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Then, 50 l of reaction mixture was sequentially treated with 50 l of 50 mM 3-NPH (Sigma) in methanol and H 2 O (70:30, v/v), 50 l of 50 mM EDC (Oakwood Chemical) in methanol and H 2 O (70:30, v/v) and 50 l of 7% v/v pyridine (Acros Organics) in methanol and H 2 O (70:30, v/v) and mixed thoroughly