反相高效液相色谱法测定尿中吡啶醚和脱氧吡啶醚

 尿中吡啶醚（ｐｙｒｉｄｉｎｏｌｉｎｅ，ＰＹＤ）和脱氧吡啶醚（ｄｅｏｘｙｐｙｒｉｄｉｎｏｌｉｎｅ，ＤＰＤ）是骨代谢特异的生化指标。应用高效液相色谱法（ＨＰＬＣ）建立了尿中ＰＹＤ和ＤＰＤ的测定方法。尿液用６ｍｏｌ／ＬＨＣｌ水解后，以纤维素ＣＦ１小柱提取，然后用ＨＰＬＣ测定；色谱系统为ＳｐｈｅｒｉｓｏｒｂＣ１８反相色谱柱，流动相组成为１５％甲醇添加０．１％七氟丁酸，流速为１．２ｍＬ／ｍｉｎ。系统的检测限：ＰＹＤ为１０ｎｍｏｌ／Ｌ，ＤＰＤ为７ｎｍｏｌ／Ｌ；回收率：ＰＹＤ为９１．５％，ＤＰＤ为１０６．１％；日内变异

Analysis of the Pyridinoline and Deoxypyridinoline in Urine by Reversed Phase High Performance Liquid Chromatography (RP HPLC)

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.