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[Determination of hydroxyproline in rat kidney tissue by HPLC with pre-column derivatization]
Er-gang Liu1, Xu-chun Fu, Ying Qian
1Institute of Materia Medica, Zhejiang University City College, Hangzhou 310015, China.
Objective:
To establish a pre-column derivatization HPLC method for quantitation of hydroxyproline (Hyp) in rat kidney tissue by reversed-phase assay.
Methods:
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.
Results:
Linear range was 15.30-612.00 mg x L⁻¹ (correlation coefficient was 0.9999). Recovery rate was 97.4%-103.9%.
Conclusion:
The established method is simple, accurate and sensitive to analyze Hyp content in rat kidney tissue.
