Yamada T, Pettit G W
Circ Shock. 1977;4(4):379-86.
A simplified paper chromatographic technique for assaying myocardial depressant factor (MDF) is presented. In addition to a serine standard, deproteinized plasma samples from rabbits subjected to hemorrhagic shock were applied to paper and chromatographed. The spot in the chromatograms occurring at a migration distance of 1.6 to 1.8 times that of the standard (RS = 1.6-1.8) was found by bioassay to contain the MDF. The chromatographic spots were developed with ninhydrin and eluted with 3 ml of 1% NaHCO3. The absorbance at 570 nm (A570) of each eluate was measured and compared against the A570 value of the eluted serine standard. A direct linear relationship was shown between MDF chromatographic and bioassay values for each animal. This technique for paper chromatographic assay of MDF is as sensitive, specific, and reliable as bioassay, but offers clear advantages of small sample volume requirement, reduced analysis time, and ease of performance with routinely available laboratory equipment.
本文介绍了一种用于测定心肌抑制因子（MDF）的简化纸色谱技术。除了丝氨酸标准品外，还将遭受失血性休克的兔子的去蛋白血浆样本点样于滤纸上并进行色谱分析。通过生物测定发现，色谱图中迁移距离为标准品1.6至1.8倍（Rf = 1.6 - 1.8）处的斑点含有MDF。用茚三酮显色色谱斑点，并用3 ml 1% NaHCO₃洗脱。测量每个洗脱液在570 nm处的吸光度（A₅₇₀），并与洗脱的丝氨酸标准品的A₅₇₀值进行比较。结果表明，每只动物的MDF色谱值与生物测定值之间呈直接线性关系。这种用于MDF纸色谱测定的技术与生物测定一样灵敏、特异且可靠，但具有明显优势，即所需样本量小、分析时间缩短，并且使用常规实验室设备即可轻松操作。
