I don't exactly understand your question. For quantifying a compound against standard using HPLC you need to obtain a standard curve. That is made by preparing solutions of the standard at different concentrations from a stock solution (high concentrated). After that you graph the area or height of the peak from the chromatogram against each concentration value. I hope this is helpful for you.
Dear Khushboo Nawal,
if I understand your question you need to determine bisphenol A in water samples by using an external standard.
If it is the case, you have to construct a calibration curve with the reference standard, covering the concentration you believe bisphenol is in water samples.
After, you have to prepare the water sample and analyze it using the developed method. According to the peak area obtained you can determine the concentration of bisphenol A.
Assuming, your concentration is on the standard curve;
Result = (rU/rS) × (CS/CU)
rU = Peak response of Bisphenol from the Sample solution
rS = Peak response of Bisphenol from the Standard solution
CS = Concentration of in the Bisphenol Standard solution (ug/mL)
CU = Concentration of Bisphenol in the Sample solution (ug/mL)
I have carried out same research project as you questioned....but using LC-MS/MS.
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