Content Determination of 6 Flavonoids in Epimedium brevicornu from Shenqi Yanshen Granules Based on HPLC-QAMS / 中国药房

ABSTRACT

OBJECTIVE： To establish a HPLC method for simultaneous determination of 6 flavonoids in Epimedium brevicornu from Shenqi yanshen granules， such as epimedin A1， epimedin A， epimedin B， epimedin C， icariin and baohuoside Ⅰ. METHODS： HPLC method was adopted. The determination was performed on Waters Symmetry C18 column with mobile phase consisted of acetonitrile-water （gradient elution） at the flow rate of 1.0 mL/min； the column temperature was 25 ℃， and detection wavelength was 270 nm. The sample size was 10 μL. Relative correction factors （fk/s） of each component to icariin （reference substance） were established by multi-point correction method and slope correction method on the basis of external standard method to calculate the contents of each component. The contents of 6 flavonoids in 4 batches of Shenqi yanshen granules determined by HPLC external standard method were compared with by multi-point correction method and slope correction method. Feasibility and accuracy of quantitative analysis of multi-components by single-marker （QAMS） were validated. RESULTS： The linear range of epimedin A1， epimedin A， epimedin B， epimedin C， icariin and baohuoside Ⅰ were 2.03-50.80 μg/mL （r＝0.999 5）， 4.34-108.60 μg/mL （r＝0.999 5）， 2.26-56.40 μg/mL （r＝0.999 5）， 4.14-103.60 μg/mL （r＝0.999 5）， 4.24-106.00 μg/mL （r＝0.999 5）， 1.78-44.60 μg/mL （r＝0.999 5）， respectively， the limits of detection were 65.80， 71.49， 74.26， 68.79， 70.56， 86.09 ng/mL， respectively； the limits of quantification were 196.62， 213.63， 223.72， 208.46， 215.96， 255.88 ng/mL， respectively； RSDs of precision， stability （24 h）， reproducibility tests were less than 2％ （n＝6）， respectively. The average recoveries were 96.03％-99.04％ （RSDs were 0.65％-1.04％， n＝6）. By multi-point correction method， fk/s of epimedin A1， epimedin A， epimedin B， epimedin C and baohuoside Ⅰ were 0.837， 0.818， 0.845， 0.831， 1.387， respectively； by slope correction method， fk/s of them were 0.835， 0.815， 0.851， 0.829， 1.419， respectively. There was no significant difference in content determination results between two correction methods of QAMS and external standard （P＞0.05）. CONCLUSIONS： Established HPLC- QAMS method is accurate and suitable for the quality control of epimedin A1， epimedin A， epimedin B， epimedin C， icariin and baohuoside Ⅰ in E. brevicornu from Shenqi yanshen granules.