How to Use FIx Analysis for Peak Normalization and How to Align Peaks
Peak Normalization
Peak normalization refers to the process of scaling the peak areas or heights across different samples to a common reference, enabling meaningful comparison. This procedure helps minimize variability arising from differences in sample concentration or instrumental performance. Common normalization methods include:
1. Total Area Normalization
The total peak area of each sample is scaled to a fixed reference value (e.g., 100 or 1), and each individual peak is proportionally adjusted to reflect this scaling.
2. Maximum Peak Normalization
The most intense peak in each sample is identified and set to a predefined value (e.g., 100 or 1). All other peak areas are then adjusted proportionally based on this reference.
3. Internal Standard Normalization
An internal standard compound with a known concentration is added to each sample. The peak areas of other analytes are then normalized relative to the internal standard’s peak area.
In the FIx Analysis software, normalization options are available within the data processing module. Users can select the preferred normalization method and enter the required parameters as instructed by the software interface.
Peak Alignment
Peak alignment refers to adjusting the retention time or mass of corresponding peaks across samples to achieve consistency and enable direct comparison. The most commonly employed approaches include:
1. Retention Time Correction
Software algorithms compare retention times across different samples and adjust them according to predefined criteria or statistical models to align corresponding peaks.
2. Alignment Algorithms
Techniques such as Dynamic Time Warping (DTW) or mass spectral matching are employed to achieve precise alignment based on peak shape and other features.
To perform peak alignment in FIx Analysis, navigate to the alignment module, select an appropriate algorithm, and configure relevant parameters (e.g., alignment window size, alignment thresholds, etc.). The software typically provides a preview function to inspect the alignment results before applying them to the full dataset.
These two processes are critical to ensuring the quality and comparability of analytical data. For detailed operational guidance, refer to the FIx Analysis user manual or consult technical support, as procedural details may vary between software versions.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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