TABLE 1.
Analytical validation of SFC/QqQMS for determination of lipids
| Lipid class | Ion mode | Analyte | Internal standard | RT for each analyte | RSD (%, n = 5)a | Linear range | R2 value | Range of RTsd | Spike-and-recovery teste | ||
| (min) | RRTb | RPAc | (nM) | (min) | Analyte | Recovery (%, n = 5) | |||||
| LPC | ESI+ | LPC 18:0 | LPC 17:0 | 6.12 | 0.09 | 2.7 | 50–50,000 | 0.9998 | 6.03–6.30 | LPC 18:0 | 71.9 ± 4.9 |
| LPE | ESI+ | LPE 18:0 | LPE 17:1 | 7.45 | 0.07 | 1.73 | 50–100,000 | 0.9987 | 7.32–7.56 | LPE 18:0 | 95.9 ± 3.5 |
| LPG | ESI+ | LPG 18:0 | LPG 17:1 | 10.87 | 0.08 | 8.18 | 50–100,000 | 0.9940 | – | – | – |
| LPA | ESI− | LPA 18:0 | LPA 17:0 | 17.43 | 0.57 | 8.97 | 1,000–100,000 | 0.9942 | – | – | – |
| LPI | ESI− | LPI 18:0 | LPI 17:1 | 14.02 | 0.09 | 3.5 | 5–50,000 | 0.9966 | – | – | – |
| LPS | ESI+ | LPS 16:0 | LPS 17:1 | 15.36 | 0.07 | 2.27 | 100–100,000 | 0.9999 | – | – | – |
| PC | ESI− | PC 18:0–18:2 | PC 17:0–17:0 | 5.46 | 0.08 | 4.35 | 50–50,000 | 0.9922 | 5.33–5.51 | PC 16:0-20:4 | 77.7 ± 3.5 |
| Alkyl-acyl PC (e) | ESI+ | PC 16:0e–18:1 | LPC 17:0 | 5.40 | 0.08 | 2.44 | 5–10,000 | 0.9993 | – | – | – |
| Alkenyl-acyl PC (p) | ESI+ | PC 18:0p–20:4 | LPC 17:0 | 5.45 | 0.1 | 3.65 | 50–50,000 | 0.9977 | 5.33–5.45 | PC 18:0p-20:4 | 78.0 ± 10.4 |
| PE | ESI− | PE 18:0–20:4 | PE 17:0–17:0 | 6.32 | 0.11 | 3.09 | 10–50,000 | 0.9929 | 5.80–6.32 | PE 18:0-20:4 | 101.9 ± 17.0 |
| Alkenyl-acyl PE (p) | ESI+ | PE 18:0p–20:4 | LPE 17:1 | 6.22 | 0.09 | 9.91 | 10–100,000 | 0.9976 | 6.05–6.33 | PE 18:0p-20:4 | 95.8 ± 17.3 |
| PG | ESI− | PG 16:0–18:1 | PG 17:0–17:0 | 8.99 | 0.08 | 1.72 | 5–100,000 | 0.9975 | – | – | – |
| PA | ESI− | PA 16:0–18:1 | PA 17:0–17:0 | 12.43 | 0.82 | 4.17 | 10–100,000 | 0.9985 | – | – | – |
| PI | ESI− | PI 18:0–20:4 | PS 17:0–17:0 | 12.06 | 0.1 | 2.26 | 100–50,000 | 0.9994 | 11.84–11.98 | PI 18:0-20:4 | 103.5 ± 7.6 |
| PS | ESI− | PS 18:0–18:1 | PS 17:0–17:0 | 12.23 | 0.09 | 1.87 | 50–100,000 | 0.9959 | – | – | – |
| SM | ESI+ | SM d18:1–18:0 | SM d18:1–17:0 | 5.83 | 0.09 | 2.56 | 10–10,000 | 0.9950 | 5.74–5.92 | SM d18:1-18:0 | 64.9 ± 3.0 |
| Cer | ESI+ | Cer d18:1–18:0 | Cer d18:1–17:0 | 4.48 | 0.12 | 12.95 | 10–100,000 | 0.9977 | 4.48–4.51 | Cer d18:1-18:0 | 91.0 ± 14.2 |
| CE | ESI+ | CE 16:0 | CE 17:0 | 1.12 | 1.81 | 11.32 | 50–50,000 | 0.9994 | 1.12–1.30 | CE 16:0 | 67.2 ± 5.7 |
| MAG | ESI+ | MAG 16:0 | MAG 17:0 | 3.97 | 0.18 | 3.49 | 500–100,000 | 0.9974 | 4.02–4.15 | MAG 16:0 | 98.4 ± 1.8 |
| DAG | ESI+ | DAG 18:0–20:4 | DAG 12:0–12:0 | 3.25 | 0.14 | 8.36 | 5–10,000 | 0.9976 | 3.14–3.40 | DAG 16:0-16:0 | 86.6 ± 9.1 |
| TAG | ESI+ | TAG 54:2 | TAG 51:0 | 1.61 | 3.76 | 2.86 | 500–100,000 | 0.9992 | 0.97–1.76 | TAG 54:2 | 72.8 ± 5.0 |
| FFA | ESI− | FFA 18:0 | FFA 17:0 | 4.02 | 0.11 | 1.18 | 500–100,000 | 0.9988 | 3.97–4.40 | FFA 18:0 | 91.8 ± 7.1 |
a
Amounts per injection used for validation of quantitative repeatability were 10 pmol.
b
Relative RT (analyte/internal standard).
c
Relative peak area (analyte/internal standard).
d
Range of RTs observed by the analysis of individual lipid molecular species in the all WHHLMI rabbit plasma samples in this study.
e
Representative WHHLMI rabbit plasma (placebo group) were used to spike-and-recovery test samples.