Program | Hplc

Most modern HPLC systems use chromatography data systems (CDS). You will fill in a table like this:

[e.g., Agilent 1260 Infinity II, Waters Alliance e2695] Column: [e.g., C18, 150 x 4.6 mm, 5 µm] Detection: [e.g., UV-Vis at 254 nm / PDA / Fluorescence] hplc program

Assuming a mixture of Paracetamol (polar) and Ibuprofen (non-polar). Most modern HPLC systems use chromatography data systems

| Problem | Check This Program Setting | Fix | |---------|----------------------------|------| | No peaks | Detector wavelength | Confirm λmax of analyte | | Negative peaks | Reference wavelength | Set ref to offline or 600 nm | | Retention time creep | Gradient start time | Ensure re-equilibration step exists | | High pressure | Flow rate gradient | Look for programmed flow >1.5 mL/min | | Ghost peaks | Needle wash program | Increase wash volume/time | | Baseline spikes | Lamp program | Is lamp on "warm-up" mode? Switch to "run" | | Poor peak shape | Column temperature | Is oven actually on? Check program activation | Switch to "run" | | Poor peak shape