High-Performance Liquid Chromatography (HPLC) is a widely used analytical technique in the field of analytical chemistry. The HPLC instrument works by passing a sample through a column, separating its components, and detecting them using a detector. The detector typically measures the sample's absorbance at a specific wavelength, and the data generated can be used to identify and quantify the sample's components.
However, sometimes problems can arise during the HPLC analysis, which can affect the accuracy and precision of the results. In this article, we will discuss troubleshooting problems detected by smell, sight, or sound in HPLC.
Smell
A strong, unpleasant odor during an HPLC analysis can indicate several problems. One possible cause is the contamination of the mobile phase, which can lead to unwanted reactions with the sample or the column's stationary phase. Contamination can occur if the mobile phase is not properly prepared, if the solvent used is of low quality, or if there is an issue with the solvent delivery system. To address this issue, the mobile phase should be prepared correctly, and the solvent delivery system should be cleaned and maintained regularly.
Another possible cause of a strong odor during HPLC analysis is a problem with the column. Columns can become contaminated or damaged over time, leading to unwanted reactions and unpleasant odors. To address this issue, the column should be checked and replaced if necessary.
Sight
Problems detected by sight in HPLC can include changes in the baseline or peaks, changes in retention times, or changes in peak shape. One possible cause of these issues is an improperly prepared mobile phase. The mobile phase should be prepared according to the correct protocol and should be free of any contaminants.
Another possible cause of problems detected by sight in HPLC is a problem with the column. Columns can become clogged, contaminated, or damaged, leading to changes in peak shape, retention times, or baseline. To address this issue, the column should be checked, cleaned, or replaced if necessary.
Sound
Problems detected by sound in HPLC can include irregular or inconsistent flow rates, which can affect the accuracy and precision of the results. One possible cause of these issues is an improperly prepared mobile phase, which can affect the flow rate and pressure of the system. To address this issue, the mobile phase should be prepared according to the correct protocol, and the solvent delivery system should be maintained regularly.
Another possible cause of problems detected by sound in HPLC is an issue with the pump or the instrument's hardware. The pump or other hardware components may become clogged or damaged, leading to irregular flow rates or inconsistent pressure. To address this issue, the instrument should be checked, and the pump or other hardware components should be cleaned or replaced if necessary.
Troubleshooting Problems Detected by Smell, Sight or Sound in HPLC
You need to use all your senses to identify LC problems. You should get in the habit of taking a few minutes each day to expose all of your senses (except taste!) to the LC so that you can get a “feel” for how the LC performs normally.
This will help you to quickly locate problems. For example, often you can smell a leak before you see it. The majority of problems are identified by sight; most of these are included in the preceding section.
Solvent Smell
“Hot” Smell
|
Possible
Cause
|
Prevention/Solution
|
|
Overheating
Module
|
Check for
proper ventilation and adjust it.
Check for temp.
setting and adjust it.
Shut module
off
|
Abnormal Meter Readings
|
Possible
Cause
|
Prevention/Solution
|
|
Pressure
Abnormalities
|
Check for
pressure, and troubleshoot it.
|
|
Column oven
problems
|
Check setting
and adjust it.
|
|
Detector lamp
failing
|
Replace lamp
|
Warning Lamps
|
Possible
Cause
|
Prevention/Solution
|
|
Pressure limit
exceeded
|
Check for
blockage
Check for
limit setting and adjust
|
|
Other warning
Lamps
|
See the
service manual of it.
|
Warning Buzzers
|
Possible
Cause
|
Prevention/Solution
|
|
Solvent leak /
spill
|
Locate and
correct
|
|
Other warning
buzzers
|
See service
manual of it.
|
Squeaks and Squeals
|
Possible
Cause
|
Prevention/Solution
|
|
Bearing
failure
|
See service
Manual
|
|
Poor Lubrication
|
Lubricate as
necessary
|
|
Mechanical
wear
|
See service
Manual
|
