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先前的研究工作曾发现,在热适应兔脑室液中有一种能诱使体温降低的活性物质——致热适应因子(HAIF),经Sephadex G25柱层析分离发现存在于P_4组分中。本研究用RP-HPLC进一步从G25-P_4组分中分离出HAIF。活性检测表明,其中保留时间4.61 min的RP-HPLC_3明显降低丘脑下部前区注射兔的肛温。经Waters 990+光电二极管检测的RP-HPLC_3三维图鉴定为色谱纯。RP-HPLC_3的剂量-效应曲线呈高度拟合的抛物线相关;丘脑下部前区注射的适宜剂量相当于3.4～7.2×10~(-10)g HAIF。这些结果证明,RP-HPLC分离得到了高纯度、高效价的HAIF。
Previous studies have found that there is a heat-induced adaptation factor (HAIF), a substance that can induce a decrease in body temperature, in hot adapted rabbit ventricular fluid and is found in the P 4 fraction by Sephadex G25 column chromatography. In this study, RP-HPLC was used to further isolate HAIF from G25-P_4 fraction. The activity assay showed that RP-HPLC_3, with a retention time of 4.61 min, significantly reduced the rectal temperature of rabbit injected into the anterior hypothalamus. The RP-HPLC_3 three-dimensional map of the Waters 990+ photodiode was identified as chromatographic pure. The dose-response curve for RP-HPLC_3 was highly fitted to a parabolic correlation; the appropriate dose for the pre-subthalamic injection corresponded to 3.4 to 7.2 x 10 ~ (-10) g HAIF. These results demonstrate that RP-HPLC separates HAIF with high purity and high titer.