Simultaneous Determination of Seven Components in Xiaopiling Granule by HPLC Wavelength Switching Combined Gradient Elution Method

OBJECTIVE To develop an HPLC wavelength switching combined gradient elution method for determination of seven components in Xiaopiling granule simultaneously. METHODS The determination was performed on Venusil MP-C₁₈ column with mobile phase consisted of acetonitrile(A)-1% glacial acetic acid solution (B) at the flow rate of 0.9 mL·min^-1, gradient elution and the injection volum was 10 μL. The column temperature was 30 ℃. RESULTS The linear ranges of vitexin-4”-O-glucoside, vitexin-2”-o-rhamnoside, vitexin, hyperoside, ononin, calycosin and formononetin were 2.56- 51.20 µg·mL^-1, 14.87-297.40 µg·mL^-1, 2.14-42.80 µg·mL^-1, 3.16-63.20 µg·mL^-1, 3.80-76.00 µg·mL^-1, 2.14-42.80 µg·mL^-1, 4.81-96.20 µg·mL^-1, respectively(r≥0.999 1); the average recoveries were 97.0%-100.0% and the corresponding RSD was 0.55%-1.67%, respectively. The precision and the repeatability were good. Test solution was stable at room temperature in 12 h. CONCLUSION The method is simple with good precision, stability and reproducibility, and it can be used for the simultaneous determination of 7 components in Xiaopiling granule.