Colwell A, Russell R G, Eastell R
Department of Human Metabolism and Clinical Biochemistry, University of Sheffield Clinical Sciences Centre, UK.
Eur J Clin Invest. 1993 Jun;23(6):341-9. doi: 10.1111/j.1365-2362.1993.tb02034.x.
The measurement of the 3-OH pyridinium compounds, pyridinoline (Pyr) and deoxypyridinoline (Dpyr), in urine by high performance liquid chromatography is potentially useful in clinical studies, since they are specific biochemical markers of bone resorption. The aims of the present study were to improve assay performance and optimize sample collection. An isocratic high performance liquid chromatogram (HPLC) separation with baseline resolution was accomplished within 4 min using heptafluorobutyric acid as an ion-pair. The sample preparation for HPLC, using CF1 cellulose, produced uncontaminated samples with a recovery higher than 90% for both crosslinks. An elastin-derived material, tentatively identified as isodesmosine (Ides), was also tested and proved to be a suitable internal standard. Use of this standard improved assay precision. The effect of an oral gelatin load on the excretion of Pyr and Dyr was investigated. The creatinine corrected excretion of Pyr and Dpyr was unchanged over a 6 h period, in contrast to the 10-fold increase in the excretion of urinary hydroxyproline with a peak 2-4 h after ingestion. In 20 postmenopausal women, 2 h fasting morning urine results correlated with results from 24-h urine collections Dpyr/Cr (r = 0.70, n = 20). There was a day-to-day variation of 26% in adults studied for 10 days.
通过高效液相色谱法测定尿液中的3-羟基吡啶化合物吡啶啉（Pyr）和脱氧吡啶啉（Dpyr）在临床研究中可能具有重要价值，因为它们是骨吸收的特异性生化标志物。本研究的目的是提高检测性能并优化样本采集。使用七氟丁酸作为离子对，在4分钟内实现了具有基线分辨率的等度高效液相色谱（HPLC）分离。采用CF1纤维素进行HPLC的样本制备，得到的样本无污染，两种交联物的回收率均高于90%。还对一种暂定为异锁链素（Ides）的弹性蛋白衍生物质进行了测试，结果证明它是一种合适的内标。使用该内标提高了检测精度。研究了口服明胶负荷对Pyr和Dyr排泄的影响。与摄入后2 - 4小时达到峰值的尿羟脯氨酸排泄量增加10倍相比，Pyr和Dpyr经肌酐校正后的排泄量在6小时内没有变化。在20名绝经后女性中，空腹晨尿2小时的检测结果与24小时尿液收集的Dpyr/Cr结果相关（r = 0.70，n = 20）。在为期10天的研究中，成年人的每日变化率为26%。
