I am studying the in vitro release study of a drug nanoparticle. When I inject the standard drug solution to HPLC to make a calibration curve, it shows different areas of the peak of the same amount of std drug solution added.
It might be because of contamination in solvents, tubing, injector, or even column. You might want to check the stability of that drug as well. Good luck!!
Make sure whether your substance is dissolved completely. What's the substance? There may be a method describing an appropriate preparation procedure for the HPLC analysis. Take down the title of this substance
You might have a problem with either the sample preparation process or the sample loop is contaminated from a previous analysis.
Try reviewing the method of sample preparation and see if you did anything that could have compromized the purity of your drug.
If not from the preparation process, it could be that your sample loop has traces/contaminants from previously analyzed samples. You could solve this by flashing/rinsing the sample loop with an appropriate solvent depending on the solubility of the previously analyzed sample suspected to be the contaminant several times.