Determination of hydroxyproline in rat kidney tissue by HPLC with pre-column derivatization / 浙江大学学报·医学版

ABSTRACT

<p><b>OBJECTIVE</b>To establish a pre-column derivatization HPLC method for quantitation of hydroxyproline (Hyp) in rat kidney tissue by reversed-phase assay.</p><p><b>METHODS</b>Rat kidney samples were hydrolyzed in 6 mol x L⁻¹ HCl under 110 degree for 24 h, then 9-fluorenylmethyl chloroformate (FMOC) was added for the pre-column reaction. The HPLC analysis was performed on a Phenomenex C₁₈ column using sodium acetate buffer, methanol and acetonitrile as mobile phase with a flow rate of 1.0 ml min⁻¹ and UV detective wavelength 265 nm at 40 degree.</p><p><b>RESULTS</b>Linear range was 15.30-612.00 mg x L⁻¹ (correlation coefficient was 0.9999). Recovery rate was 97.4%-103.9%.</p><p><b>CONCLUSION</b>The established method is simple, accurate and sensitive to analyze Hyp content in rat kidney tissue.</p>