摘要
我们对影响羟脯氨酸（ＨＹＰ）和羟赖氨酸（ＨＹＬ）分析的柱床高度、柱温和洗脱条件进行探讨，在氨基酸分析仪建立了胶原特异性氨基酸的快速测定方法，本方法简单、快速（每份样品分析时间仅１８ｍｉｎ）、回收率高（＜９５％），重复性好（批间ＣＶ＜６％，批内ＣＶ＜４％）、实用性强（可测定胶原水解液及生理体液中的ＨＹＰ和ＨＹＬ），分析效率是常规方法的１２倍，是胶原特异性氨基酸测定、胶原分型及胶原含量估算的理想方法。用本方法对兔正常软骨和８例硬皮病患者皮肤的测定结果表明：透明软骨和纤维软骨的ＨＹＰ和ＨＹＬ含量、胶原类型及胶原总量不同（Ｐ＜０．０１），硬皮病皮肤中的ＨＹＬ和ＨＹＬ明显高于正常人皮肤（Ｐ＜０．０１）。
s:We improved the column heigth, column temperature and elution buffer, which affect the determination of hydroxyproline(HYP) and hydroxylysine(HYL). We set up a fast method of determination collagen specific amino acids using amino acid analyzer.Our method is simple, fast(only using 18 minutes) and has high recovery(>950%), good repeatability(CV<60% among groups, CV< 4% within group) and high efficiency(as twelve times as nornal method). It can be used in both collagen hydrolyzate and physiological fluid. It is an ideal method of determination collagen specific amino acids, classfying and estimating quantity of collagen. We determinated the normal rabbits cartilage and sclerodernia patients skin using our method The results indicated: hyaline cartilage and ribrocartilage are different from the quantity of HYP and HYL, from kinds and total quantity of collagen(P<0.01); HYP and HYL in scleroderma patients' skin are much higher than normal people(P<0.01).
关键词
胶原
羟脯氨酸
羟赖氨酸
氨基酸
分析
collagen
hydroxyproline
hydroxylysine
amino acid analyzer
fast determination
