Kavain by HPLC
Determination of Kavain by HPLC
1) Reagents
a. Methanol (HPLC) b. Nanopure water c.Acetronitrile(HPLC) d. Phoaphate acid（C.R）
2) Apparatus
HPLC (Shimadzu) Analytical balance (1/10000) Ultrasonic cleaner
0.45μm PTFE filter 50ml volumetric flask Screw cap vials
3) HPLC condition
Column: Shim-Pack VP-ODS C18 5 m 4.6150mm
Detector: UV 256nm
Mobile Phase: A:0.1% Phoaphate acid B: MeOH: Acetronitrile=1:1
A:B=45:55
Flow Rate: 1.0ml/min
Injection: 10μl
4) Procedure
Samples Preparation
Accurately weigh 20mg extraction into 50ml volumetric flask. Add 5% MeOH 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 mixed standard solution (approximately 0.6mg/ml in 5% MeOH) using a commercial reference standard of Kavain，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 Kavain on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the reference standard solution. Calculate the weight percent.
1) Reagents
a. Methanol (HPLC) b. Nanopure water c.Acetronitrile(HPLC) d. Phoaphate acid（C.R）
2) Apparatus
HPLC (Shimadzu) Analytical balance (1/10000) Ultrasonic cleaner
0.45μm PTFE filter 50ml volumetric flask Screw cap vials
3) HPLC condition
Column: Shim-Pack VP-ODS C18 5 m 4.6150mm
Detector: UV 256nm
Mobile Phase: A:0.1% Phoaphate acid B: MeOH: Acetronitrile=1:1
A:B=45:55
Flow Rate: 1.0ml/min
Injection: 10μl
4) Procedure
Samples Preparation
Accurately weigh 20mg extraction into 50ml volumetric flask. Add 5% MeOH 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 mixed standard solution (approximately 0.6mg/ml in 5% MeOH) using a commercial reference standard of Kavain，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 Kavain on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the reference standard solution. Calculate the weight percent.