A novel method for evaluating pseudoallergy based on β‐hexosaminidase and its application for traditional Chinese medicine injections

Pseudoallergy is a typical and common adverse drug reaction to injections, especially in traditional Chinese medicine injections (TCMIs). At present, the evaluation methods for pseudoallergy include cell methods in vitro and animal methods in vivo. The mast cell evaluation method based on the β‐hexosaminidase (β‐Hex)‐catalyzed substrate, 4‐nitrophenyl‐β‐N‐acetyl‐D‐glucosaminide (4‐NPG), is an important method for the evaluation of drug‐induced pseudoallergy, but it is prone to false positive results and has insufficient sensitivity. In this study, a novel β‐Hex evaluation system with rat basophilic leukemia‐2H3 cells based on high‐performance liquid chromatography‐fluorescence detection (HPLC‐FLD) was established, which effectively increased the sensitivity and avoided false positive results. Cell viabilities were measured by 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl tetrazolium bromide assay. In addition, a method for the determination of histamine, which is another indicator in the development of pseudoallergy, was established to validate the above method. The results of this novel method indicated that two TCMIs (Shuxuening injection and Shenqi Fuzheng injection), which were considered to be pseudoallergenic using 4‐NPG, were not pseudoallergenic. Overall, the novel β‐Hex/HPLC‐FLD evaluation system using Rat basophilic leukemia‐2H3 cells established was effective and precise. It could be used for the evaluation of pseudoallergic reactions caused by TCMIs and other injections.