摘要
目的:建立一种测定大鼠肾组织中羟脯氨酸(Hyp)含量的柱前衍生HPLC法。方法:将大鼠肾组织以6 mol·L-1盐酸于110℃水解24 h,加入9-芴甲氧基羰酰氯(FMOC)衍生后,进行HPLC分析。HPLC分析条件为:Phenomenex C18色谱柱(4.60 mm×150 mm,5(m);流动相:醋酸钠缓冲液(pH3.78)-甲醇-乙腈(梯度洗脱);流速:1.0 ml·min-1;柱温:40℃;检测波长:265 nm。结果:羟脯氨酸在浓度范围为15.30～612.00 mg.L-1内线性良好(r=0.9999);回收率为97.4%～103.9%。结论:该方法简单、准确、灵敏,可用于测定大鼠肾组织中羟脯氨酸的含量。
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·L-1 HCl under 110 ℃ for 24 h,then 9-fluorenylmethylchloroformate（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℃.Results： Linear range was 15.30-612.00 mg·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.
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2012年第2期188-191,共4页
Journal of Zhejiang University(Medical Sciences)
关键词
色谱法
高压液相
肾/代谢
羟脯氨酸/分析
离心法
梯密度
柱前衍生
肾组织
Chromatography
high pressure liquid
Kidney/metab
Hydroxyproline/anal
Centrifugation
density gradient
Pre-column derivatization
Kidney tissue
