Hydroxyproline Levels in Mouse Tissue - Use of Traditional Colorimetric Assay Kit vs. High Resolution LC/MS

摘要

4-Hydroxy-L-proline, or hydroxyproline (HP), a principal and stabilizing component of collagen, is often used as an indicator of collagen turnover or deposition in certain disease states such as fibrosis. HP can be measured using a variety of techniques, including commercially available colorimetric assay kits or by more direct analytical methods such as LC/MS. During development of an in-house in vivo model, significant variability in hydroxyproline levels determined using a purchased colorimetric assay kit was observed. To evaluate if the variability was a result of the detection assay, sample processing (i.e. chemical formation of hydroxyproline during hydrolysis) or inherent variability in the animal model, we compared hydroxyproline levels and variability using both high resolution LC/MS vs. the commercial assay kit.