Simultaneously determining the contents of combretastatin A4 myristate and its active metabolite combretastatin A4 by HPLC / 国际药学研究杂志

ABSTRACT

Objective To develop a high performance liquid chromatography (HPLC)method for simultaneously determining the contents of combretastatin A4 myristate(CA4-14)and its active metabolite combretastatin A4 (CA4) in rat plasma. Methods 1- Naphthol was used as internal standard. The analysis was performed on a Agilent Eclipse plus C18(4.6 mm × 250 mm, 5 μm) with gradient elution by using the mobile phase of acetonitrile (A)-0.4% formic acid solution(B). The column temperature was 30°C and the flow rate was 1.0 ml/min. The detection wavelength was 295 nm and the injection volume was 20 μl. Results The calibration curves were linear within the range of 0.2-100 μg/ml (r2=0.9980) for CA4-14, and 0.05-50 μg/ml(r2=0.9999)for CA4, while the quantification limits of CA4-14 and CA4 were 0.2 μg/ml and 0.01 μg/ml, respectively. The recoveries of CA4-14 and CA4 were 93.79% (RSD= 1.94%) and 96.81% (RSD=3.11%). Conclusion The method is convenient, fast, sensitive, with good precision, and can be used to determine the contents of CA4-14 and C A4 in rat plasma.