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目的对一种小分子生物活性肽(重组BMP)样品进行电喷雾质谱测定,初步确定其一级结构。方法离子化方式:电喷雾离子化;检测方式正离子;质量扫描范围m/z 150～1 000;干燥气(N2)流速5 arb;雾化气(N2)压力35 arb;毛细管出口电压4.5 kV;碎裂器电压100 V;碰撞诱导解离技术碎裂器电压为35%;碰撞气流量为电子流量控制器自动控制;进样方式:流动注射进样;流动相为乙腈∶水(50∶50)。结果利用电喷雾质谱技术,初步测定重组BMP样品的一级结构为H-Lys-Gly-Asp-Glu-Glu-Ser-Leu-Pro-OH。结论该方法能够初步测定小分子生物活性肽的一级结构,为实验室生物工程菌构建提供准确有效的检测技术。
Objective To determine the primary structure of a sample of small molecule bioactive peptides (recombinant BMP) by electrospray ionization mass spectrometry. Methods ionization: electrospray ionization; detection mode positive ions; mass scan range m / z 150-1000; dry gas (N2) flow rate 5 arb; atomization gas (N2) pressure 35 arb; capillary exit voltage 4.5 kV ; Fragmentation voltage 100 V; collision-induced dissociation fragmentation voltage 35%; collision gas flow control for the electronic flow controller; injection mode: flow injection injection; mobile phase acetonitrile: water 50). Results The primary structure of recombinant BMP was determined by electrospray ionization mass spectrometry (H-Lys-Gly-Asp-Glu-Glu-Ser-Leu-Pro-OH). Conclusion The proposed method can be used to determine the primary structure of small molecular bioactive peptides and provide an accurate and effective detection technique for the construction of laboratory bioengineered bacteria.