Summary of Study ST002854
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001787. The data can be accessed directly via it's Project DOI: 10.21228/M8QM78 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST002854 |
| Study Title | HILIC-IM-MS for Simultaneous Lipid and Metabolite Profiling of Microorganisms |
| Study Summary | Progress in the ion mobility mass spectrometry (IM-MS) field has significantly increased our ability to make small molecule and lipid identifications, making it an attractive approach for untargeted multi-omics experiments. The dimensionality of collision cross section (CCS) coupled with tandem mass spectrometry (MS/MS) for feature annotation has become a useful tool for high confidence structural elucidation in complex mixtures in the absence of authentic standards. A comprehensive method for feature identification of small organisms has remained limited to exploring genetic markers and protein signatures, however these methods for identification only scratch the surface of effective methods for bacterial classification. Multi-omic methods that include the metabolome and lipidome have grown in popularity due to the increased capacity for organism specific information. We have achieved species-level identification of Enterococcus faecium, Staphylococcus aureus, Acinetobacter baumannii, and Pseudomonas aeruginosa using a modern single-phase extraction method with hydrophilic interaction liquid chromatography (HILIC) coupled to traveling wave ion mobility mass spectrometry (TWIMS). To test the robustness of this optimized workflow, we included internal standards as a metric for efficiency of the extraction, and well known calibrants for validation for our CCS calibration method. We observed significant differences in metabolite profiles at the strain level using multi-variate statistics, primarily including quorum sensing metabolites in Gram-negative strains, and energy production metabolites in the Gram-positive strains. Lipid profiles showed staggering differences in acyl tail compositions that effectively categorized the microbes, including several classes of phospholipids and glycolipids. We have demonstrated a powerful workflow using multi-dimensional techniques for bacterial speciation in a single injection. |
| Institute | University of Georgia |
| Last Name | Carpenter |
| First Name | Jana |
| Address | 302 E Campus Rd., Athens, Georgia, 30602, USA |
| kelly.hines@uga.edu | |
| Phone | 706-542-1966 |
| Submit Date | 2023-09-07 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-09-27 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN004675 | AN004676 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Waters Acquity | Waters Acquity |
| Column | Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um) | Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Waters Synapt G2 XS QTOF | Waters Synapt G2 XS QTOF |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | internsity | intensity |
MS:
| MS ID: | MS004422 |
| Analysis ID: | AN004675 |
| Instrument Name: | Waters Synapt G2 XS QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | The UPLC was connected to the electrospray ionization source of the traveling wave ion mobility-mass spectrometer (Waters Synapt XS) and samples were injected at 5 uL. Prior to acquisition of sample data, data was acquired for a mixture of CCS calibrants using direct infusion. Randomized sample queues were analyzed in both positive and negative ionization modes. A pooled mixture of all samples was used as a quality control (QC). Data was collected across the entire 17 min chromatographic method using data-independent MS/MS acquisition. Leucine enkephalin was monitored for post-acquisition lockmass correction. Capillary +3 kV; Sampling Cone 30 V; Sampling Cone 25 V; Source Offset 40 V; Source Temp 150 ºC; Desolvation Temp 400 ºC; Cone gas flow 50 L/h; Desolvation gas flow 650 L/h; Nebulizer gas flow 7 Bar. Mass Range 50-1200 m/z. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004423 |
| Analysis ID: | AN004676 |
| Instrument Name: | Waters Synapt G2 XS QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | The UPLC was connected to the electrospray ionization source of the traveling wave ion mobility-mass spectrometer (Waters Synapt XS) and samples were injected at 5 uL. Prior to acquisition of sample data, data was acquired for a mixture of CCS calibrants using direct infusion. Randomized sample queues were analyzed in both positive and negative ionization modes. A pooled mixture of all samples was used as a quality control (QC). Data was collected across the entire 17 min chromatographic method using data-independent MS/MS acquisition. Leucine enkephalin was monitored for post-acquisition lockmass correction. Capillary -2 kV; Sampling Cone 30 V; Sampling Cone 25 V; Source Offset 40 V; Source Temp 150 ºC; Desolvation Temp 400 ºC; Cone gas flow 50 L/h; Desolvation gas flow 650 L/h; Nebulizer gas flow 7 Bar. Mass Range 50-1200 m/z. |
| Ion Mode: | NEGATIVE |
