An off‐line DPPH‐GC‐MS coupling countercurrent chromatography method for screening, identification, and separation of antioxidant compounds in essential oil

Abstract

Essential oils are an important source of natural antioxidants and multiple methods have been established for evaluation of their overall antioxidant activity, however, the antioxidant activities of their compounds are less investigated. In the present study, the hyphenation of 2,2′‐ diphenyl‐1‐picrylhydrazyl (DPPH)‐gas chromatography (GC)‐mass spectrometry (MS) offline and high‐speed countercurrent chromatography (HSCCC) is established for efficient screening, identification, and isolation of antioxidants from essential oils and applied to the essential oil of Curcuma wenyujin Y.H. Chen et C. Ling. Five compounds are preliminarily screened as antioxidants using DPPH‐GC according to the reduction of GC peak areas of each compound after reaction with DPPH and then identified as eucalyptol (7.66%), camphor (2.34%), δ‐elemene (1.15%), β‐elemene (7.10%), and curzerene (15.77%) using GC‐MS. Moreover, these five compounds are isolated by HSCCC using two solvent systems, n‐hexane‐acetonitrile‐ethanol (5:3:2, v/v) and n‐hexane-acetonitrile‐acetone (4:3:1, v/v), and subjected to DPPH scavenging assay. Camphor, δ‐elemene, and β‐elemene show weak DPPH scavenging activity, while curzerene and eucalyptol show moderate DPPH scavenging activity. Notably, a significant synergistic effect on DPPH scavenging is found between curzerene and eucalyptol. The result demonstrated that off‐line DPPH‐GC‐MS coupling CCC is an efficient method for screening, identification, and separation of antioxidant compounds in essential oil.