Table 2 Summary of the validation results of the developed LC–MS/MS method in MRM mode
Validation parameter | K9[Poy]K14[Poy]-heavy peptide | K9[Poy]K14[Ac]-heavy peptide |
|---|---|---|
LLOD | 0.106 fmol/µL | 0.204 fmol/µL |
LLOQ | 0.5 fmol/µL | 0.5 fmol/µL |
ULOQ | 2500 fmol/µL | 2500 fmol/µL |
Accuracy | 93.2–111% | 88.9–112.5% |
Precision | 0.44–5.48% (%CV) | 0.83–4.85% (%CV) |
Linearity | R2 = 0.9998 | R2 = 0.9997 |
Linear range | 0.5–2500 fmol/µL | 0.5–2500 fmol/µL |
Sensitivity | 0.5 fmol/µL | 0.5 fmol/µL |
Specificity | Unique MRM transitions for K9[Poy]K14[Poy]-heavy peptide were measured | Unique MRM transitions for K9[Poy]K14[Poy]-heavy peptide were measured. MRM transitions that overlap with the isobaric K9[Ac]K14[Poy]-heavy peptide were excluded |
Selectivity | No interfering signals in the region of the analyte elution | No interfering signals in the region of the analyte elution |