Table 2.
LC-MS/MS conditions and parameters for program 1
| Name | MRM | Dwell (s) | CV (V) | CE (V) | Mode | Retention time (min) | Time window ± (min) | Performance |
|---|---|---|---|---|---|---|---|---|
| Norvaline | 117.9 → 54.9 | 0.005 | 24 | 16 | ESI+ | 7.2 | 0.2 | ∗∗ |
| Tryptophan (Trp) | 205.1 → 118.0 | 0.005 | 22 | 26 | ESI+ | 7.4 | 0.2 | ∗∗ |
| Tyrosine (Tyr) | 182.0 → 136.1 | 0.005 | 20 | 12 | ESI+ | 8.0 | 0.15 | ∗∗ |
| Arginine (Arg) | 175.1 → 70.0 | 0.005 | 34 | 20 | ESI+ | 10.6 | 0.4 | ∗∗∗ |
| Phenylalanine (Phe) | 166.0 → 120.1 | 0.005 | 24 | 12 | ESI+ | 7.1 | 0.3 | ∗∗ |
| Histidine (His) | 156.0 → 110.0 | 0.005 | 26 | 14 | ESI+ | 10.7 | 0.5 | ∗∗∗ |
| Methionine (Met) | 150.0 → 56.0 | 0.005 | 22 | 16 | ESI+ | 7.6 | 0.2 | ∗∗ |
| Glutamic acid (Glu) | 148.0 → 84.0 | 0.005 | 20 | 16 | ESI+ | 8.8 | 0.35 | ∗∗∗ |
| Lysine (Lys) | 147.0 → 67.0 | 0.005 | 22 | 26 | ESI+ | 10.8 | 0.3 | ∗∗∗ |
| Glutamine (Gln) | 147.0 → 41.0 | 0.005 | 22 | 24 | ESI+ | 9.3 | 0.35 | ∗∗ |
| Aspartic acid (Asp) | 134.0 → 73.9 | 0.005 | 20 | 12 | ESI+ | 9.1 | 0.4 | ∗ |
| Asparagine (Asn) | 133.0 → 73.9 | 0.005 | 20 | 14 | ESI+ | 9.4 | 0.35 | ∗∗∗ |
| Isoleucine (Ile) | 132.0 → 69.0 | 0.005 | 20 | 16 | ESI+ | 7.3 | 0.25 | ∗∗ |
| Leucine (Leu) | 132.0 → 43.4 | 0.005 | 20 | 22 | ESI+ | 7.0 | 0.2 | ∗∗ |
| Cysteine (Cys) | 122.0 → 75.9 | 0.005 | 18 | 12 | ESI+ | 8.2 | 0.35 | ∗ |
| Threonine (Thr) | 120.0 → 56.0 | 0.005 | 48 | 14 | ESI+ | 8.7 | 0.35 | ∗∗∗ |
| Valine (Val) | 118.0 → 72.0 | 0.005 | 20 | 10 | ESI+ | 7.7 | 0.3 | ∗∗ |
| Proline (Pro) | 116.0 → 70.0 | 0.005 | 30 | 12 | ESI+ | 7.8 | 0.4 | ∗∗ |
| Serine (Ser) | 106.0 → 60.0 | 0.005 | 22 | 8 | ESI+ | 9.2 | 0.4 | ∗∗∗ |
| Alanine (Ala) | 90.0 → 44.0 | 0.005 | 20 | 8 | ESI+ | 8.4 | 0.2 | ∗∗ |
| Glycine (Gly) | 75.9 → 29.8 | 0.005 | 18 | 6 | ESI+ | 8.8 | 0.3 | ∗∗∗ |
| Glutathione (GSH) | 308 → 179 | 0.005 | 20 | 10 | ESI+ | 8.4 | 0.2 | ∗∗∗ |
| Glutathione disulfide (GSSG) | 613 → 355 | 0.005 | 40 | 20 | ESI+ | 11.5 | 0.1 | ∗∗∗ |
| Ascorbic acid | 175 → 115 | 0.005 | 40 | 10 | ESI- | 4.4 | 0.25 | ∗ |
| S-Adenosyl-L-methionine (SAM) | 399 → 250 | 0.005 | 20 | 10 | ESI+ | 11.3 | 0.2 | ∗∗ |
| S-Adenosyl-L-homocysteine (SAH) | 385 → 136 | 0.005 | 20 | 20 | ESI+ | 9.8 | 0.1 | ∗∗ |
| Myo-inositol | 179.2 → 86.9 | 0.005 | 40 | 12 | ESI- | 9.1 | 0.1 | ∗∗ |
Note: Italicized metabolites are referenced from a database (https://metlin.scripps.edu) but not confirmed with standards. CV: cone voltage; CE: collision energy; Performance is evaluated based on how easily peak identification could be done with 10 μL injected hESC sample that was extracted as described in Method 1. ∗∗∗, single peak, easily identified; ∗∗, multiple peaks, target peak could be identified based on the retention time of the standard or by comparing the isotopic ratio, and target peak is not overlapped with other peaks; ∗, multiple peaks, very weak (below 3 folds of baseline) target peak intensity and/or target peak overlaps with other peaks.