calibration of HPLC

¿What is the calibration of HPLC?

The calibration of HPLC

In the calibration of hplc we perform the flow of all line (1 line or 4 lines) and then run the GPV test.

After that we perform the following test

  • Injector linerity
  • Injector precision
  • Wave length calibration
  • Detector linerity
  • Temperature checking for both sample compartment and CTO
  • Leak sensor check
  • Autosampler Vail test.

In the metrology sense where calibration means comparison against a known reference standard, you only actually calibrate wavelength accuracies, temperatures, and flow. In the lab setting you also do several performance verifications depending on your system configuration and methodologies.

They could include:
Detector linearity
Injector precision
Injector linearity
Gradient accuracy

Calibration basically suggests that decisive the link between system response and analyte quantity. Closely tied to this is the concept of "system suitability". In a regulated environment (GLP or cGMP), any validated method should have a set of system suitability criteria. These will typically include things like retention time(s), plate numbers, RSDs, etc. These obviously require the use of a column.

From the context, "qualification" (specifically, "performance qualification" or "PQ"), which is verifying that the instrument if functioning properly within its specifications. In a regulated environment, this is typically done at least annually. Complete PQ on associate degree HPLC system involves confirmatory things like

- intake line integrity
- check valve/seal integrity
- flow accuracy
- proportioning accuracy/dwell volume (gradient systems)
- reproducibility & linearity
- noise & drift
- wavelength accuracy
Some of these tests involve the pump alone; some (proportioning or dwell) require a restrictor between injector and detector; some require a column (injector tests) and for some (detector) either way will work.


A. Wikipedia:

B. Knauer:

C. Kromasil:

D. Shimadzu:

E. ChemistryView:


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