Acquisition and processing of the 1H NMR spectra

The experiments were performed without spinning at 298 K using a Bruker Avance III of 9.4 T (400.1819 MHz, ¹H frequency) spectrometer equipped with a 5 mm TBI probe. Both T₂-edited and diffusion-edited ¹H NMR spectra were acquired for each sample. The T₂-edited ¹H NMR spectra were acquired with Carr–Purcell–Meiboom–Gill pulse sequence (CPMG) to suppress plasma protein, and lipoprotein signals with T₂. PRESAT was used to suppress the water signal. The following parameters were applied: 300 μsec of evolution time and 600 msec of total spin–spin relaxation delay and 64 scans. The magnetic field homogeneity was optimized for each sample. The diffusion-edited ¹H NMR spectra were acquired by bipolar pulse pair-longitudinal eddy current delay pulse sequence that was intended to suppress signals from molecules with large diffusion coefficients, which are the small molecules. The WATERGATE 3-9-19 pulse sequence was used to suppress the water signal. A sine-shaped gradient pulse of 90% maximum strength, with 2.0 msec of duration (little delta), followed by a delay (big delta) of 100 msec that allowed the eddy currents to decay, was applied in the spectra acquisition. The delay for the binomial water suppression was set at 17.0 μsec, the gradient pulse was 1.0 msec, and the spectra were acquired with 32 scans. For both pulse sequences, the remaining acquisition parameters were as follows: spectral width, 6.0 kHz; data size, 32 k; acquisition time, 2.73 sec; constant receiver gain, 203; and relaxation delay, 5.0 sec. The data processing included zero filling to 128 k, line-broadening multiplication by 1.0 (T₂-edited ¹H NMR) and 3.0 Hz (diffusion-edited ¹H NMR), and Fourier transformation. All of the spectra were phase and baseline corrected and referenced to TMSP at 0.000 ppm (T₂-edited ¹H NMR) and to the methyl resonance of lipoproteins at 0.800 ppm (diffusion-edited ¹H NMR) using TopSpin software (v3.1; Bruker BioSpin). Typical full spectra are presented as Supplementary Data (Supplementary Figs. S1 and S2).

NMR peak assignment was performed by means of literature data (Bell et al., 1987; Fan, 1996; Foxall et al., 1993) and confirmed with heteronuclear single quantum coherence spectroscopy (2D ¹H-¹³C HSQC) and selective homonuclear total correlation spectroscopy (selective 1D ¹H-¹H TOCSY) using Bruker's available sequences.