Purpose: To develop a simple, accurate, and
precise high performance chromatography (HPLC) method
with spectrophotometric detection for the determination
of doxorubicin hydrochloride in rat plasma.
Methods:
Doxorubicin hydrochloride and daunorubicin hydrochloride
(internal standard, IS) were separated on a C18
reversed-phase HPLC column. Following protein
precipitation extraction, chromatographic separation was
accomplished with a mobile phase consisting of
acetonitrile: water at ratio of 30:70 (pH 3.0), and the
drug was detected at 233 nm using a UV detector at flow
rate of 1.0 ml/min and ambient temperature.
Results:
Linearity was obtained over the range 1.0 – 50.0 μg/ml
for doxorubicin hydrochloride with lower limit of
quantitation of 1.0 μg/ml. For each level of quality
control samples, inter- and intra-day precision (% CV)
was < 9.6 and 5.1 %, respectively. Stability of
doxorubicin hydrochloride in plasma was within the
acceptance limit (± 15 %) with no evidence of
degradation during sample processing and 30 days storage
in a deep freezer at -70 ± 5 °C. Absolutes extraction
recovery of drug from plasma was ≥ 86 %.
Conclusion: The method is highly selective and rugged for the
determination of doxorubicin hydrochloride in rat plasma
and should be suitable for conducting pharmacokinetic
studies and therapeutic drug monitoring.
Keywords:
Doxorubicin, Daunorubicin,
Validation, pharmacokinetics, rat plasma
