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Monitoring

Monitoring provides additional tools to monitor the LC-MS and kit performance.

Monitoring

Analytical assessment​

Summarizes results from Validation.

Analytical assessment

Evaluation is based on currently valid EMA Guideline for bioanalytical method validation and available for QCs and calibration standards.

QC or calibration standards

  • Metabolites are classified as "valid" or "invalid", based on kit validation criteria applied in Validation.
  • For analyte evaluation of QCs these criteria are applied: 66% of replicates of one QC level and minimum 50% of all QC levels must be valid.

Example | Replicates of one QC level

At least 66% are valid. The resulting Analyte evaluation is valid. Analyte classification valid

Less than 66% are valid. The resulting Analyte evaluation is invalid. Analyte classification invalid

Monitoring​

Monitors the LC-MS instrument and kit performances over a specified time period.

Monitoring

  1. Select a value which should be displayed, e.g. "Analyte intensity [cps]".
    Select a value
  2. Select a sample type, QC2 is selected by default.
    Select a sample
  3. Define a time period from which data is loaded.
    Define time period
  4. Select an OP from which data is loaded, e.g. MXP500L-0-5813 for Quant 500 kit, LC part, SCIEX 5500+.
    Select OP
  5. Select a metabolite, e.g. Ala.
    Select metabolite

A plot is shown. To monitor for example the instrument sensitivity by time, use the value Analyte intensity [cps] of QC2 samples. If the intensity decreases significantly, a calibration or cleaning of the instrument may be required.

Plot

Plots​

Use plots to evaluate the accuracies and CVs of a plate run.

Plots

  1. Select a plate run, e.g. LC part of a Quant 500 kit.
    Select plate run
  2. Select a sample type, e.g. QC2.
    Select a sample

Accuracies and CVs are shown as graphics and violin plots. Examples are shown below.

Graphic accuracy Plot CV

Detectability​

Check the metabolite detectability and analytical performance.

Detectability

The options < LOD and Accuracy are available.

Metabolite detectability

  1. Select a plate run, e.g. LC part of a Quant 500 kit.
    Select plate run
  2. Select < LOD.
    Select LOD
  3. Select a sample type, e.g. QC2.
    Select a sample
  4. All Metabolites analyzed with a method of the selected plate run, e.g. 105, and the Detected metabolites, e.g. 102 of the selected sample(s) are shown.
    Detectability results

Metabolites with concentrations "< LOD" are highlighted in red.

Metabolite &lt;LOD

Accuracy check

  1. Select a plate run, e.g. LC part of a Quant 500 kit.
    Select plate run
  2. Select Accuracy.
    Select Accuracy
  3. Select a sample type, e.g. QC2.
    Select a sample
  4. Define an acceptance threshold, e.g. 20%
    Acceptance threshold
  5. All Metabolites with the analyte classification "quantitative" (T1) of the selected plate run, e.g. 42, and those meeting the acceptance threshold, Detected metabolites, e.g. 40, of the selected sample(s) are shown.
    Accuracy results

Metabolites with accuracies outside the defined acceptance threshold are highlighted in red.

Metabolite &lt;LOD