Abstract: Tetrodotoxin is a widely distributed neurogenic toxin with severe toxicity which can lead to nerve paralysis and cause death by blocking sodium channels and nerve conduction. As one of the most critical risks that threaten the safety of aquatic products, it is strictly controlled by international community. Meanwhile, tetrodotoxin can be used in clinical practice for sedation, analgesia and anesthesia, and has comprehensive application foreground. There are three main methods to detect tetrodotoxin (bioassay, immunoassay and instrumental method). However, it is difficult to produce high-purity tetrodotoxin in enormous quantity because of lack of purification and preparation technology with high separation efficiency and high yield. The paper reviews the research progress in the determination and preparation of tetrodotoxin method, and summerizes the applicability and problem of each method, which provides references for further study.