Background: Chromatographic methods for determination of insulin degludec in rabbit
plasma by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry were developed.
Methods: Analytes were eluted from Waters ACQUITY UPLC® Peptide BEH C18 (2.1×50mm, 300Å)
column with a mobile phase of water containing 0.1% formic acid (A) and acetonitrile containing 0.1%
formic acid (B). Quantitation of insulin degludec was performed using 1222.06 > 641.24 m/z on Multiple-
Reaction Monitoring (MRM) mode.
Results: Good linearity was observed in the concentration range of 500-50000 ng/mL (r >0.99), and the
lower limit of quantification was 500ng/mL. The within-run and between-run precision (expressed as
relative standard deviation, RSD) of insulin degludec were ≤ 14.16% and ≤ 13.64% respectively, and
the accuracy was within 94.37-96.35%. The recovery and matrix effects were both within acceptable
Conclusions: This method was successfully applied for the pharmacokinetic study of insulin degludec
in rabbit after subcutaneous administration.