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Following four to five washes with water, the material was freeze dried.
That is, the entire material was freeze dried after AHP treatment and used as the substrate for enzymatic hydrolysis.
For the determination of GSH, cysteine, cysteine glycine and γ-glutamate-cysteine (reduced, oxidized and total), the plant material was freeze dried before grinding into a fine powder with a pestle and mortar using liquid nitrogen.
For this the leaf material was freeze dried, and 10 mg samples were subjected to elemental analysis using the Vario EL cube (Elementar Analysensysteme GmbH, Germany) in the CN operation mode.
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Chimney material was frozen at −80°C immediately after collection and remained frozen until onshore analysis.
The material was freeze-dried in two cycles using a freeze-dryer Epsilon 2-10D (Martin Christ GmbH, Osterode, Germany).
Briefly, leaf material was freeze-dried for 48 h and homogenized with a mill (Retsch MM 200) to fine powder.
Urine supernatant (1 mL) was stored at -20 °C for NMR measurement, and the remaining material was freeze-dried to provide urine powder for further analysis.
Water-soluble and chloroform-butanol layer were separated by centrifugation, an equal amount of ethanol was added to the water-soluble layer (this procedure was repeated twice), and the ethanol-precipitated material was freeze-dried and stored at -80°C until use [ 19].
Plant material was frozen in liquid nitrogen.
The material was frozen and stored at -80°C.
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