生物体液中锁链素和异锁链素的富集与分析。

Enrichment and analysis of desmosine and isodesmosine in biological fluids.

作者信息

Cumiskey W R, Pagani E D, Bode D C

机构信息

Sterling Winthrop Pharmaceuticals Research Division, Department of Pharmacology, Collegeville, PA, USA.

出版信息

J Chromatogr B Biomed Appl. 1995 Jun 23;668(2):199-207. doi: 10.1016/0378-4347(95)00092-w.

DOI:10.1016/0378-4347(95)00092-w

PMID:7581855

Abstract

A method has been developed for the enrichment and analysis of the elastin crosslinks, desmosine and isodesmosine, in biological fluids and tissues. It is adapted from published methods, offering improved recovery, sensitivity, resolution, and speed of analysis. Samples were hydrolyzed in 6 M HCl, after which the desmosines were enriched by CF1 cellulose chromatography and analyzed by HPLC with a C18 column. Isodesmosine and desmosine were quantitated based on absorbance at 275 nm, with a limit of detection of approximately 30 pmol and recovery of approximately 66% in urine. Their tR values on our HPLC system were approximately 9 and 12 min, respectively. This method was used to evaluate the daily and weekly variation in the concentrations of desmosine and isodesmosine in human urine. The results suggest that this method can be used to process large numbers of biological samples for analysis of desmosine and isodesmosine.

摘要

已开发出一种用于富集和分析生物体液及组织中弹性蛋白交联物、锁链素和异锁链素的方法。该方法改编自已发表的方法，具有更高的回收率、灵敏度、分辨率和分析速度。样品在6 M盐酸中水解，然后通过CF1纤维素色谱法富集锁链素，并使用C18柱通过高效液相色谱法进行分析。异锁链素和锁链素基于275 nm处的吸光度进行定量，检测限约为30 pmol，尿液中的回收率约为66%。它们在我们的高效液相色谱系统上的保留时间分别约为9分钟和12分钟。该方法用于评估人尿中锁链素和异锁链素浓度的每日和每周变化。结果表明，该方法可用于处理大量生物样品以分析锁链素和异锁链素。