高效液相色谱-电喷雾-质谱法测定蛋白质混合物

Determination of Protein Mixture by High PerformanceLiquid Chromatography Electrospray IonizationMass-spectrometry (HPLC-ESI-MS)

介绍高效液相色谱 -电喷雾 -质谱 (HPL C- ESI- MS)联用技术在生物大分子 (蛋白 )的分离和检测上所表现出来的优越性能 ,并利用 L C- ESI- MS技术成功地分离、检测溶菌酶和牛血清蛋白混合物 .

A rapid and precise method for the determination of lysozyme and bovine serum al bumin in protein mixture for specific purposes, based on high performance liquid chromatography electrospray ionization mass spectrometry, has been developed. The compound was identified using mass spectrometry. In HPLC experim ent the column was a Spherisorb C 18 reversed phase column, 10 μm, 300  , 4.6 mm×250 mm; the mobile phase A (10% acetonitrile, 0.1% trifluoroacetic aci d); the mobile phase B (90% acetonitrile, 0.1% trifluoroacetic acid); gradient e lution, mobile phase B changed from 20% to 100% in 40 minutes; flow rate, 0.2 m L/min.The experimental re sults showed that the proteins lysozyme and bovine serum albumin were well separ ated in HPLC, and were well determined by mass detector. The quantitation was pe rformed with external standard.