Bünger H, Kaufner L, Pison U
Klinik für Anästhesiologie und operative Intensivmedizin, Medizinische Fakultät Charité, Humboldt-Universität zu Berlin, Germany.
J Chromatogr A. 2000 Feb 18;870(1-2):363-9. doi: 10.1016/s0021-9673(99)01073-0.
A new method for the separation and quantification of two hydrophobic lung surfactant proteins (SPs) is described. It is based on size-exclusion chromatography using the apolar stationary phase butyl silicagel with a pore size of 30 nm and isocratic elution with chloroform, methanol and trifluoroacetic acid. The samples were prepared from sheep lung lavage fluid by centrifugation and fractional extraction with butanol and chloroform-methanol. The chromatograms show three peaks in the elution order SP-B, SP-C and lipids. A small peak ahead of SP-B, which disappeared after reduction with 2-mercaptoethanol, was oligomeric SP-B. The response of the evaporative light-scattering detector was non-linear. For preparative high-performance liquid chromatography ultraviolet detection at 279 nm is recommended.
本文描述了一种分离和定量两种疏水性肺表面活性蛋白（SPs）的新方法。该方法基于尺寸排阻色谱，使用孔径为30 nm的非极性固定相丁基硅胶，并采用氯仿、甲醇和三氟乙酸进行等度洗脱。通过离心以及用丁醇和氯仿 - 甲醇分步萃取，从羊肺灌洗液中制备样品。色谱图显示出按洗脱顺序为SP - B、SP - C和脂质的三个峰。在SP - B之前有一个小峰，用2 - 巯基乙醇还原后消失，该小峰为寡聚体SP - B。蒸发光散射检测器的响应是非线性的。对于制备型高效液相色谱，建议采用279 nm的紫外检测。
