质谱讲义4-4（解析-串联）.PPT.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） API 4000 LC/MS/MS APCI Linear Varian 1200 GC/MS/MS 180o 二、源内CAD 三、IRMPD和ECD Infrared Multiphoton Dissociation: IRMPD Electron Capture Dissociation: ECD 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 !! 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 Compare IRMPD with “normal” SORI CID - Ions Present 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 y4 y3 b6 b7 b8 b4 b3 b5–H2O b4–H2O MH+ MH+-H2O IRMPD Data Examples (2) 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. Remaining +2 Parent Ion After ECD c10 c6 c7 c8 c9 c5 C Sequence Ions in the ECD Spectrum of Substance P c25 C21 c22 c23 c24 C15 C18 C12 C11 C10 C7 C6 z24 z23 z20 z19 z17 z16 z14 z12 z9 z5 z3 C and Z Sequence Ions in the ECD Spectrum of Melittin