Summary of Study ST002543
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 PR001638. The data can be accessed directly via it's Project DOI: 10.21228/M8ZX4D This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST002543 |
| Study Title | GC/MS analysis of hypoxic volatile metabolic markers in the MDA-MB-231 breast cancer cell line |
| Study Summary | Hypoxia in disease describes persistent low oxygen conditions, observed in a range of pathologies, including cancer. In the discovery of biomarkers in biological models, pathophysiological traits present a source of translatable metabolic products for the diagnosis of disease in humans. Part of the metabolome is represented by its volatile, gaseous fraction; the volatilome. Human volatile profiles, such as those found in breath, are able to diagnose disease, however accurate volatile biomarker discovery is required to target reliable biomarkers to develop new diagnostic tools. Using custom chambers to control oxygen levels and facilitate headspace sampling, the MDA-MB-231 breast cancer cell line was exposed to hypoxia (1% oxygen) for 24 hours. The maintenance of hypoxic conditions in the system was successfully validated over this time period. Targeted and untargeted gas chromatography mass spectrometry approaches revealed four significantly altered volatile organic compounds when compared to control cells. Three compounds were actively consumed by cells: methyl chloride, acetone and n-Hexane. Cells under hypoxia also produced significant amounts of styrene. This work presents a novel methodology for identification of volatile metabolisms under controlled gas conditions with novel observations of volatile metabolisms by breast cancer cells. |
| Institute | University of York |
| Last Name | Issitt |
| First Name | Theo |
| Address | Biology Dept. University of York, Personal |
| ti538@york.ac.uk | |
| Phone | 07398244497 |
| Submit Date | 2023-03-31 |
| Num Groups | 4 |
| Publications | T. Issitt et al., Volatile compounds in human breath: critical review and meta-analysis Journal of Breath Research, Volume 16, Number 2 (2022) https://iopscience.iop.org/article/10.1088/1752-7163/ac5230#jbrac5230s2 |
| Analysis Type Detail | GC-MS |
| Release Date | 2023-04-21 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN004190 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | HP GCD 1800B |
| Column | Agilent PoraBOND Q (25m x 0.32mm x 0.5um) |
| MS Type | EI |
| MS instrument type | Single quadrupole |
| MS instrument name | Agilent/HP 5972 MSD |
| Ion Mode | POSITIVE |
| Units | pg/hr/ug and g/hr for media |
Chromatography:
| Chromatography ID: | CH003104 |
| Chromatography Summary: | Collected canister samples were transferred to a liquid nitrogen trap through pressure differential. Pressure change between beginning and end of “injection” was measured, allowing calculation of the moles of canister collected air injected. Sample in the trap was then transferred, via heated helium flow, to an Aglient/HP 5972 MSD system (Santa Clara, CA, USA) equipped with a PoraBond Q column (25 m x 0.32 mm x 0.5 μm film thickness) (Restek©, Bellefonte, PN, USA). Targeted samples were analyzed in selected ion monitoring (SIM) mode, and untargeted samples in full scan (SCAN) mode with the mass range of 45-200 amu. The mass spectrometer was operated in electron impact ionization mode with 70 eV ionization energy, and transfer line, ion source, and quadrupole temperatures of 250, 280 and 280, respectively. For details on SIM and significantly altered, identified SCAN compounds, see Table 1. All samples were analysed within 6 days of collection. The oven program for both SIM and SCAN analyses were identical and are as follows: 35 ˚C for 2 min, 10 ˚C/min to 155 ˚C, 1 ˚C/min to 131 ˚C and 25 ˚C/min to 250 with a 5 min 30 second hold. |
| Instrument Name: | HP GCD 1800B |
| Column Name: | Agilent PoraBOND Q (25m x 0.32mm x 0.5um) |
| Column Temperature: | 250 |
| Flow Gradient: | NA |
| Flow Rate: | 10ml/min |
| Solvent A: | NA |
| Solvent B: | NA |
| Chromatography Type: | GC |
