Abstract: OBJECTIVE To promote the understanding of users of the Chinese Pharmacopoeia on the supplementary methods of the new endotoxin test method and ensure the implementation of the Chinese Pharmacopoeia 2020 edition, introduced the relevant content of the drafting background, revision and application of the recombinant factor C method and the micro-gel method. METHODS Combined the 9251 bacterial endotoxin test application guidelines principle and the research data of recombinant factor C and microgel method, the two methods were clarified. RESULTS Combined with the endotoxin inspection method and current status of horseshoe crab resources, classification of supplementary methods. The introduction, application status and use of recombinant factor C method and microgel method were discussed, and the two supplementary methods were introduced in detail. CONCLUSION The use of recombinant factor C method and microgel method in accordance with the requirements of the Chinese Pharmacopoeia can alleviate the problems caused by the shortage of horseshoe crab resources and ensure the quality and safety of injections.