Curcumin of Curcumae by HPLC
Determination the Curcumin of Curcumae by HPLC
1) Reagents
a. Nanopure water b. Methanol (HPLC)
c. .Acetic acid（A.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: Kromasil C18 5 m 4.6200mm
Detector: UV 425nm
Mobile Phase: A:=2%.Acetic acid B: Methanol
Gradient:
Time(min) %A %B
0.01 52 48
14 45 55
16 45 55
17 52 48
20 52 48
Flow Rate: 0.9ml/min
Injection: 10μl
4) Procedure
Samples Preparation
Accurately weigh 20mg extraction into 100ml volumetric flask. Add Methanol to volumetric flask and sonicate to mix, cool to room temperature and add Methanol to volume,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 0.10mg/ml in methanol using a standard of Curcumin，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 Curcumin on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the standard solution. Calculate the weight percent .
1) Reagents
a. Nanopure water b. Methanol (HPLC)
c. .Acetic acid（A.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: Kromasil C18 5 m 4.6200mm
Detector: UV 425nm
Mobile Phase: A:=2%.Acetic acid B: Methanol
Gradient:
Time(min) %A %B
0.01 52 48
14 45 55
16 45 55
17 52 48
20 52 48
Flow Rate: 0.9ml/min
Injection: 10μl
4) Procedure
Samples Preparation
Accurately weigh 20mg extraction into 100ml volumetric flask. Add Methanol to volumetric flask and sonicate to mix, cool to room temperature and add Methanol to volume,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 0.10mg/ml in methanol using a standard of Curcumin，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 Curcumin on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the standard solution. Calculate the weight percent .