Bioprotocols

Troubleshooting High Performance Liquid Chromatography (from ASRG)

 

1.      Peaks 

 

Broad peaks

Ghost peaks

Negative peaks

Peak doubling

Peak fronting

Peak tailing

Spikes

 

2.      Leaks 

 

Leak at column or fittings

Leak at detector

Leak at injection valve

Leak at pump

 

 

3.      Recovery 

 

Poor sample recovery

 

4.      Sensitivity 

 

Lack of sensitivity

 

5.      Retention 

 

Changing retention times

Decreasing retention times

Increasing retention times

Retention beyond total permeation volume

 

6.      Equilibration 

 

Slow column equilibration time

Varying retention times

 

7.      Baseline 

 

Disturbance at void time

Drifting baseline

Noise

 

8. Pressure 

Decreasing pressure

Fluctuating pressure

High back pressure

Increasing pressure

 

 

HPLC Trouble Shooting (Shula Levin’s Lab)

 

Abnormal Peak Shape

 

Fluctuation in Elution Time

 

Fluctuation in Baseline

 

Noise in Baseline

 

Rise in Pressure

 

General Care to Use Packed Columns

 

Column Cleaning Procedure

 

Column Test Conditions