## 3 Results  
Altogether, 24 metabolites were identified in denatured plasma in both patients and healthy subjects, where the signals from 19 compounds were sufficient for quantitative evaluation (Table 2). Further in the text, we use the trivial names of 2-ketoads derived from leucine, isoleucine, and valine (IUPAC names are in Table 2). Besides molecular metabolites, we also evaluated the lipoprotein fraction, which, as described by [PERSON] et al., contains very-low-density lipoproteins (VLDL), low-density lipoproteins (LDL), and high-density lipoproteins (HDL), including up to one-third of triacylglycerideys [18]. For multivariate analyses, we used the relative concentrations of plasma metabolites (expressed as the integral of a particular spectral region) as an input in order to target biologically informative value. We avoided feeding the algorithms with binned NMR spectra as is common in metabolomic studies, since there may be regions of NMR spectra marked as important that are not straightforward and unambiguously related to biological relevance. Firstly, the data of all patients were analysed (Group A and Group B together) on Day 1 against controls by PCA and PLS-DA (Figure 1). In contrast to patients, controls were relatively clustered together. The loading values were the highest for glucose, 3-hydroxybutyrate, and leucine in PC1 and alanine, lactate, and glutamine in PC2. The situation was very similar after the PLS-DA run. The 10-fold cross-validated PLS-DA algorithm performed with accuracy of 0.954, R2 of 0.7926, and Q2 of 0.6749 for eight components. The variables with the highest VIP scores were: glucose, 3-hydroxybutyrate, alanine, leucine, valine, and glutamine (performance measured in accuracy). The incorporation of additional variables did not improve the performance. \begin{table}
\begin{tabular}{c c} \hline
**Metabolite** & **NMR Peak Assignment, Confirmed by [PERSON] and [PERSON]** \\ \hline lactate & 1.33 (d; J = 7.0), 4.12 (q; J = 7.0) \\ glutamine & 2.12 (m), 2.15 (m), 2.44 (m), 2.48 (m), 3.77 (dd) \\ isoleucine & 0.94 (t; J = 7.5), 1.01 (d; J = 7.0), 3.68 (d; J = 4.2) \\ leucine & 0.96 (d; J = 6.2), 0.97 (d; J = 6.1), 1.68 (m), 1.72 (m), \\ phenylalanine & 3.13 (m), 3.28 (m), 7.34 (d; J = 7.5), 7.38 (t; J = 7.4), 7.44 (t) \\ tyrosine & 3.05 (dd), 3.20 (dd), 3.93 (dd), 6.91 (d; J = 8.5), \\  & 7.20 (d; J = 8.5) \\ valine & 0.99 (d; J = 7.1), 1.04 (d; J = 7.1), 2.27 (m), 3.61 (d; J = 4.4) \\ pyruvate & 3.28 (s) \\ citrate & 2.54 (d), 2.67 (d) \\ acetate & 1.92 (s) \\ alanine & 1.48 (d; J = 7.30), 3.78 (q) \\ glucose & 3.23 (m), 3.40 (m), 3.46 (m), 3.52 (dd), 3.78 (m), 3.82 (m), \\  & 3.89 (dd), 4.64 (d), 5.23 (d) \\
3-hydroxybutyrate & 1.20 (d; J = 6.23 Hz), 2.31 (m), 2.41 (m), 4.16 (m) \\ creatine & 3.04 (s), 3.94 (s) \\ lysine & 1.33 (d), 3.58 (d; J = 4.9), 4.25 (m) \\
2-oxoisocoronate (2-ketoleucine) & 0.94 (d; J = 6.6), 2.11 (m), 2.61 (d; J = 7.0) \\ \(\alpha\)-ketooisomerase (2-ketovaline) & 1.11 (d; J = 7.1), 3.01(dq) \\
3-methyl-2-oxo-valerate & 0.90 (t; J = 7.5), 1.10 (d; J = 6.7) \\ (2-ketoisoleucine) & \\ lipoprotein fraction & 0.82–0.93 (m), 1.20–1.37 (m) \\ \# creatinine & 3.05 (s), 4.07 (s) \\ \# histidine & 7.07 (s), 7.80 (s) \\  & 1.46 (m), 1.50 (m), 1.73 (m), 1.89 (m), 1.93 (m), \\  & 3.03 (t; J = 7.6) \\ \# threonine & 1.34 (d), 3.56 (d; J = 4.9),4.26 (m) \\ \# tryptophan & 7.21 (t), 7.30 (t), 7.33 (s), 7.56 (d), 7.74 (d; J = 8.0) \\ \hline \end{tabular}
\end{table} 
Table 2: Chemical shifts (in ppm), J couplings (in Hz), and multiplicities (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; dd, doublet of doublets; dq, doublet of quartets) for the pool of metabolites identified in blood plasma. Signals marked with # were not suitable for quantitative analyses.