|Determination of Epimedium by HPLC
a. Nanopure water b. Acetronitrile(HPLC) c.Acetic acid (A.R)
HPLC (Shimadzu) Analytical balance (1/10000) Ultrasonic cleaner
0.45μm PTFE filter 50ml volumetric flask Screw cap vials
3) HPLC condition
Column: Kromasil C18 5 m 4.6200mm
Detector: UV 270nm
Mobile Phase: A=Water: HAC (50:1) B=Acetronitrile
Flow Rate: 0.7ml/min
Accurately weigh 20mg extraction into 50ml volumetric flask. Add Methanol to volume and sonicate to mix, filter through a 0.45μm PTFE filter into a screw cap vial and proceed with the analysis.
Standard preparation /calibration
Prepare a standard solution (approximately 0.1mg/ml in Methanol) using a standard of Epimedium，Sonicate for about 10 minutes and transfer to screw-cap vials and can be stored in the refrigeration.
Determine the peak areas of the Epimedium on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the standard solution. Calculate the weight percent.