Determination of Osthol by HPLC
1) Reagents
a. Acetronitrile (HPLC) b. Nanopure water
2) Apparatus
HPLC (shimadzu) 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 320nm
Mobile Phase: A= Water B= Acetronitrile
Flow Rate:1.0ml/min
Injection: 10μl
Gradient:
Time(min) %A %B
0.01 77 23
50 35 65
55 35 65
57 77 23
60 77 23
4) Procedure
Samples Preparation
Accurately weigh 30mg extraction into 50ml volumetric flask. Add (Water: Acetronitrile
=1:1V/V) 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 0.3mg/ml in Water: Acetronitrile
=1:1V/V) using a standard of Osthol，Sonicate for about 10 minutes and transfer to screw-cap vials and can be stored in the refrigeration.
5) Calculation
Determine the peak areas of the Osthol on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the standard solution. Calculate the weight percent.
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