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Total of 20 L 10% O-pthaldialdehyde (OPA) in methanol was added to each assay mixture, vials were capped, the resulting reaction mixture incubated at 25 C for 5 min in the dark and analysed using high performance liquid chromatography (HPLC) (Merck KGaA, Darmstadt, Germany)

Use a different area the next day to reduce repeated exposure to the same spot

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