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It was heated at 117 C for 10 min (HMC Autoclave HG80, Germany) to precipitate the proteins contained therein

The mobile phase consisted of solvent A (0.1% formic acid-acetonitrile/water (3:2 v/v)) and solvent B (0.1% formic acid-isopropanol/acetonitrile (9:1 v/v)) with a gradient elution (01.5 min, 6868% A

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