Winnonlin professional version 4

Male ICR mice (18–20 g, n = 3 in each group) were randomly assigned to treatment groups. After 12-h of fasting, baseline blood was collected into a tube containing ethylenediaminetetraacetic acid (EDTA). Animals were then treated orally with compound A57 or APC (p.o.: 20 mg/kg, in DMSO/0.5% HPMC 5/95, v/v) and intravenous injected the selected compounds (i.v.: 2 mg/kg, in DMSO/EtOH/PEG300/NaCl 5/5/40/50, v/v/v/v). Subsequently, blood samples per mouse were collected at 0.25, 0.5, 1, 2, 4, 8, 24 h for p.o.-treated groups and 0.05, 0.25, 0.75, 2, 4, 6, 8, 24 h for i.v.-treated groups. All samples were centrifuged at 12,000 rpm for 3 min, and the plasma was harvested. Aliquots of plasma samples were stored at −80 °C until analysis. The selected compounds' serum concentrations were determined by liquid chromatography/tandem mass spectrometry (LC–MS–MS). Pharmacokinetic parameters were determined from the selected compounds' serum concentrations by noncompartmental methods using WinNonLin professional version 4.1 (Pharsight Corp., Mountain View, CA).

XG-102 concentrations in plasma samples were determined by a validated high-performance liquid chromatography, tandem mass spectrometry assay with a lower limit of quantification (LLOQ) of 10 ng/mL; this analytical method is considered precise, accurate, and specific with the following characteristics: intra-precision (1.4–18.8 acceptable because it is concerning the LLOQ), inter-precision (5.7–16.8 acceptable because it is concerning the LLOQ); intra-accuracy (91–100%), inter-accuracy (100–113%); and specificity (no interference <5%). Quantification of XG-102 in the infusate was performed using a validated HPLC-ultraviolet assay with a LLOQ of 1 μg/mL is also defined by the following characteristics: intra-precision (0.7–3.7), inter-precision (3.3–6.3); intra-accuracy (96.6–107%), inter-accuracy (95.4–113%); and specificity (no interference <5%).
The pharmacokinetic analysis was conducted by BCRU using WinNonlin Professional Version 4.1 (Pharsight Corporation, Mountain View, CA).
Pharmacokinetic parameters were determined from the plasma concentrations of XG-102 using noncompartmental procedures. The PK parameters determined are presented in Table 1.
AUC_0–last (area under the curve) was calculated using the linear log-trapezoidal method.