UPLC-MS/MS method for quantification of 3,14,19-Triacetyl
andrographolide in rat plasma: application to preclinical
pharmacokinetic studies
Abstract
Rationale: 3,14,19-Triacetyl andrographolide (ADA), a derivative of
andrographolide, possesses promising anti-inflammatory and anti-tumor
properties. However, there is currently no pharmacokinetic study of ADA.
Accurate quantification of ADA in biological matrices is crucial for
evaluating its pharmacokinetic profile and therapeutic potential. Here,
we aimed to develop a rapid and sensitive UPLC-MS/MS method for
quantifying ADA in rat plasma, facilitating preclinical pharmacokinetic
studies. Methods: An ACQUITY UPLC BEH C18 column was employed for
chromatographic separation of ADA and celecoxib (internal standard, IS)
using a gradient mobile phase of acetonitrile/water (both containing
0.1% formic acid) at a flow rate of 0.3 mL/min. Quantification was
achieved via multiple reaction monitoring (MRM) transitions of m/z
499.1→439.1 for ADA and m/z 381.9→361.9 for celecoxib. Plasma samples
underwent liquid-liquid extraction prior to analysis. The method’s
linearity, precision, accuracy, matrix effects, and recovery were
thoroughly evaluated. Additionally, we administered different doses of
ADA via two routes of administration (intragastric administration and
intravenous injection) in rats and determined its plasma pharmacokinetic
profile in vivo, including estimating its oral bioavailability. Results:
The developed method exhibited excellent linearity (R
2=0.9921) over a concentration range of 5-2000 ng/mL
for ADA in rat plasma. Both within-run and between-run precision (RSD%)
were less than 6.88%, and the accuracy ranged from 94.86% to 107.61%.
Matrix effects were found to be between 87.53% and 112.78%, with
recoveries ranging from 98.60% to 104.39%. ADA was stable under
various storage conditions in plasma. Applying this validated method to
pharmacokinetic studies in rats showed that ADA exhibited linear
pharmacokinetic characteristics in the dose range of 100-400 mg/kg, and
the oral bioavailability was 7.193%-9.169%. Conclusion: The
established UPLC-MS/MS method provides a rapid, sensitive, and reliable
tool for the quantification of ADA in rat plasma, enabling comprehensive
pharmacokinetic studies and further evaluation of its therapeutic
potential.