Summary of Study ST002136
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 PR001353. The data can be accessed directly via it's Project DOI: 10.21228/M8TQ43 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 | ST002136 |
| Study Title | Targeted Microchip Capillary Electrophoresis-Orbitrap Mass Spectrometry Metabolomics to Monitor Ovarian Cancer Progression (calibration standards) |
| Study Summary | The lack of effective screening strategies for high-grade serous carcinoma (HGSC), a subtype of ovarian cancer (OC) responsible for 80% of OC related deaths, emphasizes the need for new diagnostic markers and a better understanding of disease pathogenesis. Capillary electrophoresis (CE) coupled with high-resolution mass spectrometry (HRMS) offers high selectivity and sensitivity, thereby increasing metabolite coverage and consequently enhancing biomarker discovery. Recent advances in CE-MS include small, chip-based CE systems coupled with nanoelectrospray ionization (nanoESI) to provide rapid, high-resolution analysis of biological specimens. Here, we describe the development of a targeted microchip (µ) CE-HRMS method to analyze 40 target metabolites in serum samples from a triple-mutant (TKO) mouse model of HGSC, with an acquisition time of only 3 min. Extracted ion electropherograms showed sharp, highly resolved peak shapes, even for structural isomers such as leucine and isoleucine. All analytes maintained good linearity with an average R2 of 0.994, while detection limits were in the nM range. Thirty metabolites were detected in mice serum, with recoveries ranging from 78 to 120 %, indicating minimal ionization suppression and good accuracy. We applied the µCE-HRMS method to sequentially-collected serum samples from TKO and TKO-control mice. Time-resolved analysis revealed characteristic temporal trends for amino acids, nucleosides, and amino acids derivatives associated with HGSC progression. Comparison of the µCE-HRMS dataset with non-targeted ultra-high performance liquid chromatography (UHPLC) – MS results revealed identical temporal trends for the 5 metabolites detected on both platforms, indicating the µCE-HRMS method performed satisfactorily in terms of capturing metabolic reprogramming due to HGSC progression, while reducing the total analysis time 3-fold. |
| Institute | Georgia Institute of Technology |
| Last Name | Sah |
| First Name | Samyukta |
| Address | 901 Atlantic Dr NW, Atlanta, GA, 30332, USA |
| ssah9@gatech.edu | |
| Phone | 5746780124 |
| Submit Date | 2022-04-11 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2022-05-02 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN003497 |
|---|---|
| Analysis type | MS |
| Chromatography type | CE |
| Chromatography system | ZipChip (908 Devices) |
| Column | HS chip |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Exactive Plus Orbitrap |
| Ion Mode | POSITIVE |
| Units | Micromolars |
MS:
| MS ID: | MS003257 |
| Analysis ID: | AN003497 |
| Instrument Name: | Thermo Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | All experiments were performed in positive ionization mode in the 50-500 m/z range at a mass resolution setting of 17,500. The capillary temperature was set to 200 °C and the sheath gas flow rate was 2 psi. The automatic gain control (AGC) target value was set to 3E6 and the maximum injection time was 20 ms. Data were acquired using Xcalibur 3.0 (Thermo Scientific) and were imported to Skyline software14 for peak picking and integration. The peak picking procedure used the analyte accurate m/z and migration time. Peak areas obtained from Skyline were exported as spreadsheets for further analysis. Quantitation was performed with the analyte peak areas relative to the peak area of one of the three isotopically labeled internal standards (13C6 arginine, 13C methionine D3 and 13C phenylalanine) chosen based on migration time similarities. |
| Ion Mode: | POSITIVE |
