No peaks or small peaks

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Probable cause		Suggested actions
1	NOTE: The eluents should be miscible and should not form salt precipitates over the range of the entire gradient. The eluents should also be properly degassed. NOTE: Due to its miscibility with water and most organic solvents, isopropanol is often the choice when removing air bubbles from an HPLC system.	Ensure that the method settings are properly set and the gradient system is delivering the correct gradient composition. Check if the mobile phase reservoirs are empty or if the filter frits are blocked. Check for air bubbles in the system. If necessary, purge the pump and system with a viscous mobile phase to remove any air bubbles. Check the system for any leaks. Check for leaking pump seals. If necessary, replace pump seals. Check for sticking check valves. If necessary, sonicate or replace check valves.
2	NOTE: The eluents should be miscible and should not form salt precipitates over the range of the entire gradient. The eluents should also be properly degassed.	Ensure mobile phase and column compatibility. Ensure solvent lines are associated with the desired mobile phases. Use gradient elution that ends at a relatively high organic composition, such as 95 % acetonitrile, to ensure that all components are eluted during the analysis. Increase the starting organic composition, use a higher flow rate, or increase the overall strength of the eluent system to get the components to elute quicker from the column.
3	Aging detector lamp	Check if the on-time hours of the detector lamp have exceeded the expected lifetime. Run detector lamp intensity test to check the condition of the lamp. Replace the lamp if necessary.
4	Improper method settings	Check the detector method settings , such as wavelength and data rate, and optimize them if needed. Check temperature settings in the column compartment.
5	Small detector cell	Make sure the detector cell has the same path length as the one used for the expected chromatogram.
6	Strong retention	Try a less hydrophobic stationary phase for the column. Use columns with PEEK hardware and implement the use of InfinityLab Deactivator Additive or similar when working with metal-binding analytes.
7	NOTE: If the vial is full, the sampling volume could be smaller than expected due to vacuum effects when drawing the sample. All some space in the vial by not filling it passed the vial’s shoulder.	Ensure that the vials are not filled to the top.

21-Jun-2025