Determination of hydroxyproline in rat kidney tissue by HPLC with pre-column derivatization.
- Author:
Er-gang LIU
1
;
Xu-chun FU
;
Ying QIAN
;
Hai-bo BAI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Chromatography, High Pressure Liquid; methods; Hydroxyproline; analysis; In Vitro Techniques; Kidney; chemistry; Male; Rats
- From: Journal of Zhejiang University. Medical sciences 2012;41(2):188-191
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a pre-column derivatization HPLC method for quantitation of hydroxyproline (Hyp) in rat kidney tissue by reversed-phase assay.
METHODSRat 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.
RESULTSLinear range was 15.30-612.00 mg x L⁻¹ (correlation coefficient was 0.9999). Recovery rate was 97.4%-103.9%.
CONCLUSIONThe established method is simple, accurate and sensitive to analyze Hyp content in rat kidney tissue.
