Microderivatization of 4-[ 18O]hydroxyproline and quantitation with a benchtop mass spectrometer

• Published in: Journal of chromatography Vol. 562; no. 1; pp. 625 - 637
• Main Authors: Warner, Charles D., Branch, Steven K., Burke, John F., Chambers, Louis M., Young, Vernon R.
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 02.01.1991; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Chromatography, Gas; Collagen - analysis; Fundamental and applied biological sciences. Psychology; Gas Chromatography-Mass Spectrometry; General aspects, investigation methods; Hydroxyproline - analogs & derivatives; Hydroxyproline - analysis; Indicators and Reagents; Mass Spectrometry; Molecular Weight; Oxygen Radioisotopes; Pipecolic Acids - analysis; Proteins; Rats; Regression Analysis; Skin - chemistry
• ISSN: 0378-4347
• DOI: 10.1016/0378-4347(91)80613-H

Accurate estimation of in vivo turnover rates of collagen is complicated by amino acid reutilization. It was previously shown that the ideal, non-recycling tracer was [ 18O]hydroxyproline synthesized in vivo. The analytical method for measuring turnover rates with [ 18O]hydroxyproline must include analyte quantitation for pool size determination and isotope ratio measurement for determining levels of label incorporation. For ease of use and widest availability, a benchtop gas chromatograph—mass spectrometer in the electron-impact ionization mode was chosen. Here we present a versatile procedure for hydroxyproline derivatization that is well suited for routine, large-scale determination of analyte concentrations and relative levels of 18O incorporation.