Accelerating Research in Life Sciences PREMIER Biosoft Accelerating Research in Life Sciences

A High Throughput Lipid Characterization Tool using SONAR Data

Major Features Database containing 40,298 lipid structures and 15,09,305 structure-specific in-silico MS/MS characteristic ions. Load raw data from native files (.raw) for Triple Quadrupole, and SONAR workflows. Data processing – peak detection, peak picking – for LC-, SFC- SONAR workflows.

Major Features Visualize raw data, and processed data – TIC, spectra, XIC of detected compound in retention time scale, XIC of compound in quadrupole positions (bin) scale, XICs overlay of precursor ion m/z, and its fragment ions in quadrupole positions (bin) scale. High throughput lipid structural identification using exact mass, and product ions from SONAR workflows. Automated annotation of MS/MS spectra with IDed lipids, and their fragments.

Create a New Project 1. File > New > Project

Create a New Project 2. Enter a project name 3. Click Create

Import Data from Native as well as Standard Data Files 2. Choose the type of file 1. File > Open > Peaklist Data > Waters Data File > Waters (DIA) > SONAR (.raw)

Use the Shortcut to Import Waters Native File as Shown Below 1. Select Waters native file shortcut icon

Use the Shortcut to Import Waters Native File as Shown Below 1. Select Waters native file shortcut icon 2. Select the file(s) 3. Click Open

Specify Range 1. Select Profile Data 2. Press OK

Retrieving Selected Scan Number Information

The Selected Files/Scans can be Viewed from the Project Management Panel Along with Chromatogram and Mass Spectrum Loaded Data

The Selected Files/Scans can be Viewed from the Project Management Panel Along with Chromatogram and Mass Spectrum Tabular View Chromatogram view Loaded data selected Select row Search Results/ Mass Annotation & Lipid information are not applicable for DIA data Spectrum View

Chromatogram and Mass Spectrum Zoom Options 1. Pinned Chromatogram view

Chromatogram and Mass Spectrum Zoom Options 2. Select settings for zooming 3. Click OK

Chromatogram and Mass Spectrum Zoom Options Selecting Chromatogram area for zoom

Chromatogram and Mass Spectrum Zoom Options Pinned Spectrum View

Chromatogram and Mass Spectrum Zoom Options 2. Select settings for zooming 3. Click OK

Chromatogram and Mass Spectrum Zoom Options Selecting Spectrum area for zoom

Peak Detection and Picking 1. Analyze > DIA Peak Detection > RT + Quad m/z 2. Select Profiles 3. Select Template (UPLC SONAR Lipidomic) 4. Select Settings

Peak Detection and Picking 1. RT Peaks

Peak Detection and Picking 2. Precursor Ions

Peak Detection and Picking 3. Product Ions 4. Press OK

Peak Detection and Picking 5. Select OK

Peak Detection and Picking

CID's are Detected and Displayed under LC-MS and MS/MS Data Node

High Throughput Lipid Search 1. Go to Analyze > LC-MS Pre-processing > High Throughput Lipid Search 3. Select MS/MS Data option to perform HTP search only for MS/MS data 2. Specify Range 4. Click OK

Specify HTP Search Parameters 1. Select profiles 2. Specify Search Parameters 5. Click Search 4. Click OK 3. Specify Filter Lipids

High Throughput Lipid Search 5. Click OK

Check HTP Status and Load Results

A high throughput search ID number is generated HTP ID

Check HTP Status and Load Results Click this button to check HTP search status

Check HTP Status and Load Results HTP search status-Pending

Check HTP Status and Load Results HTP search status-Completed

Check HTP Status and Load Results Load HTP Results

Check HTP Status and Load Results

Search results are displayed as shown below Search Results are displayed here

Mass Spectrum Annotation Sorting by m/z value

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Mass Spectrum Annotation

Generate High Throughput Report 2. Select Profiles 1. File > Generate High Throughput Report 3. Click OK 4. Specify MS/MS Search Parameters 5. Specify the sorting options for organizing results in report 6. Specify the lipid information to be exported in report 7. Select output file type 8. Click OK

Results Exported into MS Excel File