Abstract
A highly sensitive and simple LC-MS/MS method after one-step protein precipitation
was developed and validated for determination of pidotimod (CAS 121808-62-6) in human
plasma using dextrophan (CAS 125-73-5) as internal standard (IS). Pidotimod and IS
were separated on a YMC-ODS-AQ C18 column using 0.5% formic acid and methanol as a mobile phase at a flow rate of 0.3 mL/min.
Detection was performed on positive ion mode of the transitions at 245.0→134.0 for
pidotimod and 258.1→157.0 for IS by selected reaction monitoring (SRM). The assay
exhibited a linear range of 0.05–10.0 µg/mL. The lower limit of quantification were
0.05 µg/mL. Validation results indicated that the accuracy as determined from quality
control samples was in the range of − 4.00–6.48%. Intra-day and inter-day precision
was ≤ 8.35% and ≤ 8.00%, respectively. The developed method was successfully applied
to a bioequivalence study in 20 healthy Chinese volunteers following a single oral
dose of 800 mg pidotimod. The simple, inexpensive protein precipitation and high-throughput
method makes it a suitable and valuable tool in the investigation of the clinical
pharmacokinetics and bioequivalence.
Key words
pidotimod - CAS 121808-62-6 - LC-MS/MS - bioequivalence - pharmacokinetics
