Table 2 Precision and accuracy data for the quantification of six nucleosides in mRNA (n = 5)
Analytes | Concentration levels (mean ± SD, pg/mL) | RSD (%) | RE (%) | ||
|---|---|---|---|---|---|
Added | Measured | Intra-day | Inter-day | Accuracy | |
Adenosine (A) | 40 | 40.8 ± 1.9 | 6.5 | 4.3 | 2.0 |
120 | 133.3 ± 3.1 | 3.9 | 2.0 | 11.1 | |
1600 | 1792.7 ± 33.7 | 3.6 | 1.4 | 12.0 | |
16,000 | 15,120.0 ± 270.4 | 4.0 | 1.0 | − 5.5 | |
Uridine (U) | 2000 | 1919.3 ± 96.0 | 6.5 | 4.7 | − 4.0 |
6000 | 6101.3 ± 220.7 | 7.4 | 2.5 | 1.7 | |
80,000 | 83,680.0 ± 1235.9 | 2.6 | 1.2 | 4.6 | |
800,000 | 710,133.3 ± 10,405.1 | 2.1 | 1.3 | − 11.2 | |
Cytidine (C) | 40 | 36.7 ± 2.3 | 0.5 | 6.8 | − 8.3 |
120 | 117.3 ± 5.4 | 4.8 | 4.5 | − 2.3 | |
1600 | 1672.7 ± 26.0 | 1.5 | 1.6 | 4.5 | |
16,000 | 15,953.3 ± 247.5 | 1.2 | 1.6 | − 0.3 | |
Guanosine (G) | 200 | 195.0 ± 12.0 | 12.7 | 4.1 | − 2.5 |
600 | 630.4 ± 21.8 | 6.4 | 2.7 | 5.1 | |
8000 | 8472.0 ± 280.3 | 4.3 | 3.1 | 5.9 | |
80,000 | 78,180.0 ± 1894.1 | 4.5 | 1.9 | − 2.3 | |
N6-Methyladenosine (m6A) | 40 | 36.2 ± 2.0 | 13.8 | 1.9 | − 9.5 |
120 | 131.2 ± 6.9 | 2.4 | 5.6 | 9.3 | |
1600 | 1733.3 ± 96.0 | 7.2 | 5.2 | 8.3 | |
16,000 | 15,806.1 ± 196.9 | 1.2 | 1.2 | − 1.2 | |
N5-Methylcytidine (m5C) | 40 | 38.4 ± 1.6 | 2.1 | 4.3 | − 4.0 |
120 | 122.3 ± 3.3 | 4.2 | 2.3 | 1.9 | |
1600 | 1697.3 ± 39.9 | 6.0 | 0.6 | 6.1 | |
16,000 | 15,726.7 ± 286.5 | 3.3 | 1.4 | − 1.7 | |