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建立了FMOC柱前衍生液高效液相色谱法测定鱼皮胶原肽中羟脯氨酸含量的分析方法。使用芴甲基氯甲酸酯(FMOC-Cl)作为衍生试剂对鱼皮胶原肽水解液进行衍生,采用Venusil MP C18(4.6 mm×250 mm,5μm)色谱柱,以乙酸盐缓冲液与乙腈为流动相进行梯度洗脱,荧光检测器激发波长250 nm,发射波长395 nm下外标法定量。所测羟脯氨酸的线性关系良好,检出限为0.03μg/m L,平均回收率在97.7%~99.9%之间,相对标准偏差为2.0%(n=6)。
A method for the determination of hydroxyproline in collagen collagen peptide of fish skin by high performance liquid chromatography with precolumn derivatization was established. Derivatization of fish skin collagen peptide hydrolysates was carried out using fluorene methyl chloroformate (FMOC-Cl) as a derivatization reagent, using a Venusil MP C18 (4.6 mm × 250 mm, 5 μm) column with acetate buffer and acetonitrile The mobile phase was eluted with gradient elution. The fluorescence detector was excited at 250 nm and the emission wavelength was 395 nm. The linearity of hydroxyproline was good. The detection limit was 0.03 μg / ml. The average recovery was between 97.7% and 99.9%. The relative standard deviation was 2.0% (n = 6).