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用与反相高效液相色谱偶联的串联质谱 (RPHPLC MS MS)分析一个固相化学合成的九肽H2 N Tyr Val Asn Val Asn Met Gly Leu Lys CONH2 及其 2个主要的副产物 .根据Fmoc化学 ,在Fmoc PAL PEG·PS树脂上从C端至N端手工操作逐步偶联合成九肽 .偶联完成后 ,用试剂R(90 %TFA ,5 %苯甲硫醚 ,3%二巯基乙烷 ,2 %苯甲醚 )室温 (2 0～ 2 5℃ )处理 2 5h ,进行多肽的去保护和切除树脂 .RP HPLC分析结果表明 ,合成粗品中含有 1个主要成分、2个次要成分及多个微量成分 .用RPHPLC MS MS分别对主要成分和 2个次要成分进行了鉴定 .结果证明 ,主要成分即为目标九肽 ;先于目标肽洗脱的副产物分子量比目标肽的分子量多 16 .MS MS证明 ,该副产物包括 2种九肽衍生物 :1种为其Tyr1氧化生成 ,另 1种为Met6氧化生成 ;后于目标肽洗脱的副产物为九肽的Tyr1残基增加 12所致 ,这种现象尚未见报道 .对副产物形成的可能机制进行了讨论
A solid-phase chemically synthesized nonapeptide H2N Tyr Val Asn Val Asn Met Gly Leu Lys CONH2 and its two major byproducts were analyzed by tandem mass spectrometry (RPHPLC MS MS) coupled to reversed-phase high performance liquid chromatography.According to Fmoc Chemistry, stepwise coupling of nonapeptide synthesis on Fmoc PAL PEG · PS resin from C-terminus to N-terminus.After conjugation was complete, reagents R (90% TFA, 5% thioanisole, 3% dimercaptoethanol Alkane, 2% anisole) at room temperature (20 ~ 25 ℃) for 2 5h, the peptide was deprotected and the resin was removed.PRP HPLC analysis showed that the crude product contained one major component, two minor components And several trace components.The main components and two minor components were identified by RPHPLC MS MS respectively.The results showed that the main component was the target nonapeptide.The molecular weight of the byproduct before the target peptide eluted was higher than that of the target peptide More 16. MS MS demonstrated that the by-products include two nonapeptide derivatives: one for its Tyr1 oxidation and the other for Met6 oxidation; the by-product for subsequent target peptide elution is the non-peptide Tyr1 residue An increase of 12 due to this phenomenon has not been reported on the possibility of the formation of by-products were s