Ginkgo Flavone by HPLC
Determination of Ginkgo Flavone by HPLC
1) Reagents
a. Methanol(HPLC) b. Nanopure water c.Phosphoric acid(A.R)
2) Apparatus
HPLC (Shimadzu) Analytical balance (1/10000) Ultrasonic cleaner
0.45μm PTFE filter volumetric flask Screw cap vials
3) HPLC condition
Column: Kromasil C18 5m 4.6×200mm
Detector: UV 368nm
Mobile Phase: A=0.4% Phosphoric acid B=Methanol
Flow Rate: 1.1 ml/min
Injection: 10μl
Gradient:
Time(min) %A %B
0.01 60 40
22 36 64
25 60 40
30 60 40
4) Procedure
Samples Preparation
Accurately weigh 200mg extraction into 100ml flask. Add 20ml Methanol to it 、 sonicate to dissolve and mix , add 5ml 25%HCLand heated under reflux on a boiling water bath for 3hr. then cool to room temperature and transfer to the 100ml volumetric flask ,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 Quercitin,Kaempferol,Isorhamnatine ，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 Quercitin,Kaempferol,Isorhamnatine on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the standard solution. Calculate the weight percent.
1) Reagents
a. Methanol(HPLC) b. Nanopure water c.Phosphoric acid(A.R)
2) Apparatus
HPLC (Shimadzu) Analytical balance (1/10000) Ultrasonic cleaner
0.45μm PTFE filter volumetric flask Screw cap vials
3) HPLC condition
Column: Kromasil C18 5m 4.6×200mm
Detector: UV 368nm
Mobile Phase: A=0.4% Phosphoric acid B=Methanol
Flow Rate: 1.1 ml/min
Injection: 10μl
Gradient:
Time(min) %A %B
0.01 60 40
22 36 64
25 60 40
30 60 40
4) Procedure
Samples Preparation
Accurately weigh 200mg extraction into 100ml flask. Add 20ml Methanol to it 、 sonicate to dissolve and mix , add 5ml 25%HCLand heated under reflux on a boiling water bath for 3hr. then cool to room temperature and transfer to the 100ml volumetric flask ,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 Quercitin,Kaempferol,Isorhamnatine ，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 Quercitin,Kaempferol,Isorhamnatine on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the standard solution. Calculate the weight percent.