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Sample Set Information

ID TSE1236
Title Development of a Direct Headspace Collection Method from Arabidopsis Seedlings Using HS-SPME-GC-TOF-MS Analysis.
Description Plants produce various volatile organic compounds (VOCs), which are thought to be a crucial factor in their interactions with harmful insects, plants and animals. Composition of VOCs may differ when plants are grown under different nutrient conditions, i.e., macronutrient-deficient conditions. However, in plants, relationships between macronutrient assimilation and VOC composition remain unclear. In order to identify the kinds of VOCs that can be emitted when plants are grown under various environmental conditions, we established a conventional method for VOC profiling in Arabidopsis thaliana (Arabidopsis) involving headspace-solid-phase microextraction-gas chromatography-time-of-flight-mass spectrometry (HS-SPME-GC-TOF-MS). We grew Arabidopsis seedlings in an HS vial to directly perform HS analysis. To maximize the analytical performance of VOCs, we optimized the extraction method and the analytical conditions of HP-SPME-GC-TOF-MS. Using the optimized method, we conducted VOC profiling of Arabidopsis seedlings, which were grown under two different nutrition conditions, nutrition-rich and nutrition-deficient conditions. The VOC profiles clearly showed a distinct pattern with respect to each condition. This study suggests that HS-SPME-GC-TOF-MS analysis has immense potential to detect changes in the levels of VOCs in not only Arabidopsis, but other plants grown under various environmental conditions.
Authors Kusano M, Iizuka Y, Kobayashi M, Fukushima A, Saito K.
Reference Metabolites. 2013 Apr 9;3(2):223-42. doi: 10.3390/metabo3020223.

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Analytical Method Details Information

Instrument GC:Agilent 6890N MS:LECO Pegasus 4D MS system
Instrument Type
Ionization EI
Ion Mode Positive
Description After HS collection, the volatiles were thermally desorbed in splitless mode on a CTC CombiPAL autosampler (CTC analytics, Zwingen, Switzerland) connected to an Agilent 6890N gas chromatograph (Agilent Technologies, Wilmington, USA) for 0.1 min at the appropriate inlet temperature, as shown below. Each fiber was baked for 5 min by applying the appropriate conditioning temperature (Table 3).

The capillary column used for the analysis was a 30-m×0.25-mm inner diameter fused-silica capillary column with a chemically bound 0.25-μL film Rxi-5 Sil MS stationary phase (RESTEK, Bellefonte, USA) with a tandem connection to a fused silica tube (1 m, 0.15 mm). A mass spectrometer column change interface (ms NoVent-J; SGE, Yokohama, Japan) was used to prevent air and water from entering the MS during column change over. Helium was used as the carrier gas at a constant flow rate of 1.0 ml/min. The temperature program started with a 2 min isothermal step at 50 °C, followed by temperature ramping at 15 °C to a final temperature of 260 °C, which was then maintained for 2 min. The transfer line to the mass spectrometer was set to 250 °C. The TOF mass spectrometer was a Pegasus 4D MS system (Leco, MI, USA) with an EI source set to 200 °C. The acceleration voltage was turned on after a solvent delay of 200 s. Mass spectra were monitored with an acquisition rate of 30 spectra/s and over a mass-to-charge ratio range of m/z = 30–550.


Table 3. Conditioning temperature and time for the SPME fibers used in the study.
SPME fiber Conditioning temperature(°C) Conditioning time(h) Inlet temperature(°C)
100-μm PDMS 250 0.5 220
30-μm PDMS 250 0.5 220
7-μm PDMS 320 0.5 220
85-μm polyacrylate 280 1 220
60-μm PEG 240 0.5 220
75-μm CAR/PDMS 300 1 280
85-μm CAR/PDMS 300 1 280
65-μm PDMS/DVB 250 0.5 250
50/30-μm DVB/CAR/PDMS 270 0.5 250
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