Fresh colonies from a plate of Columbia agar with 5% sheep's blood were collected after 48 hours' incubation at 37°C for fatty acid analysis. Cellular fatty acid analysis was performed by gas chromatography/mass spectrometry (GC/MS). Two samples were prepared with approximately 100 mg of bacterial biomass each collected from a culture plate. Cellular fatty acid methyl esters were prepared as described by Sasser [19] . GC/MS analyses were carried out as previously described [20] (link). Briefly, fatty acid methyl esters were separated using an Elite 5-MS column and monitored by a Clarus 500 gas chromatograph equipped with a SQ8S MS detector (PerkinElmer, Courtaboeuf, France). Fatty acid methyl esters were identified by using the spectral database search using MS Search 2.0 operated with the Standard Reference Database 1A (National Institute of Standards and Technology, Gaithersburg, MD, USA) and the FAMEs mass spectral database (Wiley, Chichester, UK).
Sq8s ms detector
Manufactured by PerkinElmer
Sourced in France
The SQ8S MS detector is a single quadrupole mass spectrometer designed for routine quantitative and qualitative analysis. It features an enhanced ion optics design for improved sensitivity and robustness.
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Lab products found in correlation
2 protocols using sq8s ms detector
1
Bacterial Fatty Acid Profiling by GC/MS
2
Volatile Organic Compound Profiling
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To measure the temporal production of VOCs in the bottles, different incubation times were tested for each culture condition: 48, 72 and 96 hours at 37 °C. After selecting the most suitable incubation time of 72 hours, VOCs were measured in triplicate for each condition. VOCs were collected for 10 minutes using a PDMS/DVB 65 μm SPME fiber (Supelco, Sigma-Aldrich, Saint-Quentin Fallavier, France) inserted in the headspace of each culture bottle. Gas and liquid chromatography-mass spectrometry (GC/MS and LC/MS) analyses were carried out with a Clarus 500 gas chromatograph equipped with a SQ8S MS detector (Perkin Elmer, Courtaboeuf, France) and with a liquid chromatograph. All data were collected and processed using Turbomass 6.1 (Perkin Elmer). A spectral database search was performed using MS Search 2.0 operated with the Standard Reference Database 1 A (NIST, Gaithersburg, USA). Chemical identifications were validated with both reverse and forward scores above 800. The measurement of a blank sample was carried out in parallel, consisting of a noninoculated blood culture bottle containing the same culture medium as the inoculated bottles. Consequently, the results only considered VOCs measured in culture samples after subtracting the corresponding blank values.
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