|Determination of Fufang of Herbal Drugs by HPLC
a. Acetronitrile (HPLC) b. Phosphate acid(A.R) c. Nanopure water
HPLC (Beckman) Analytical balance (1/10000) Ultrasonic cleaner
0.45μm PTFE filter 25ml volumetric flask Screw cap vials
3) HPLC condition
Column: Kromasil C18 5 m 4.6200mm
Detector: UV 274nm
Mobile Phase: A= Water B= Acetronitrile
Time %A %B
0.01 95 5
30 50 50
40 20 80
50 10 90
60 10 90
62 95 5
66 95 5
Flow Rate: 1.3ml/min
Accurately weigh 120mg extraction into 25ml 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.
Reference preparation /calibration
Prepare an reference solution (approximately 0.4mg/ml in MeOH) using a commercial reference of Fufang of Herbal Drugs，Sonicate for about 10 minutes and transfer to screw-cap vials and can be stored in the refrigeration.
Compare the peak areas of the Fufang of Herbal Drugs on the sample chromatogram and the corresponding peaks in the chromatogram for the reference solution.