一种用于鉴定小组织样本中可还原胶原交联的新快速方法。
A new rapid method for the identification of reducible collagen cross-links in small tissue samples.
作者信息
Smolenski K A, Avery N C, Light N D
出版信息
Biochem J. 1983 Aug 1;213(2):525-32. doi: 10.1042/bj2130525.
A normal-phase high-pressure liquid-chromatography system was used with amino-propyl-bonded silica as the column packing in order to resolve amino acids, reduced amino acid derivatives and reduced cross-links of collagen. The method utilized a solvent system comprising acetonitrile and 10 mM-potassium phosphate buffer, pH 4.3, as described by Schuster [(1980) Anal. Chem. 52, 617-620]. With modifications to the original gradient elution it was possible to resolve fully the radiolabelled components of reduced collagen in one run of less than 80 min. The method is very sensitive, and small biopsy samples can readily be investigated. Although solute retention times gradually decreased with repeated use, little loss of resolution of the reducible cross-links of collagen occurred during a 30-day trial period.
使用正相高压液相色谱系统，以氨丙基键合硅胶作为柱填料，用于分离氨基酸、还原型氨基酸衍生物和胶原蛋白的还原型交联物。该方法采用了由乙腈和10 mM磷酸钾缓冲液（pH 4.3）组成的溶剂系统，如Schuster所述[(1980) Anal. Chem. 52, 617 - 620]。对原始梯度洗脱进行修改后，在不到80分钟的一次运行中就可以完全分离还原型胶原蛋白的放射性标记成分。该方法非常灵敏，小活检样本很容易进行研究。尽管溶质保留时间随着重复使用而逐渐缩短，但在30天的试验期内，胶原蛋白可还原交联物的分离度几乎没有损失。
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