Assay for urinary desmosines in a healthy pre-pubertal population using an improved extraction technique.
BACKGROUND
Current evidence indicates that increased desmosine excretion reflects the active inflammatory status of some connective tissue diseases. Our goal was to establish a reliable method of detection and to investigate the normal distribution of urinary desmosine excretion in a healthy pre-pubertal population.


METHODS
Urine was collected from healthy volunteers aged four weeks to 12 years old. We modified a published high-performance liquid chromatography (HPLC) method by (a) increasing hydrolysis time and temperature and (b) increasing cellulose column size.


RESULTS
Our modified method had small inter- and intra-assay variability, with coefficients of variation of <6.4% and 5.3%, respectively. There was positive correlation between isodesmosine and desmosine (r(2) = 0.91). There was no significant diurnal or day-to-day variability in total desmosine levels. A reference range for healthy pre-pubertal children aged four weeks to 12 years was established.


CONCLUSIONS
The modified HPLC method is reliable with low variability. The technique can now be applied as a non-invasive research or diagnostic tool for children with chronic lung disease.
Institute for Child Health Research, University of Western Australia.
Kaye R
KR
Winfield
Samantha
S
Gard
G Neil
GN
Kent
Peter D
PD
Sly
Siobhain
S
Brennan
D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003895:Desmosine / Q000592:standards* / Q000652:urine*; D005260:Female; D006801:Humans; D006868:Hydrolysis; D007223:Infant; D007231:Infant, Newborn
D003895:Desmosine