The band-selective13C pulse calibrations should be done in the inverse mode (13C through the 2nd channel) using the pulse sequence decp90sp and decp180sp, respectively. (see Tutorial: Pulse Calibrations). You also calculate the values by using the shape tool in XWINNMR, stdisp. For instance, a sample of 0.1 M CH3OH-13C in DMSO-d6. can be used. The 13C-offset for the methyl group in methanol is 49 ppm, the methyl 1H-offset is 3.28 ppm and the one-bond coupling constant is 142 Hz. The calibration affords an antiphase signal that its intensity depends on the flip angle of the selective 13C-pulse. When the 13C-pulse is a perfect 90-pulse, the magnetization is completely converted to a multiple quantum coherence and the obseved signal reaches its minimum.You can also calculate the values by using the shape tool in XWINNMR, stdisp
Selective excitation
Acquisition parameters for band-selective 13C 90 degree pulse calibration.
PULPROG: decp90sp NUC1: 1H nucleus on f1 channel O1P: 3.5 ppm 1 H offset NUC2: 13C nucleus on f2 channel O2P: 49 ppm 13C: offset NS: 1 DS: 0 CNST2: 139 PL2: 120dB P13: selective pulse length (409us, e.g.) SP2: dB power level for selective pulse SPNAM2: G4.256 SPOAL2: 0.5 SPOFF2: 0.0
13C 90deg shaped pulse name (spnam2=G4.256), length (p13=409u), and power (sp2) applied on-resonance. When applied off-resonance, the pulse is specified with sp4, p13=409u and spnam4=G4.256. 13C 90deg time-reversed shaped pulse name (spnam8=G4tr.256), length (p13), and power (sp8)
Selective inversion
Acquisition parameters for band-selective 13C 1800 degree pulse calibration. If the selective 13C-pulse, applied after the 180- 1H-pulse, is a perfect 180-pulse, the magnetization stays as multiple quantum coherence and the observable signal is minimal.
PULPROG: decp180sp pulse program NUC1: 1H nucleus on f1 channel O1P: 3.5 ppm 1 H offset NUC2: 13C nucleus on f2 channel O2P: 49 ppm 13C: offset NS: 2 DS: 2 CNST2: 139 PL2: 120dB P13: selective pulse length (256us, e.g.) SP3: dB power level for selective pulse SPNAM3: G3.256 SPOAL3: 0.5 SPOFF3: 0.0
13C 180deg shaped pulse name (spnam3=G3.256), length (p14=256u), and power (sp3) usually applied on CA/CB resonances. Otherwhise, when applied off-resonance, is specified with sp5/sp7. 13C 180deg shaped pulse name applied exclusively on CA resonances (spnam9), length p24=768u, and power (sp9) Selective decoupling
Selective decoupling is applied on the alpha-carbon region (centered at 52 ppm) during the constant time evolution of the 15N-magnetization, in order to ensure that the scalar coupling between 13C a and amide 15N does not evolve. The frequency offset is given through the parameter SPOFF15 which should be given a negative value, in Hz, corresponding to the difference 173-52 ppm=121 ppm of 13C at the field you are working at.
13C 180deg decoupling shaped pulse name applied exclusively on CA resonances (spnam15=g3.256), length pcpd2=768u, and power (sp15=pl12) using cpdprg2=mlevsp180 This is an example of carbon pulses defined in edprosol: