L-羟脯氨酸寡肽混合物的高效液相色谱分离与质谱分析

研究了三氯氧磷辅助下L-羟脯氨酸的成肽反应,建立了采用反相高效液相色谱-质谱/质谱联用技术分离鉴定羟脯氨酸寡肽混合物的方法,优化了L-羟脯氨酸寡肽混合物的色谱分离条件。实验以YWG C8柱(10 μm,250 mm×10 mm)为分离柱,以乙腈-0.06%三氟乙酸水溶液(体积比为2∶98)为流动相进行等度洗脱,在正离子模式下对洗脱物进行了电喷雾电离串联质谱鉴定。结果显示,分离出的各组分分别为L-羟脯氨酸二肽、L-羟脯氨酸环二肽和L-羟脯氨酸三肽。

Isolation and Identification of the Mixture of L-Hydroxyproline Oligo-Peptides by Reversed-Phase High Performance Liquid Chromatography and Electrospray Ionization Tandem Mass Spectrometry

A reversed-phase high performance liquid chromatographic-electrospray ionization tandem mass spectrometric method (RP-HPLC/MS2) was developed for the analysis and identification of a reaction mixture of L-hydroxyproline oligo-peptides. The mixture was separated on a YWG C8 column (10 microm, 250 mm x 10 mm) using acetonitrile-water containing 0.06% trifluoroacetic acid (2:98, v/v) as the mobile phase. The separation conditions were optimized. The mixture was analysed with electrospray ionization mass spectrometry (ESI-MS) and ESI-MS/MS in positive mode. MS and MS2 spectra of individual chromatographic peak were obtained. It indicated that there were hydroxyproline dipeptide, hydroxyproline tripeptide and hydroxyproline cyclo-dipeptide in the mixtures, besides hydroxyproline. The described method can be effectively applied to perform identification of hydroxyproline oligo-peptide mixtures.