CH Mode Mixing Determines the Band Shape of the Carboxylate Symmetric Stretch in Apo-EDTA, Ca2+–EDTA, and Mg2+–EDTA

The infrared spectra of EDTA complexed with Ca²⁺ and Mg²⁺ contain, to date, unidentified vibrational bands. This study assigns the peaks in the linear and two-dimensional infrared spectra of EDTA, with and without either Ca²⁺ or Mg²⁺ ions. Two-dimensional infrared spectroscopy and DFT calculations reveal that, in both the presence and absence of ions, the carboxylate symmetric stretch and the terminal CH bending vibrations mix. We introduce a method to calculate participation coefficients that quantify the contribution of the carboxylate symmetric stretch, CH wag, CH twist, and CH scissor in the 1400-1550 cm^-1 region. With the help of participation coefficients, we assign the 1400-1430 cm^-1 region to the carboxylate symmetric stretch, which can mix with CH modes. We assign the 1000-1380 cm^-1 region to CH twist modes, the 1380-1430 cm^-1 region to wag modes, and the 1420-1650 cm^-1 region to scissor modes. The difference in binding geometry between the carboxylate-Ca²⁺ and carboxylate-Mg²⁺ complex manifests as new diagonal and cross-peaks between the mixed modes in the two complexes. The small Mg²⁺ ion binds EDTA tighter than the Ca²⁺ ion, which causes a redshift of the COO symmetric stretches of the sagittal carboxylates. Energy decomposition analysis further characterizes the importance of electrostatics and deformation energy in the bound complexes.