Six biological replicates of seeds of the different maize lines were used for metabolite profiling in this study. Maize seed grains of each line were ground in liquid N2. The total metabolites were extracted with 70% aqueous methanol from 100 mg of seed powder overnight at 4°C. Following centrifugation at 10,000 g for 10 min, the extracts were absorbed by a CNWBOND Carbon-GCB SPE Cartridge (ANPEL, Shanghai, China) and filtered before UPLC (ultra performance liquid chromatography)-MS/MS analysis.
Carbon gcb spe cartridge
Manufactured by ANPEL
Sourced in China
The Carbon-GCB SPE Cartridge is a solid-phase extraction (SPE) cartridge designed for sample preparation. It contains a combination of carbon and graphitized carbon black (GCB) sorbents, which can be used for the selective retention and extraction of analytes from complex matrices. The core function of this cartridge is to facilitate the isolation, purification, and concentration of target compounds prior to instrumental analysis.
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6 protocols using carbon gcb spe cartridge
1
Metabolite Profiling of Maize Seeds
2
Comprehensive Metabolite Extraction Protocol
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The frozen pulp was crushed using a mixer mill (MM 400; Retsch, Germany) with a zirconia bead for 1.5 min at 30 Hz. Sample powder of 100 mg was weighted and extracted overnight at 4°C with 1.0 ml 70% aqueous methanol, vortexed for three times during the period to increase the extraction efficiency. After be centrifuged at 10,000 g for 10 min, the supernatant was collected, passed through a Carbon‐GCB SPE Cartridge (250 mg, 3 ml, CNWBOND, ANPEL). Before LC‐MS analysis, each sample was filtrated (SCAA‐104, 0.22 μm pore size; ANPEL, http://www.anpel.com.cn/ ).
3
Metabolite Extraction from Freeze-Dried Leaves
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First, the freeze-dried leaves were crushed at 30 Hz for 15 min using a mixer mill (MM 400, Retsch, Haan, Germany) with a zirconia bead. Next, 100 mg of the leaf powder was mixed with 1.0 ml of 70% aqueous methanol and incubated overnight at 4 °C for metabolite extraction. Next, the extracts were centrifuged at 10,000 g for 10 min, absorbed using a Carbon-GCB SPE Cartridge (ANPEL, Shanghai, China), and filtrated using SCAA-104 filter (ANPEL) before liquid chromatography-mass spectrometry analysis.
4
Extraction of Metabolites from Plant Tissue
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The pulp was frozen by liquid nitrogen and crushed using a mixer mill (MM 400; Retsch) with a zirconia bead for 1.5 min at 30 Hz. Sample powder (100 mg) was added to 1.0 ml 70% aqueous methanol and extracted overnight at 4°C. For increasing the extraction efficiency, each sample was vortexed for three times during the period. After a centrifugation at 10,000 g 10 min, the supernatant was collected, filtered using a Carbon‐GCB SPE Cartridge (250 mg, 3 ml, CNWBOND, ANPEL) and each sample was filtrated (SCAA‐104, 0.22 μm pore size; ANPEL, http://www.anpel.com.cn/ ) before LC‐MS analysis.
5
Freeze-dried Leaf Metabolite Extraction
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The frozen leaves were first freeze-dried and then ground to fine powder using a mixer mill (MM 400, Retsch, Germany) with a zirconia bead for 1.5 min at 30 Hz. The powder (100 mg) was treated with 1 ml of 70% aqueous methanol overnight at 4°C. Following centrifugation at 10,000 ×g for 10 min, the extracts in supernatant fraction were absorbed using a CNWBOND Carbon-GCB SPE Cartridge (ANPEL, China) and filtered through a 0.22-μm filter (Millipore, USA) for liquid chromatography-mass spectrometry (LC-MS) analysis [21 (link)].
6
Metabolite Profiling of Maize Seeds
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Six biological replicates of seeds of the 7 studied maize lines were used for metabolite profiling. Maize seed grains of each line were ground in liquid N 2 . The total metabolites were extracted from 100 mg of seed powder with 70% aqueous methanol overnight at 4 °C. Following centrifugation at 10,000g for 10 min, the extracts were absorbed by a CNWBOND Carbon-GCB SPE Cartridge (ANPEL, Shanghai, China) and filtered before UPLC (ultra-performance liquid chromatography)-MS/MS (mass spectrum) analysis.
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