Determination of Ferulic acid by HPLC
1) Reagents
a. Nanopure water b. Methanol (HPLC)
c.Acetic acid
2) Apparatus
HPLC (Waters) Analytical balance (1/10000) Ultrasonic cleaner
0.45μm PTFE filter 50ml volumetric flask Screw cap vials
3) HPLC condition
Column: SymmetryR C18 5μm , 3.9×150mm
Detector: UV 320nm
Mobile Phase: A=2%Acetic acid B= Methanol
A/B=65/35(isocratic)
Flow Rate: 1.0ml/min
Injection: 10μl
4) Procedure
Samples Preparation
Accurately weigh 60mg extraction into 25ml volumetric flask. Add Methanol :Acetic acid(V/V=95:5) 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 0.2mg/ml in water) using a commercial reference standard of Ferulic acid，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 Ferulic acid on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the reference standard solution. Calculate the weight percent.
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