OBJECTIVE： To establish the detection method for pyrogen-related factor interleukin 1β （IL-1β） through optimizing detection condition， and to conduct preliminary methodology study. METHODS： The in vitro fresh human whole blood detection method was used. The bacterial endotoxin standard solution （5， 2， 0.8， 0.32 EU/ml） were added into diluted blood； using diluted RPMI 1640 as blank control， the content of IL-1β in blood sample was determined by ELISA after incubation. The relationship of the addition of different attenuants （RPMI 1640 culture， sterilized normal saline） and fetal bovine serum， final dilution volume fraction （40％， 20％， 10％ and 8.3％） and storage duration （2， 5， 6， 8， 26 h） with the contents of endotoxin IL-1β were investigated， and related coefficient and detection limits were calculated. Different dilution times of Qingkailing injection and Ginaton injection samples and interference solutions were added into diluted blood to detect their recovery. RESULTS： The results indicated that RPMI 1640 media and 40％ diluted blood was more sensitive （detection limit was 0.128 EU/ml， r＝0.993）； while the addition of fetal bovine serum didn’t influence the results. The detection limits of blood sample storied at 4 ℃ for 26 h were 0.128 EU/ml （r＞0.990）. When Qingkailing injection and Ginaton injection were diluted 10， 32 and more times， the detection method was not interfered and the recovery ranged 68％-118％. CONCLUSIONS： Established in vitro fresh human whole blood detection method can be used for the detection of pyrogen， and provides trial evidence for the pyrogen detection of TCM injection.