Determination of hydroxyproline by high pressure liquid chromatography

✍ Scribed by Graham D. Green; Kathleen Reagan

• Publisher: Elsevier Science
• Year: 1992
• Tongue: English
• Weight: 476 KB
• Volume: 201
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(92)90337-7

✦ Synopsis

A rapid, precise, and simple HPLC method provides an assay of hydroxyproline from tissue extracts or solutions of collagen. Samples are hydrolyzed with 6 N HCl, derivatized with phenyl isothiocyanate, and chromatographed on a small, C18 reverse-phase HPLC column. Hydroxyproline (Hyp) is separated from other amino acids and detected by absorption at 254 nm. The method detects 0.40 to 36 micrograms of Hyp with a linear response. Separation requires a total of 6 min, including column cleanup and reequilibration. All components are commercially available, making this a convenient method for routine measurement of collagen concentration.