GB/T 43450-2023 (English Version)

Standard No.: GB/T 43450-2023
Language: Chinese, Available in English version
Release Date: 2023
Published By: General Administration of Quality Supervision, Inspection and Quarantine of the People‘s Republic of China
Latest: GB/T 43450-2023

Scope: This document specifies the method principle, instruments and reagents, test materials, test and result judgment of acute eye irritation test of chemicals. This document is applicable to in vitro detection of acute eye irritation of chemicals. Chemicals are soluble in water and the pH of the test solution should be in the range of 6.5 to 7.8.

Introduction

Interpretation of the standard for in vitro cell test for acute eye irritation of chemicals TRPV1 activity detection method

Overview of the standard

This standard GB/T 43450-2023 "In vitro cell test for acute eye irritation of chemicals TRPV1 activity detection method" was issued by the State Administration for Market Regulation and the National Standardization Administration, and is applicable to the in vitro detection of acute eye irritation of chemicals. This method is based on the activation effect of TRPV1 channels and evaluates the eye irritation of chemicals by detecting changes in calcium ion concentration.

Background of standard formulation

With the widespread application of chemicals, the issue of eye irritation has attracted much attention. Traditional animal experiments have ethical and cost issues, and in vitro alternative methods have become a trend. This standard combines the characteristics of TRPV1 channels and proposes an efficient and reliable in vitro detection method.

Technical Route Analysis

The technical route of this standard includes the following key steps:

Cell culture and preparation
Dye incubation
Sample preparation and testing
Data acquisition and analysis

Technical dimensions	Standard requirements	Implementation points
Cell line selection	Using TRPV1-HEK293 cell line	Ensure that cells are free of contamination and control the number of passages within 25 generations
Reagents and consumables	Including DMEM culture medium, calcium ion dye, etc.	Reagents must meet laboratory water standards and strictly follow aseptic operations
Test equipment	High-throughput real-time fluorescence detection analyzer, etc.	Ensure equipment calibration and accurate data collection

Result determination and application

According to the standard (7.2-7.5), the acute eye irritation of chemicals is determined by the following indicators:

E_c-: The test sample has no stimulating effect of pepper flat hole
E_c+: antagonistic effect of the test sample plus pepper flat hole
AUC value: area under the time-fluorescence intensity curve

Implementation suggestions

To ensure the effective implementation of the standard, it is recommended to take the following measures:

Strengthen laboratory staff training and familiarize themselves with the principle of TRPV1 channel detection
Establish a quality control system to ensure the accuracy of reagents and equipment
Regularly verify cell line activity to avoid contamination
Combine other standards (such as GB/T 21609) for multi-dimensional evaluation

GB/T 43450-2023 Referenced Document

GB/T 21609 Chemicals.Test method of acute dermal irritation/corrosion
GB/T 6682 Water for analytical laboratory use.Specification and test methods
SN/T 2285 Good laboratory practice for cosmetics alternative testing in vitro
SN/T 3899 Guidance on good cell culture practice and sample preparation for cosmetics alternative testing in vitro

GB/T 43450-2023 history

2023 GB/T 43450-2023 Chemical acute eye irritation in vitro cell test TRPV1 activity detection method