Clinical Chemistry
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Clinical Chemistry 54: 1546-1553, 2008. First published July 24, 2008; 10.1373/clinchem.2008.105262
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(Clinical Chemistry. 2008;54:1546-1553.)
© 2008 American Association for Clinical Chemistry, Inc.

Endocrinology and Metabolism

Automated HPLC Assay for Urinary Collagen Cross-links: Effect of Age, Menopause, and Metabolic Bone Diseases

Marius E. Kraenzlin1,2,a, Claude A. Kraenzlin2, Christian Meier1,2, Cecilia Giunta3 and Beat Steinmann3

1 Division of Endocrinology, Diabetes, and Clinical Nutrition, University Hospital, CH-4031 Basel, Switzerland; 2 Endocrine Practice and Laboratory, Missionsstrasse 24, CH-4055 Basel, Switzerland; 3 Division of Metabolism & Molecular Pediatrics, University Children’s Hospital, CH-8032 Zurich, Switzerland.

aAddress correspondence to this author at: Missionsstrasse 24, CH-4055 Basel / Switzerland. Fax +41 61 264 97 96; e-mail marius.kraenzlin{at}unibas.ch.

Background: The pyridinium cross-links pyridinoline (PYD) and deoxypyridinoline (DPD) are established markers of bone resorption. We evaluated the analytical and clinical performance of a commercially available PYD HPLC assay and established reference intervals in children and adults.

Methods: We used a commercially available reagent set (Chromsystems Instruments & Chemicals) to measure PYD and DPD in 319 healthy controls (156 premenopausal women, 80 healthy men, and 83 healthy children age 1 month to 14 years) and 397 patients with metabolic bone diseases (postmenopausal osteoporosis, n = 175; male osteoporosis, n = 176; hyperparathyroidism, n = 17; hyperthyroidism, n = 19; Paget disease, n = 10).

Results: The mean intraassay and interassay CVs were <6% and <8% for both PYD and DPD, respectively. The reference interval was constant for premenopausal women in the age group 20–49 years. In men, cross-link values peaked at 20–29 years and decreased thereafter. Women with postmenopausal osteoporosis had significantly higher PYD (51%) and DPD (58%) values compared to premenopausal women. Similar results were found in osteoporotic men. In children the highest values were found in the first weeks and months after birth, followed by a decrease of 50%–60% at age 11–14 years. In metabolic bone diseases cross-link concentrations were significantly increased. The DPD:PYD ratio (mean value approximately 0.2) was remarkably constant in all populations evaluated.

Conclusions: The automated HPLC assay is a precise and convenient method for PYD and DPD measurement. We established reference intervals for adult women and men and for children up to 14 years old. The cross-link concentrations we determined by use of this HPLC method confirm its clinical value in enabling identification of increased bone resorption in patients with metabolic bone diseases.






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