Table 2.

¹H-NMR spectral data of glycerol (1) in D₂O solutions (5~540 mM, 19–21 °C) without DSS and the proportions of the three rotamers (%) determined in the current ¹H-NMR Karplus analysis using the two types of equations, Equation (1) (eq 1) and Equation (2) (eq 2).

Entry	Solvent (mM)	1H NMR Spectral Data d ppm/J (Hz)			Chemical Shift Difference (D ppm) (Entry X − Entry 1)			Rotamers (%) 2
								by Equation (1)
								/Equation (2)
		H2	H1S/H3R	H1R/H3S	DH2	DH1S	DH1R	gt (α)	gg (γ)	tg (β)
1	D2O (5 mM)	3.727	3.595	3.505	0	0	0	51	32	17
		m	4.3	6.6, 11.7				(50	27	23)
2	D2O (54 mM)	3.721	3.589	3.499	−0.006	−0.006	−0.006	51 (50	32 27	17 23)
		m	4.3	6.6, 11.7
3	D2O (108 mM)	3.719	3.587	3.497	−0.008	−0.008	−0.008
		m	4.3	6.6, 11.7
4	D2O (540 mM)	3.711	3.58	3.489	−0.016	−0.015	−0.016
		m	4.3	6.6, 11.7
5	D2O (+H2O) (500 mM) 1	3.719	3.587	3.497	−0.008	−0.008	−0.008	51	32	17
		m	4.3	6.6, 11.7				(50	27	23)

¹ After treatment with 10 molar excess amounts of H₂O for 2 h, the aqueous glycerol was diluted with D₂O to 500 mM and applied to the ¹H-NMR spectroscopic measurement as shown in Figure 4 (500 MHz at 20.7 °C). ² Populations (%) are calculated with Equation (1) (60, −60, 180 in degrees) and Equation (2) (65, −65, 180 in degrees) that are given in System of Equations in the preceding section of Materials and Methods.