质谱的讲义44.pptVIP

第四节、 串联质谱应用 一、碰撞诱导裂解（CAD） 二、源内CAD 三、IRMPD和ECD 一、碰撞诱导裂解（CAD） 二、源内CAD 三、IRMPD和ECD Infrared Multiphoton Dissociation: IRMPD Electron Capture Dissociation: ECD ECD 9 eV [M+nH]n+ + e- [M+nH](n+1)+· + 2e- [M-nH]n- + e- [M-nH](n-1)-· + 2e- [M-nH]n- + e- ([M-nH](n-1)-·)*transient + 2e- fragments + e- More energetic electrons may cause double ionization and Coulombic explosion. * API 4000 LC/MS/MS APCI ESI Linear Varian 1200 GC/MS/MS 180o Magnet ESI source capillary ion guide cell Differential pumping stages CO2 - Laser HPLC Setup of IRMPD fast MS/MS coupled to HPLC Infrared Multiphoton Dissociation (IRMPD) Hot topic at this year‘s ASMS !! N N O N H S O H S O O N H N S O O r N H 2 O N O s ?m = 1.1 ppm ?m = 0 ppm ?m = 0 ppm ?m = 0.9 ppm ?m = 1.4 ppm ?m = 0.6 ppm 2 3 6s 1 4 5 6 1 3 4 6 5 SORI-CAD MS/MS TIC IRMPD MS/MS TIC 2 Infrared Multiphoton Dissociation (IRMPD) Applications for IRMPD MS/MS with HPLC No need for pulsed gas, faster MS/MS experiment High throughput proteomics Rapid acquisition of MS/MS fragment information from peptides with accurate mass Singly charged peptides from MALDI are often difficult to fragment by CID in FTMS General Structural Characterization of Natural Products, Drug Candidates, Oligonucleotides, Other Unknowns IRMPD gives more extensive fragmentation with many compound classes 150 200 250 300 350 400 450 500 550 600 650 m/z SORI-CAD IRMPD Conversion efficiency: 98% time to acquire single scan: 3s Conversion efficiency: 67% time to acquire single scan: 1s MH+ MH+ Sample: reserpine Mass spec.: 3.0 T FTMS Compare IRMPD with “normal” SORI CID - Ions Present 2.0 2.5 3.0 3.5 min 1 2 3 4 5 6 2.0 2.5 3.0 3.5 min 1 2 3 4 5 6 IRMPD TIC Acquisition rate = 1 Hz CAD TIC Acquisition rate = 0.3 Hz Compare IRMPD with “normal” SORI CID - Duty Cycle 200 300 400 500 600 700 800 900 1000 1100 1200 m/z MALDI FTMS of LHRH IRMPD MS/MS of the MH+ IR laser: 200ms @ 50% power b2 y6 y5