Dilute your HPLC standards to the desired concentrations using a compatible solvent (generally bacteriostatic water for all water soluble compounds)
Loading the Autosampler:
Transfer the prepared standards into HPLC vials with tight-sealing caps to prevent contamination or evaporation. Label the vials clearly for easy identification. Place the vials in the autosampler tray according to the sample sequence you have programmed.
Programming the Autosampler:
Input the details of the standards, such as vial position, volume to inject, and run order, into the HPLC software. Ensure the autosampler is programmed to perform any necessary rinsing steps to avoid cross-contamination.
Running the Standards:
Begin the calibration run, allowing the autosampler to inject the standards into the HPLC system automatically. Monitor the chromatogram to confirm the peaks align with the known retention times and concentrations of the standards.
Data Analysis:
Use the chromatograms generated from the standards to create a calibration curve. Compare the peak areas or heights to determine the concentration of analytes in your unknown samples.