Research Article Zangarini, Rajan, Danilenko, Berry, Traversa & Veal
Table 3. LLOQ of pegcantratinib in human tumor homogenate. Actual concentration (ng/ml)
2.5 – – – –
Mean (n = 6) SD
Accuracy (%) Precision (%)
Found concentration (ng/ml) 2.22 2.57 2.58 2.41 2.59 2.47 0.16 99
6.0
samples stored at -20°C for 3 months. The pegcantra- tinib QC samples were analyzed against a calibration curve, obtained from freshly spiked calibration stan- dards, and the obtained concentrations were compared with the nominal concentrations. The mean concen- tration at QC concentration was required to be within ±15% of the nominal concentration [12,13].
Analysis of clinical samples Biopsies weighing 5–20mg were obtained from cyl- indroma skin tumors of two CYLD carrier patients enrolled in a Phase Ib open label clinical trial (ISRCTN: 75715723) that received daily topical doses of pegcantratinib (0.5% w/w) for 4 weeks. Biopsies were excised at the end of treatment as described above.
Accuracy (%) 89
103 103 96
104 – – – –
Results Optimization of HPLC–MS/MS parameters & conditions Using an ESI source in positive ion mode coupled with in-source CID, pegcantratinib and IS generated an abundant fragment ion after applying an optimized DP. As seen previously, the mass spectrum of pegcantratinib at a nondissociating voltage of 20 V (Figure 2A) com- pared with a high DP of 186 V (Figure 2B), showed the disappearance of the characteristic broad profile associ- ated with PEGylated molecules. This indicates that the polymeric species was dissociated and an abundant new ion species at m/z 419.0 was generated (Figure 1). The PEG disassociation was obtained for both the analyte and the IS. The newly obtained ion (m/z 419.0) passed
Figure 4. Calibration curve showing linearity over six concentrations of pegcantratinib (range: 2.5–250 ng/ml) from three separate experiments.
28421 Bioanalysis (2017) 9(3) future science group
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