HPLC · Calibration procedure
Column oven temperature accuracy
Verifies the column oven holds the set temperature, using a calibrated temperature probe.
PURPOSE
Why this is checked
Retention time and selectivity change with temperature. A drifting oven makes retention times and resolution unreliable.
Frequency
Typically every 6 months, and after major repair or relocation, as defined in your calibration SOP.
Learn how the HPLC worksYOU WILL NEED
Standards & tools
- Calibrated digital thermometer with a thin probe, traceable to national standards
BEFORE YOU START
Prerequisites
- Place the probe in the column compartment at the column position
- Close the oven door and let it stabilise
PROCEDURE
Step by step
- 1Set the oven to a low temperature (e.g. 30 °C) and wait at least 15 minutes after it reaches the set point.
- 2Record the probe reading.
- 3Repeat at a mid and a high temperature used by your methods (e.g. 40 °C and 60 °C).
RECORD
Example observation table
| Set (°C) | Observed (°C) | Difference (°C) |
|---|---|---|
| 30 | 30.4 | +0.4 |
| 40 | 39.8 | −0.2 |
| 60 | 59.3 | −0.7 |
CALCULATION
How to calculate the result
Difference = Observed − Set
- Set 60 °C, observed 59.3 °C
- Difference = −0.7 °C (within ±2 °C)
ACCEPTANCE CRITERIA
Pass if
- Observed temperature within ±2 °C of the set value.
IF IT FAILS
What to do
- Check the door seal and probe position
- Service the oven heater or sensor
- Repeat after repair
MORE
Other HPLC calibrations
In LimsPulse · Calibration Management
LimsPulse schedules this calibration, records the result against limits, and blocks the instrument everywhere if it fails or goes overdue.