摘要
目的探讨纳升超高效液相色谱-飞行时间质谱（nano—UPLC—TOF—MS）检测斑马鱼胚胎中小分子多肽的方法。方法收集受精后发育24h的斑马鱼胚胎，去除外层胶膜，采用机械力破碎法去除斑马鱼早期胚胎中的卵黄，并用显微镜和聚丙烯酰胺凝胶电泳检测去除效果，然后用小分子肽提取液获得肽液样品后，用nano—UPLC—TOF—MS检测小分子肽相对分子质量。结果经处理的胚胎卵黄几乎完全被去除，nano．UPLC—TOF—MS结果显示在22～24．5min处有一明显的样品峰，单质量／电荷比（m／z）显示，绝大部分肽段峰位于800～1400m／z范围内。结论Nano—OPLC—MS是检测小分子多肽的有效方法，早期斑马鱼胚胎中存在着小分子多肽，其相对分子质量集中在800～1400范围内。
Objective To apply the nano-ultra high performance liquid chromatography coupling with time- of-fllght mass spectrometer （nano-UPLC-TOF-MS） to detect the low molecular weight polypeptides in zebraflsh embryos. Methods Zebrafish embryos were collected 24 hours after fertilization and the outer chorlon was removed. And then the yolk was removed with mechanical forces, of which the efficiency was validated by microscope and SI）S-PAGE; the low molecular weight polypeptide was extracted with corresponding solution, and followed by separation and detection with nano-UPLC-TOF-MS. Results Mechanical vortex almost completely removed yolk cells of zebrafish embryos, from which the special solution extracted effectively low molecular weight polypeptides for the later detection. As demonstrated by nano-UPLC-TOF-MS, the polypeptide peak showed between 22 and 24.5 min, and the m/z ration of most of polypeptides were between 800-1400. Conclusion Nano- OPLC-TOF-MS is effective to detect low molecular weight polypeptides, and low molecular weight peptides definitely exist in zebraflsh embryos, of which the relative molecular mass seems to be 800 to 1400, probably consists of about 10 residues.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2007年第6期1033-1036,共4页
Journal of Sichuan University(Medical Sciences)
