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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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Sample Preparation Details Information

Title Headspace Collection
Description We used two methods and assayed to collect the HS of Arabidopsis seedlings after a 7 day-incubation. To quench enzyme activity in Arabidopsis, we treated the seedlings in the following manner: (i) a set of HS vials wrapped in aluminum foil were incubated at 80 °C for 30 min in a windy oven (WFO-600 ND; Eyela, Tokyo, Japan) and (ii) 0.25-M EDTA-NaOH water solution (pH 7.5) was added into another set of vials to attain a final EDTA concentration of 50 mM. Solid CaCl2 was then immediately added to a final concentration of 5 M. Next, 10-μL of n-alkane standard solutions C8-20 (0.8 mg/l) was added to each vial as an internal standard. The vials were closed with magnetic screw-caps and then sonicated (US-108; NSD, Suwa, Japan) at a frequency of 38-Hz for 5 min. Vials without any treatment were prepared and used as controls.

Nine types of SPME fibers were purchased from Supelco (Supelco, PA, USA). A SPME fiber coated with a 100-μm-thick layer of polydimethylsiloxane (PDMS) metal alloy (100-μm PDMS) was finally chosen for VOC profiling (see Results and Discussion section). SPME fibers were coated with (i) 30-μm- or (ii) 7-μm-thick layers of PDMS-fused silica (FS) fiber/stainless steel (SS), (iii) 75-μm-thick layer of carboxen/polydimethylsiloxane (CAR/PDMS) FS/SS, (iv) 85-μm-thick layer of CAR/PDMS StableFlex (SF) fiber/SS, (v) 65–μm-thick layer of PDMS/divinylbenzene (DVB) SF/SS, (vi) 50/30-μm-thick layer of DVB/CAR on PDMS (DVB/CAR/PDMS) SF/SS, (vii) 85-μm-thick layer of polyacrylate (PA 85) or (viii) carbowax-polyethylene glycol (C-PEG) for HS collection. Before the analysis, all fibers were conditioned at the appropriate conditioning temperature and time as shown in Table 3. Each fiber was exposed to the vial headspace for 20 min at 60 °C with continuous agitation.


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