Validation | features

Quality assessment

For an easy quality assessment of a kit run, validation statuses are given on metabolite, well/injection, and plate run levels.

• Quality assessment is sample type and analyte classification (T1 or T2) specific.
• "Excluded" metabolites, injections, or wells are not used for quality assessment.

Classification levels

• Metabolite: a calculated concentration for a metabolite is evaluated.
• Injection: all concentrations of an injection are evaluated.
• Well: all injections from one well are evaluated. If only one injection per well was performed, injection level = well level.
• Plate run: all wells of one plate run are evaluated.

Custom QC: "QC" and "Custom QC" samples are grouped under the term "QC".

Validation criteria are only given if not described in section Validation statuses.

Metabolite level

Calibration standard acceptance criteria

• accuracy within kit acceptance criteria, in general 70-130%
• accuracy not within kit acceptance criteria, in general 70-130%

QC sample acceptance criteria

• Accuracy calculated exclusively for metabolites of analyte classification "T1".
• CV calculated exclusively for metabolites of QCs and of analyte classification "T1" or "T2".
  Requirement: at least three QC replicates per plate run and concentration above LOQ.

• accuracy within kit acceptance criteria, in general 70-130%, and CV below 20% (analyte classification T1) or below 30% (analyte classification T2)
• accuracy not within kit acceptance criteria, in general 70-130%
• CV above 20% (analyte classification T1) or above 30% (analyte classification T2)

Well/injection level

Well/injection status is , if

at least 85% metabolites per well/injection with an analyte classification T1 are and

at least 80% metabolites per well/injection with an analyte classification T2 are .

For this evaluation, Valid means for

• T1 metabolites do not have Accuracy ot ouf range or CV out of range,
• T2 metabolites do not have CV out of range.

Plate run level

Plate run validation requirement: at least three QC replicates of one level per plate run.

For plate run validation, QCs are exclusively used.

• OK if at least 67% of all wells per plate run have a well status
• Not OK if less than 67% of all wells per plate run have a well status

Valid well status means QC samples do not have CV out of range or Accuracy out of range, as described in Well/injection level.

Exclude metabolite, well, sample type, or plate run

A metabolite, sample, well, or plate run can be excluded from analysis. This may be required for issues during preparation or data collection, e.g. when a sample was pipetted incorrectly onto one well position or the incorrect retention time (RT) was defined for one metabolite in the corresponding acquisition method.

Exclude a metabolite

1. Select a well, e.g. QC2 on position E2.

   

2. Right click on a metabolite, e.g. Ala.

   

3. Add a note (e.g. "Incorrect RT defined") and click Save.

   

Multiple metabolites can be selected. Click in the field Metabolite. Make a selection, and click Save.

Exclude a well

1. Right click on a well, e.g. QC2 on position E2.

   

2. Add a note (e.g. "Incorrect pipetting") and click Save.

   

• Multiple wells can be selected. Click in the field Well. Make a selection, and click Save.

  

• Samples from the same type (e.g. all QCs) are selected by selecting QCs.

  

Exclude a plate run

1. Above the plate, click Exclude.

   

2. Add a note (e.g. "Kit run failed") and click Save.

   

Add notes

Notes can be added on metabolite, well, or plate run level.

1. Select a metabolite or well and right click, e.g. QC2 on position E2.

   

2. Add a note and click Save.

   

• Plate notes can be added, by right click on .

  

Multiple metabolites or wells can be selected. Click in the field Metabolite or Well, make a selection, and click Save.