[反相高效液相色谱法（RP-HPLC）分析尿液中的吡啶啉和脱氧吡啶啉]

[Analysis of the pyridinoline and deoxypyridinoline in urine by reversed-phase high performance liquid chromatography (RP-HPLC)].

作者信息

Weng J, Liao Y, Yu B

机构信息

Research Unit of Endocrinology, First Affiliated Hospital, Sun Yat-sen University of Medical Sciences, Guangzhou, 510080.

出版信息

Se Pu. 1997 Nov;15(6):521-3.

PMID:15739338

Abstract

A method for the simultaneous and rapid determination of pyridinoline and deoxypyridinoline in urine using RP-HPLC has been established. After hydrolysis for 18 hours at 107 degrees C in 6 mol/L HCl, the urine sample was prefractionated by partition chromatography using a column packed with cellulose CF1. The appropriate fractions were freeze-dried. The samples were analyzed by HPLC with a 5 microm Spherisorb C18 column, a mobile phase of MeOH/H2O containing 0.1% heptafluorobutyric acid at a rate of 1.2 mL/min and fluorescence detector monitoring at lambda(ex) 290 nm and lambda(em) 400 nm. The limit of a detection for PYD was 10 nmol/L and DPD 7 nmol/L. The recovery of PYD was 91.5% and that of DPD was 106.1%. Within day CV for the two compounds were 1.39% and 0.16% respectively and day-to-day CV 3.71% and 1.32% respectively.

摘要

已建立一种使用反相高效液相色谱法同时快速测定尿液中吡啶啉和脱氧吡啶啉的方法。尿液样品在6mol/L盐酸中于107℃水解18小时后，使用填充有纤维素CF1的柱通过分配色谱法进行预分离。将合适的馏分冷冻干燥。样品通过高效液相色谱法进行分析，使用5μm Spherisorb C18柱，流动相为含有0.1%七氟丁酸的甲醇/水，流速为1.2mL/min，荧光检测器在λ(ex)290nm和λ(em)400nm处进行监测。吡啶啉的检测限为10nmol/L，脱氧吡啶啉的检测限为7nmol/L。吡啶啉的回收率为91.5%，脱氧吡啶啉的回收率为106.1%。这两种化合物日内变异系数分别为1.39%和0.16%，日间变异系数分别为3.71%和1.32%。