1：Experimental Design and Method Selection:

The study used IRMPD spectroscopy, drift-tube ion-mobility mass spectrometry, and DFT calculations to investigate chiral differentiation. IRMPD was performed in the 2650-3800 cm?1 range, and ion-mobility measured collision cross-sections. DFT calculations at wB97X-D/6-31G* and 6-311G** levels were used for structural and spectral analysis.

2：Sample Selection and Data Sources:

Samples included L-Ile, D-Ile, permethylated β-cyclodextrin, acetic acid, water, and methanol. Solutions were prepared with 20 mM concentrations in a mixture of H2O:CH3OH:acetic acid (49:49:2 v/v/v).

3：List of Experimental Equipment and Materials:

FT-ICR mass spectrometer (Ionspec), OPO lasers (IROPO from OPOTek and home-built), Nd:YAG laser (Quantel), Agilent 6560 ion mobility quadrupole time-of-flight mass spectrometer, Gaussian 09 software, CP2K code, MOBCAL program, Sigma code. Materials from Sigma-Aldrich and Sejin Chemical Industry Co.

4：Experimental Procedures and Operational Workflow:

Non-covalent complexes were formed via electrospray ionization, isolated using SWIFT, and irradiated with IR lasers. IRMPD spectra were acquired by monitoring ion abundance reduction. Ion-mobility experiments used helium buffer gas at specific pressures and voltages to measure arrival times and CCS.

5：Data Analysis Methods:

IR spectra were power-normalized and smoothed. CCS values were calculated using PA, EHSS, TM, and modified TM methods. DFT calculations included optimization, frequency analysis, and Gibbs free energy estimation.