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(10μm,250mm×10mm)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。
A reversed-phase high performance liquid chromatographic-electrospray ionization tandem mass spectrometric method ( RP-HPLC/MS^2 ) was developed for the analysis and identi-fication of a reaction mixture of L-hydroxyproline oligo-peptides. The mixture was separated on a YWG C8 column ( 10 μm, 250 mm × 10 mm) using acetonitrile-watr 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 ob-tained. 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.