Dear researchers may i know, how to calculate the concentration of curcumin,DMC and BDMC from its peak area in HPLC , PDA detector? I am using isochractic flow of acetonitrile and 0.1 % O-Phosphoric acid in the ratio of 60:40 respectively.
Hi, for doing quantification by HPLC-DAD, I think at first you should run the authentic standards of C, DMC and BDMC, and compare the UV spectra of your target peaks to the UV spectra of the standards, after that you should check the purity of the target peaks; if all are OK, it means that you have good selectivity. Then you should develop calibration standards from the 3 authentic standards in the range +/- 20-30% of the expected concentrations, the concentrations of your target compounds can be calculated from the each of calibration curves.
If you want just to compare the (relatively) concentrations of C, DMC, BDMC in your sample, you can do by compare their peak areas (normalization); Relative concentration of C = [ area C / (Area C+DMC+BDMC) x !00 %; Analog for DMC and BDMC
1. Internal standard (As , Cs ) analyte with known concentration (Ax , Cx )
As: peak area of standard
Cs: concentration of analyte
analyte with unknown concentration
Ax: peak area of analyte
Cx: concentration of standard
2. Standard Curve
Calibration curve of concentration of analyte (CA) versus response (RA) for external standardization
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