A SIMPLE METHOD TO DETERMINE NANOGRAM LEVELS OF 4-HYDROXYPROLINE IN BIOLOGICAL TISSUES
A SIMPLE METHOD TO DETERMINE NANOGRAM LEVELS OF 4-HYDROXYPROLINE IN BIOLOGICAL TISSUES
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DOI：
10.1016/0003-2697(81)90261-x
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发表时间：
1981-01-01
影响因子：
2.9
通讯作者：
HEE, SSQ
中科院分区：
文献类型：
--
作者：
JAMALL, IS;FINELLI, VN;HEE, SSQ
介绍了一种简单实用的组织水解液中纳克级4-羟脯氨酸的定量方法。采用酸水解法从组织匀浆中释放羟脯氨酸。然后用氯胺T氧化游离的羟脯氨酸以产生吡咯型化合物。添加埃利希试剂导致形成最大波长为558 nm的发色团。吡咯和羟脯氨酸标准品和组织水解物的最佳测定条件，特别是氯胺T浓度不同。基于使用吡咯或羟脯氨酸的外标法计算回收率可能会产生相当大的误差。这就需要使用标准加入法。在这些条件下，羟脯氨酸的回收率为97 ± 10%。百分之八水解产物量为25-100 μ l，对应于1.2 - 5 mg组织（湿重），不存在实质性的基质效应。成年大鼠肺组织羟脯氨酸含量始终为3.78 ± 0.001。0.15μ mol/g湿重。
A simple and practical method for the quantitation of nanogram levels of 4-hydroxyproline in tissue hydrolysates is described. Optimum conditions were employed to release the hydroxyproline from tissue homogenates by acid hydrolysis. The free hydroxyproline was then oxidized by chloramine T to produce a pyrrole-type compound. The addition of Ehrlich''s reagent resulted in the formation of a chromophore with a wavelength maximum at 558 nm. The optimum conditions for the assay, notably the chloramine T concentration, were different for pyrrole and hydroxyproline standards, and tissue hydrolysates. The calculation of recoveries based on the external standards method using either pyrrole or hydroxyproline could be subject to considerable error. This necessitated the use of the method of standard additions. The recovery of hydroxyproline under these conditions was 97 .+-. 8%. No substantial matrix effects exists with hydrolysate quantities ranging from 25-100 .mu.l corresponding to 1.2 to 5 mg of tissue (wet wt). The hydroxyproline content of adult rat lung was consistently 3.78 .+-. 0.15 .mu.mol/g wet wt.