Microderivatization of 4-[18O]hydroxyproline and quantitation with a benchtop mass spectrometer.
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.
Warner
C D
CD
Shriners Burns Institute, Boston, MA 02114.
Branch
S K
SK
Burke
J F
JF
Chambers
L M
LM
Young
V R
VR
D000818:Animals; D002849:Chromatography, Gas; D003094:Collagen / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry; D006909:Hydroxyproline / Q000031:analogs & derivatives / Q000032:analysis*; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D008970:Molecular Weight; D010104:Oxygen Radioisotopes; D010875:Pipecolic Acids / Q000032:analysis; D051381:Rats; D012044:Regression Analysis; D012867:Skin / Q000737:chemistry
D007202:Indicators and Reagents; D010104:Oxygen Radioisotopes; D010875:Pipecolic Acids; D003094:Collagen; D006909:Hydroxyproline