Acyclovir

As described previously [23 (link), 35 (link)], vitamin C concentrations in WT or Svct1 KO mice plasma were determined using liquid chromatography-photodiode array (LC-PDA) analysis with an ACQUITY UPLC^® PDA Detector (Waters, Milford, MA, USA) coupled with an ACQUITY UPLC System (Waters). After deproteinization with equal volumes of 10% (w/v) metaphosphate solution containing 1 mM EDTA, each sample was centrifuged at 20,000 × g at 4°C for 10 min,the obtained supernatant was diluted with equal volumes of 25 mM phosphate buffer (pH 2.1) containing 60 μM acyclovir (FUJIFILM Wako Pure Chemical) as an internal control and then separated on a CAPCELL PAK ADME S3 column (maintained at 50°C; 3 μm, 2.1 × 100 mm; Osaka Soda, Osaka, Japan) with a gradient mobile phase (0–4 min: 2% B; 4–5 min: 2–98% B; 5–9 min: 98% B; 9–10 min: 98–2% B; 10–12 min: 2% B) of 25 mM phosphate buffer (pH 2.1) (A) and methanol (B) at a flow rate of 300 μL/min. Vitamin C and acyclovir were measured at 243 nm in the PDA spectrum. Calibration curves for the analyte were generated using a series of murine plasma spiked with L-ascorbic acid (the reduced form of vitamin C) standard solutions. Peaks were analyzed using MassLynx NT software v4.1 (Waters).