test samples 6.4.1 Preparation of test samples 6.5 Determination of test sample concentration 6.6 IL-6 expression detection 6.6.1 Cell processing 6.6.2 ELISA detection of IL-6 expression: Tra

Development Information

Related Standards