Metabolites were extracted from the deep frozen cells by a pre-mixture of 300 µl methanol (final concentration ~80 %), 30 µl nonadeconoic acid methylester (2 mg ml-1 chloroform) and 30 µl of ribitol (0.2 mg ml-1), and D4-2,3,3,3-alanine (1 mg ml-1, standards in methanol). Each sample was mixed thoroughly at least 1 min for complete suspension, agitated 15 min at 70°C, brought to room temperature mixed with 200 µl chloroform and agitated again 5 min at 37°C.
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