Why are there signals in the blank or in the zero samples, why are analytes in the zero samples above LOD?
The blank well A1 does not contain any analytes or internal standards. The zero samples only contain internal standards, but no analytes. If there are still peaks the following may have occurred:
- A sample or internal standard mix was accidentally loaded.
- Spillover when loading the extraction solvent onto the kit plate, e.g. the dispensing speed of the repeater was too high.
- Spillover when shaking the plate after loading the extraction solvent.
- Cross-contamination by residues at the bottom of the filter plate when removing the filter plate from the capture plate.
- Cross-contamination when transferring the extracts to make the LC and FIA plates.
- To make sure the contamination comes from the plate and not from the system, inject a blank from an autosampler vial to cross-check.
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How is LOD calculated?
The limit of detection (LOD) for each metabolite within a kit run is calculated from the concentrations in the zero samples based on either the standard deviation (StdDev-based) or median (Median-based). StdDev-based: LOD is calculated as median ...
Why are QC accuracies consistently low or high for all analytes?
If the QC accuracies are consistent for all measured analytes, but differ significantly from the expected values (i.e. all metabolites for any particular QC sample are at 50% accuracy), the following may have occurred: 1. Pipetting issue: More or ...
Why do extracts appear cloudy?
For AbsoluteIDQ® p180, p400 HR and MxP® Quant 500 kits: When adding water to the methanolic sample extracts to make the LC plate, cloudiness can occur due to residues of pyridine and phenyl isothiocyanate from the derivatization solution. If that ...
Why the extracts did not go through the filters or the volumes in the capture plate are low/inconsistent?
Inconsistent or lack of volume in the filter plate may occur for several reasons. Below are some suggestions: Water or another solvent was used for making the extraction solvent instead of Methanol. Perform the following steps: Transfer the extracts ...