Simple and sensitive method for quantification of fluorescent enzymatic mature and senescent crosslinks of collagen in bone hydrolysate using single-column high performance liquid chromatography.
A rapid high performance liquid chromatographic method was developed including an internal standard for the measurement of mature and senescent crosslinks concentration in non-demineralized bone hydrolysates. To avoid the demineralization which is a tedious step, we developed a method based on the use of a solid-phase extraction procedure to clean-up the samples. It resulted in sensitive and accurate measurements: the detection limits as low as 0.2 pmol for the pyridimium crosslinks and 0.02 pmol for the pentosidine. The inter- and intra-assay coefficients of variation were as low as 5% and 2%, respectively, for all crosslinks.
Viguet-Carrin
S
S
INSERM Research Unit 831, Claude Bernard University of Lyon, Lyon, France.
Gineyts
E
E
Bertholon
C
C
Delmas
P D
PD
D000818:Animals; D001842:Bone and Bones / Q000737:chemistry*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003094:Collagen / Q000737:chemistry*; D005453:Fluorescence; D006868:Hydrolysis; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D003094:Collagen