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Impacts of citric acid (CA) treatment under non-acidic conditions (pH 7.0, 8.0 and 9. on whey protein isolate (WPI) were examined in this study. Size exclusion chromatography and SDS-PAGE indicated that molecular size and weight of WPI-CA became larger at pH 7.0, 8.0 and 9.0 with CA ranged from 0 to 15 mg/mL, but the protein aggregates disappeared after β-mercaptoethanol was added. The free SH groups of WPI-CA gradually decreased. This could be deduced that CA could promote disulfide bond formation of WPI at the non-acidic pH values.