Table 1.
Summary of method validation parameters for 26 NMF.
| NMF | MRM or SIM condition |
SIL-IS | MRM or SIM condition |
Dynamic range [nmol/cm2] |
Slope ± SD | Intercept ± SD | R2 | %RSD | LOQ [nmol/cm2] |
||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Transition | EV | Transition | EV | NMF conc. [nmol/cm2] |
25 | 5 | 2.5 | 1.25 | 0.5 | 0.25 | 0.1 | 0.05 | |||||||
| Val | 118.10 > 72.20 (+) | -12 | ValSIL | 119.10 > 73.15 (+) | −11 | 0.05–25 | 7.9 ± 0.34 | 3.9 ± 0.59 | 0.998 | 4 | 5 | 9 | 7 | 13 | 10 | 2 | 14 | 0.23 | |
| Asn | 131.05 > 113.10 (−) | 14 | AsnSIL | 133.10 > 115.10 (−) | 16 | 0.05–5 | 8.0 ± 0.21 | −0.39 ± 0.039 | 0.979 | — | 16 | 14 | 4 | 2 | 11 | 6 | 10 | 0.60 | |
| Asp | 132.05 > 115.05 (−) | 15 | AspSIL | 137.00 > 92.05 (−) | 13 | 0.05–5 | 1.4 ± 0.1 | 0.43 ± 0.074 | 0.964 | — | 7 | 12 | 3 | 6 | 11 | 10 | 5 | 0.57 | |
| Cit | 174.10 > 131.10 (−) | 13 | AsnSIL | 133.10 > 115.10 (−) | 16 | 0.05–2.5 | 0.69 ± 0.042 | −0.013 ± 0.023 | 0.991 | — | — | 15 | 14 | 11 | 3 | 7 | 5 | 0.86 | |
| Cys | 122.05 (+) | — | MetSIL | 151.10 > 61.05 (+) | −22 | 0.5–5 | 1.8 ± 0.26 | 0.93 ± 0.15 | 0.958 | — | 15 | 31 | 9 | 8 | — | — | — | 2.6 | |
| Gln | 147.10 > 84.10 (+) | −18 | AsnSIL | 135.10 > 75.10 (+) | −15 | 0.05–5 | 43 ± 1.5 | 0.2 ± 2.5 | 0.987 | — | 5 | 11 | 14 | 5 | 13 | 10 | 20 | 0.38 | |
| Glu | 146.05 > 102.10 (−) | 15 | GluSIL | 147.10 > 103.10 (+) | −15 | 0.25–5 | 1.8 ± 0.26 | 2.0 ± 0.31 | 0.833 | — | 13 | 15 | 13 | 11 | 15 | — | — | 1.2 | |
| Gly | 76.1 (+) | — | AlaSIL | 91.10 > 45.10 (+) | −15 | 0.5–5 | 3.5 ± 0.67 | −1.3 ± 0.31 | 0.974 | — | 19 | 12 | 4 | 15 | — | — | — | 3.7 | |
| His | 156.10 > 110.05 (+) | −15 | HisSIL | 159.10 > 113.10 (+) | −15 | 0.05–25 | 3.4 ± 0.29 | 3.3 ± 0.74 | 0.99 | 8 | 11 | 17 | 14 | 18 | 8 | 19 | 6 | 0.34 | |
| Ile | 132.10 > 69.10 (+) | −16 | LeuSIL | 133.10 > 87.10 (+) | −11 | 0.05–25 | 1.0 ± 0.19 | −0.15 ± 0.28 | 0.998 | 18 | 4 | 8 | 6 | 17 | 16 | 10 | 8 | 0.46 | |
| Leu | 132.10 > 43.10 (+) | −26 | LeuSIL | 133.10 > 87.10 (+) | −12 | 0.05–25 | 0.5 ± 0.063 | 0.24 ± 0.11 | 0.999 | 12 | 14 | 25 | 11 | 10 | 10 | 3 | 22 | 0.20 | |
| Lys | 147.10 > 67.15 (+) | −25 | LysSIL | 149.10 > 57.15 (+) | −25 | 0.05–5 | 5.6 ± 0.69 | 0.33 ± 0.53 | 0.938 | — | 13 | 34 | 5 | 6 | 13 | 11 | 11 | 0.43 | |
| Met | 150.05 > 56.15 (+) | −17 | MetSIL | 151.10 > 61.05 (+) | −22 | 0.05–5 | 8.7 ± 0.33 | −0.016 ± 0.66 | 0.993 | — | 6 | 14 | 12 | 4 | 9 | 24 | 12 | 0.37 | |
| Orn | 133.10 > 70.15 (+) | −17 | LysSIL | 149.10 > 67.15 (+) | −25 | 0.05–2.5 | 120 ± 11 | 9.7 ± 5.3 | 0.928 | — | — | 7 | 3 | 9 | 8 | 8 | 10 | 0.70 | |
| PCA | 128.10 > 84.05 (−) | 14 | PCASIL | 133.10 > 89.10 (−) | 15 | 0.05–5 | 1.7 ± 0.12 | 0.64 ± 0.11 | 0.978 | — | 7 | 10 | 8 | 4 | 18 | 6 | 3 | 0.12 | |
| Phe | 166.10 > 120.05 (+) | −15 | PheSIL | 171.10 > 125.15 (+) | −15 | 0.05–5 | 8.9 ± 0.1 | 1.4 ± 0.2 | 0.992 | — | 2 | 4 | 7 | 9 | 7 | 8 | 13 | 0.073 | |
| Pro | 116.05 > 70.10 (+) | −16 | ProSIL | 117.10 > 71.10 (+) | −16 | 0.05–25 | 5.1 ± 0.41 | −0.16 ± 0.83 | 0.999 | 8 | 22 | 42 | 13 | 7 | 6 | 14 | 17 | 0.22 | |
| Ser | 106.05 > 60.05 (+) | −11 | SerSIL | 105.10 > 74.10 (−) | 15 | 0.05–25 | 31 ± 1.7 | −0.033 ± 0.39 | 0.899 | 21 | 12 | 18 | 5 | 15 | 3 | 19 | 4 | 0.15 | |
| Thr | 120.05 > 56.00 (+) | −15 | AsnSIL | 133.10 > 115.10 (−) | 16 | 0.05–5 | 4.0 ± 0.21 | 2.2 ± 0.23 | 0.896 | — | 5 | 8 | 11 | 6 | 6 | 5 | 11 | 0.14 | |
| Trp | 205.10 > 188.10 (+) | −11 | TrpSIL | 207.10 > 189.00 (+) | −10 | 0.05–5 | 2.9 ± 0.51 | 0.82 ± 0.51 | 0.835 | — | 16 | 56 | 12 | 18 | 14 | 12 | 8 | 0.37 | |
| Tyr | 182.10 > 91.10 (+) | −28 | TyrSIL | 183.10 > 137.15 (+) | −13 | 0.05–5 | 8.8 ± 1.3 | 0.43 ± 1 | 0.993 | — | 16 | 35 | 4 | 8 | 18 | 8 | 14 | 0.37 | |
| UCA | 137.10 > 93.05 (−) | 14 | AspSIL | 137.00 > 92.05 (−) | 13 | 0.05–5 | 4.8 ± 0.43 | 2.1 ± 0.17 | 0.921 | — | 8 | 10 | 8 | 8 | 5 | 9 | 6 | 0.32 | |
| LA | 89.1 (−) | — | LASIL | 92.1 (−) | — | 0.5–5 | 1.6 ± 0.26 | 0.96 ± 0.18 | 0.974 | — | 15 | 24 | 5 | 5 | — | — | — | 3.9 | |
| Urea | 61.0 (+) | — | UreaSIL | 62.0 (+) | — | 0.25–5 | 0.39 ± 0.038 | 0.13 ± 0.024 | 0.948 | — | 10 | 14 | 11 | 10 | 6 | — | — | 1.2 | |
| Arg | 175.10 > 70.10 (+) | −17 | LysSIL | 149.10 > 57.15 (+) | −25 | 0.25–5 | 0.51 ± 0.035 | −0.084 ± 0.18 | 0.97 | — | 7 | 11 | 15 | 12 | 18 | — | — | 1.7 | |
| Ala | 88.0 (−) | — | AlaSIL | 91.10 > 45.10 (+) | −15 | 0.05–5 | 2.3 ± 0.21 | 0.38 ± 0.8 | 0.969 | — | 9 | 16 | 9 | 6 | 8 | 10 | 15 | 0.34 | |
The actual concentration values used are 0.05, 0.1, 0.25, 0.5, 1.25, 2.5, 5, 25 nmol/cm2. Amino acids are described with three-letter abbreviations. Cit, citrulline; Orn, ornithine; PCA, pyrrolidone carboxylic acid; UCA, urocanoic acid; LA, lactic acid; (+), positive ion mode; (−), negative ion mode; EV, electrical voltage; R2, correlation coefficient; LOQ, limit of quantification.