, is able to neutralize the toxicity of paralytic shellfish poisoning (PSP) toxins. This information offers a groundwork on investigating the detoxifcation of the PSP. Unfortunately, whether or not the treatment methods of the hemolymph will affect the PSP toxicity neutralization is still unclear. In this study, the effects of anticoagulants (EDTA, ), extracting methods (syringe needling extraction and tissue homogenizing extraction) and freezing storage condition (with or without sucrose) on PSP toxicity neutralization of the hemolymph were investigated through the mouse assay. The results showed that the mice intraperitoneal injection with the PSP mixed with EDTA-hemolymph were survival, while the mice’s groups mixed with sodium oxalate, sodium citrate and heparin sodium (28.89 ± 0.59) min and (14.08 ± 0.6) min, which to be 5.4, 5.3 and 2.6 times to that of corresponding control, respectively. Low EDTA concentration was efficacious in anticoagulation of the hemolymph and preserving a good PSP neutralization, which seemed to be repressed with in presence of excessive EDTA. Hemolymph extracted by syringe needle showed higher PSP neutralization than that of extracted by tissue homogenization. While the EDTA concentration was at 5 mmol/L in 20 mmol/L phosphate buffer (pH 8.0), 67% of the mice for the hemolymph group extracted by syringe needle died at more than 30 minutes. By comparison, the mice’s death time for hemolymph group extracted by tissue homogenization ranged at 8.124.0 min. Moreover, sucrose could protect the hemolymph against from freeze denaturation. After freezing for 2 weeks, the PSP neutralization of the hemolymph with sucrose remained unchanged, while that of the hh without sucrose decreased. These results indicate that the PSP neutralization of hemolymph was remarkably effected by anticoagulants, extraction methods and freezing storage condition.