The aim of this study is to validate an HPLC-fluorescence method for the quantification of free pyridinoline (Pyd), free deoxypyridinoline (D-Pyd), indices of bone degradation, together with galactosyl (Gal-Pyd) and glucosyl-galactosyl (Glu-Gal-Pyd) pyridinolines in non-hydrolyzed healthy women urine (n=7, age=34±7). For the first time, a synthesized D-Pyd superior homologue is proposed as internal standard. For all these compounds, the method is linear in the range 0-170 pmol injected and no matrix effect is found. The coefficients of variation for intra- and inter-day repeatability are between 3.1 and 10.4%. The limit of detection is around 2.3 pmol injected. Free D-Pyd, free Pyd and Glu-Gal-Pyd evaluated in urine account for 40.5±4.7, 215.1±25.0 and 96.4±20.1 pmol/mL (mean±SD), respectively. Gal-Pyd is absent in control urines or under our detection limit. The addition of the new proposed internal standard to urine before any pre-analytical step, allows the unequivocal quantification of the selected bone resorption indices. Moreover, the correct HPLC identification of all selected analytes is guaranteed by the availability of pure corresponding synthesized standards.

Quantification of bone turnover markers in control human urine by HPLC-fluorescence using a new proposed internal standard / E. Monticelli, C.S. Aman, L. Costa, P. Rota, P. Allevi, G. Cighetti, M. Anastasia. ((Intervento presentato al 54. convegno National Meeting of the Italian Society of Biochemistry and Molecular Biology-SIB tenutosi a Catania nel 2009.

Quantification of bone turnover markers in control human urine by HPLC-fluorescence using a new proposed internal standard

E. Monticelli
Primo
;
C.S. Aman
Secondo
;
L. Costa;P. Rota;P. Allevi;G. Cighetti
Penultimo
;
M. Anastasia
Ultimo
2009

Abstract

The aim of this study is to validate an HPLC-fluorescence method for the quantification of free pyridinoline (Pyd), free deoxypyridinoline (D-Pyd), indices of bone degradation, together with galactosyl (Gal-Pyd) and glucosyl-galactosyl (Glu-Gal-Pyd) pyridinolines in non-hydrolyzed healthy women urine (n=7, age=34±7). For the first time, a synthesized D-Pyd superior homologue is proposed as internal standard. For all these compounds, the method is linear in the range 0-170 pmol injected and no matrix effect is found. The coefficients of variation for intra- and inter-day repeatability are between 3.1 and 10.4%. The limit of detection is around 2.3 pmol injected. Free D-Pyd, free Pyd and Glu-Gal-Pyd evaluated in urine account for 40.5±4.7, 215.1±25.0 and 96.4±20.1 pmol/mL (mean±SD), respectively. Gal-Pyd is absent in control urines or under our detection limit. The addition of the new proposed internal standard to urine before any pre-analytical step, allows the unequivocal quantification of the selected bone resorption indices. Moreover, the correct HPLC identification of all selected analytes is guaranteed by the availability of pure corresponding synthesized standards.
23-set-2009
pyridinium crosslink ; glycosilated pyridinoline ; HPLC
Settore BIO/10 - Biochimica
Italian Society of Biochemistry and Molecular Biology-SIB
Quantification of bone turnover markers in control human urine by HPLC-fluorescence using a new proposed internal standard / E. Monticelli, C.S. Aman, L. Costa, P. Rota, P. Allevi, G. Cighetti, M. Anastasia. ((Intervento presentato al 54. convegno National Meeting of the Italian Society of Biochemistry and Molecular Biology-SIB tenutosi a Catania nel 2009.
Conference Object
File in questo prodotto:
Non ci sono file associati a questo prodotto.
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/73198
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? ND
social impact