Table 1.
Method validation characteristics in serum
| Compound | Linearitya (R2) | LOD (nM) | Linear range (nM) | RT stability (n = 3)b (RSD, %) | Precision in serum (RSD, %) | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| Intraday | Interday | |||||||||
| C2–low | C4–medium | C7–high | C2–low | C4–medium | C7–high | |||||
| Lysophosphatidic acid classes | ||||||||||
| aLPA C18:1 | 0.9963 | 1 | 4-533 | 0.79 | 19.9 | 8.8 | 27.3 | 11.5 | 8.5 | 30.1 |
| cLPA C18:1 | 0.9972 | 1 | 4-533 | 1.41 | 10.7 | 11.2 | 16.4 | 26.0 | 12.9 | 12.5 |
| LPA C20:4 | 0.9914 | 4 | 42-533 | 1.81 | 7.9 | 13.3 | 33.9 | 26.8 | 17.9 | 23.9 |
| LPA C16:0 | 0.9931 | 1 | 4-533 | 1.88 | 2.9 | 14.4 | 7.7 | 26.2 | 11.6 | 25.3 |
| LPA C18:0 | 0.9899 | 1 | 4-533 | 2.25 | 26.7 | 12.2 | 44.7 | 26.8 | 12.9 | 34.6 |
| Sphingoids | ||||||||||
| Sph C18:1 | 0.9942 | 1 | 4-533 | 0.13 | 0.3 | 4.8 | 4.9 | 7.5 | 8.2 | 11 |
| Spha C18:0 | 0.9966 | 1 | 4-533 | 0.28 | 21 | 3.3 | 18.8 | 13.6 | 9.8 | 16.7 |
| S1P C18:1 | 0.997 | 1 | 4-533 | 1.52 | 4.9 | 9.3 | 24.0 | 8.4 | 16.2 | 21.0 |
| Nitro fatty acids | ||||||||||
| NO2-OA | 0.9995 | 0.09 | 0.3-173 | 0.04 | 9.9 | 5.7 | 9.1 | 9.2 | 6.0 | 10.4 |
| NO2-LA | 0.9993 | 0.09 | 0.3-173 | 0.04 | 16.3 | 8.5 | 9.5 | 14.9 | 17.5 | 20.7 |
| Prostaglandins | ||||||||||
| 2,3-Dinor-11b-PGF2α | 0.9998 | 0.09 | 0.3-173 | 0.21 | 20.8 | 8.3 | 9.8 | 13.8 | 6.6 | 5.5 |
| PGE3 | 0.9989 | 0.09 | 0.3-173 | 0.26 | 12.2 | 1.6 | 18.9 | 7.0 | 9.2 | 12.6 |
| PGD3 | 0.9996 | 0.09 | 0.3-173 | 0.21 | 7.2 | 8.8 | 15.2 | 7.7 | 6.6 | 7.7 |
| PGF3α | 0.9998 | 0.3 | 1 -173 | 0.17 | 17.2 | 5.4 | 4.9 | 17.0 | 6.6 | 7.1 |
| PGE2 | 0.9999 | 0.09 | 0.3-173 | 0.13 | 6.3 | 2.0 | 9.6 | 11.6 | 11.2 | 11.1 |
| PGE1 | 0.9996 | 0.3 | 1 -173 | 0.10 | 1.5 | 3.4 | 6.9 | 5.6 | 5.0 | 3.9 |
| PGD2 | 0.9991 | 0.09 | 0.3-173 | 0.20 | 1.1 | 2.6 | 1.5 | 1.2 | 2.3 | 1.6 |
| PGF1α | 0.9995 | 0.09 | 0.3-173 | 0.09 | 29.8 | 13.1 | 18.8 | 89.0 | 15.3 | 12.2 |
| PGF2α | 0.9987 | 0.09 | 0.3-173 | 0.09 | 15.3 | 6.6 | 6.2 | 8.8 | 5.1 | 5.2 |
| 13,14-Dihydro- PGF2α | 0.9995 | 0.09 | 0.3-173 | 0.20 | 2.6 | 4.7 | 5.2 | 2.9 | 3.4 | 3.7 |
| PGA2 | 0.9994 | 0.09 | 0.3-173 | 0.10 | 7.6 | 0.6 | 6.1 | 6.0 | 4.0 | 4.7 |
| PGA1 | 0.9995 | 0.09 | 0.3-173 | 0.07 | 1.9 | 0.7 | 5.8 | 2.7 | 7.1 | 5.5 |
| Isoprostanes | ||||||||||
| 2,3-Dinor-8-iso-PGF2α | 0.9995 | 0.09 | 0.3-173 | 0.25 | 9.5 | 2.7 | 3.6 | 11.4 | 4.3 | 3.9 |
| 8-Iso-PGF3α | 0.9995 | 0.3 | 1 -173 | 0.16 | 3.5 | 2.1 | 2.8 | 150.0 | 4.5 | 8.0 |
| 8-Iso-15-keto-PGF2β | 0.9982 | 0.09 | 0.3-173 | 0.20 | 9.1 | 5.3 | 9.2 | 12.2 | 5.8 | 7.5 |
| 8-Iso-15-keto-PGE2 | 0.9997 | 1.3 | 5-173 | 0.55 | 38.3 | 2.7 | 6.9 | 24.9 | 2.7 | 3.6 |
| 8-Iso-15-keto-PGF2α | 0.9976 | 0.09 | 0.3-173 | 0.13 | 2.6 | 4.0 | 2.3 | 15.9 | 8.1 | 11.6 |
| iPF2α | 0.9988 | 0.09 | 0.3-173 | 0.12 | 8.5 | 4.2 | 6.8 | 5.5 | 2.7 | 4.2 |
| 8-Iso-(15R)-PGF2α | 0.9991 | 0.09 | 0.3-173 | 0.12 | 10.4 | 10.2 | 9.7 | 16.3 | 12.1 | 13.8 |
| 8-Iso-PGF1α | 0.9989 | 0.09 | 0.3-173 | 0.23 | 1.1 | 4.0 | 3.9 | 3.7 | 4.1 | 3.2 |
| 8-Iso-13,14-dihydro-PGF2α | 0.9994 | 0.3 | 1 -173 | 0.11 | 10.6 | 3.7 | 6.4 | 150.8 | 8.8 | 11.6 |
| 8-Iso-PGF2α | 0.9993 | 0.09 | 0.3-173 | 0.11 | 8.1 | 5.1 | 5.3 | 9.3 | 5.1 | 6.2 |
| 8-Iso-PGE2 | 0.9997 | 0.09 | 0.3-173 | 0.12 | 5.9 | 4.2 | 2.2 | 10.9 | 19.2 | 1.7 |
| 8-Iso-PGE1 | 0.9995 | 0.09 | 0.3-173 | 0.15 | 8.2 | 3.8 | 4.8 | 43.6 | 6.9 | 3.9 |
| 5iPF2α | 0.9998 | 0.09 | 0.3-173 | 0.10 | 5.8 | 2.5 | 5.2 | 6.2 | 5.9 | 7.2 |
| 8-Iso-PGA2 | 0.9994 | 0.09 | 0.3-173 | 0.11 | 4.9 | 1.5 | 5.8 | 5.9 | 4.1 | 4.8 |
| 8-Iso-PGA1 | 0.9993 | 0.09 | 0.3-173 | 0.42 | 27.4 | 9.3 | 5.7 | 21.7 | 7.6 | 5.9 |
| 8,12-iPF2α IV | 0.9974 | 0.09 | 0.3-173 | 0.08 | 7.0 | 4.6 | 9.9 | 4.1 | 5.2 | 16.0 |
aLPA alkyl lysophosphatidic acid, cLPA cyclic lysophosphatidic acid, iP isoprostane, LA linoleic acid, LOD limit of detection, LPA lysophosphatidic acid, OA oleic acid, PG prostaglandin, RSD relative standard deviation, RT retention time, Sph sphingosine, Spha sphinganine, S1P sphingosine 1-phosphate
aCalibration curves for all standards can be found in Fig. S3.
bRT stability was calculated over three separate batches on three separate days.