A rapid and simple method for quantitation of urinary hydroxylysyl glycosides, indicators of collagen turnover, using liquid chromatography/tandem mass spectrometry.
Some glycosides of hydroxylysine, viz., alpha-1, 2-glucosylgalactosyl-O-hydroxylysine and beta-1-galactosyl-O-hydroxylysine, appear to be good indicators of collagen turnover. A simple liquid chromatography/tandem mass spectrometry (LC/MS/MS) method is proposed for measuring these analytes in urine, with no sample preparation except for a dilution step. Quantitation is performed using external calibration with no internal standard. A preliminary survey indicates good intra- and inter-day reproducibility (better than 5 and 8%, respectively). With the present method, the estimated limits of detection (S/N > 3) in urine are 0.8 and 0.5 microM/L for beta-1-galactosyl-O-hydroxylysine and alpha-1,2-glucosylgalactosyl-O-hydroxylysine, respectively. The method is proposed as a robust tool for a large-scale research investigation on collagen turnover.
Casetta
B
B
Applied Biosystems, Monza, Milano, Italy. bruno_casetta@eur.appliedbiosystems.com
Romanello
M
M
Moro
L
L
D002853:Chromatography, Liquid / Q000379:methods*; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D006901:Hydroxylysine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000652:urine; D013058:Mass Spectrometry / Q000379:methods*
D006901:Hydroxylysine; C001295:glucosylgalactosylhydroxylysine; C007817:galactosylhydroxylysine; D003094:Collagen