Table 2.
MRM conditions in UPLC-QqQ-MS/MS analysis and method validation for quantitative determination.
| Analyte | MRM | Collision voltage (eV) | Calibration curve | Sensitivity (ng/mL) | (RSD, %) | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Repeat-ability (n = 6) | Precision (n = 6) | Spike Recovery (n = 3) | Stability (72 h, n = 7) | |||||||||||
| Range (ng/mL) | Equation | R2 | LOD | LOQ | Intra-day | Inter-day | Low | Middle | High | |||||
| Gallic acid | 169.0→125.0 | 11 | 80–4000 | y = 79.28x−14.86 | 0.9968 | 17.94 | 56.96 | 2.84 | 2.17 | 5.79 | 106.88 (0.87) | 104.22 (3.85) | 112.40 (2.18) | 7.95 |
| Proanthocyanidin B1 | 577.1→407.0 | 23 | 80–2,000 | y = 68.49x−1084.17 | 0.9959 | 12.50 | 15.66 | 4.96 | 1.13 | 3.56 | 113.48 (7.66) | 105.21 (0.42) | 102.97 (5.04) | 6.07 |
| Proanthocyanidin B2 | 577.1→407.0 | 23 | 80–2,000 | y = 78.28x−1088.89 | 0.9979 | 8.43 | 11.01 | 7.42 | 4.29 | 2.52 | 99.74 (7.16) | 111.41 (8.21) | 100.49 (11.86) | 9.44 |
| Epicatechin | 289.1→245.1 | 7 | 40–1,000 | y = 38.44x−203.28 | 0.9916 | 4.08 | 38.96 | 6.25 | 5.59 | 11.74 | 89.75 (11.98) | 106.90 (3.54) | 96.42 (6.38) | 7.30 |
| Epcatechini-3-gallate | 441.1→169.0 | 15 | 20–400 | y = 125.61x−300.88 | 0.9916 | 4.81 | 7.61 | 2.00 | 2.79 | 3.56 | 100.23 (1.02) | 93.55 (1.39) | 93.78 (1.46) | 3.09 |
| trans-THSG | 405.0→243.0 | 15 | 160–8,000 | y = 288.41x−11468.51 | 0.9951 | 2.14 | 3.37 | 1.49 | 1.29 | 3.48 | 96.23 (5.62) | 98.23 (7.38) | 93.92 (4.80) | 5.90 |
| Emodin-8-O-β-D-glucoside | 431.1→269.1 | 27 | 40–2,000 | y = 556.32x−6573.43 | 0.9943 | 1.88 | 2.30 | 1.05 | 1.09 | 11.42 | 88.98 (4.00) | 94.85 (4.66) | 94.90 (6.70) | 5.40 |
| Physcion-8-O-β-D-glucoside | 445.1→283.1 | 7 | 40–2,000 | y = 89.55x−643.74 | 0.9953 | 4.57 | 6.85 | 2.59 | 3.09 | 8.03 | 93.32 (8.90) | 98.60 (8.00) | 92.25 (2.49) | 10.34 |
| Emodin | 269.0→225.0 | 25 | 40–2,000 | y = 758.58x−4801.43 | 0.9992 | 2.53 | 3.28 | 0.98 | 1.06 | 1.07 | 108.12 (1.23) | 105.40 (1.58) | 107.03 (0.58) | 5.43 |
| Physcion | 283.0→240.0 | 20 | 40–2,000 | y = 200.82x−410.28 | 0.9965 | 1.69 | 5.18 | 1.55 | 2.74 | 1.01 | 109.14 (3.06) | 108.77 (2.00) | 106.25 (1.58) | 6.09 |