利用RP-HPLC分析柑橘果肉水溶性蛋白质的差异

RP-HPLC Analysis on Water-Soluble Proteins Extracted from Citrus Pulps of Different Genotypes

本文探索了利用超声波细胞破碎仪提取柑橘果肉中水溶性蛋白质的方法,并对流动相及波长等色谱条件进行优化,建立了反相高效液相色谱分离柑橘果肉水溶性蛋白质的技术。优化后的色谱条件为:流动相A为0.05%三氟乙酸甲醇溶液,B为0.05%三氟乙酸水溶液,A液100%～72%,梯度洗脱45 min,再用A液平衡10min,流速为1.0 mL/min,进样量为20μL,检测波长为270 nm。从7个柑橘品种的果肉中共分离出20个不同的蛋白质峰,图谱中各品种的特有蛋白质反映了不同品种不同表型性状的重要信息。

Protein profile reflects important information regarding phenotype of plants.In this experiment,ultrasonic extraction method was optimized for water-soluble protein extraction from citrus pulp of different genotypes and the proteins were separated by Reverse Phase-High Performance Liquid Chromatograph（RP-HPLC）,and protein profile was obtained and compared.For water-soluble protein analysis,HPLC conditions was optimized as： 0.05% TFA in methanol as mobile A,0.05% TFA in water as mobile B,the injection volume was 20 μL,and the detection was carried out at 270 nm and using a 45-min linear gradient： from 100% to 72% A with the flow rate set at 1.0 mL/min then balance the chromatograph with mobile phase A for 10 min.With the just-mentioned method,a total of 20 proteins were eluted,which might provide important information related with their specific phenotypes.