Quantitation of hydroxyproline in bone by gas chromatography-mass spectrometry.
A validated gas chromatography (GC)-mass spectrometric (MS) method for the analysis of hydroxyproline in rat femur is reported. Hydroxyproline in bone hydrolysates was extracted with an anion exchange resin and the N(O)-tert-butyldimethylsilyl derivatives analyzed by GC-MS. The hydroxyproline concentration was estimated relative to pipecolic acid, 3,4-dehydroproline and n-tetracosane as internal standards. The mass-to-charge ratios (m/z) for the ions used for quantitation by single ion monitoring were 314 m/z for hydroxyproline, 198 m/z for pipecolic acid, 256 m/z for dehydroproline and 57 m/z for n-tetracosane. A coefficient of variation of 5.8% was achieved and the limit of detection was calculated to be 0.233 micromol/l bone hydrolysate.
Delport
M
M
Department of Chemical Pathology, University of Pretoria, P.O. Box 2034, Pretoria 0001, South Africa.
Maas
S
S
van der Merwe
S W
SW
Laurens
J B
JB
D000818:Animals; D001842:Bone and Bones / Q000737:chemistry*; D002138:Calibration; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006909:Hydroxyproline / Q000032:analysis*; D051381:Rats; D012680:Sensitivity and Specificity
D006909:Hydroxyproline