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They were then mixed with 1 mL of a 40:40:20 methanol: acetonitrile: water solution precooled to 20 C, followed by vortexing for 5 min, incubation at 20 C for 10 min, and centrifugation at 4 C and 12,000 g for 15 min

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Very Low), and T2DM (Class IV

its a critical variable that can define the parameters and outcomes of an entire study