Two-dimensional chromatography based on on-line HPLC-DPPH bioactivity-guided assay for the preparative isolation of analogue antioxidant compound from Arenaria kansuensis.
Traditional Tibetan medicine is important for discovery of drug precursors. However, information about the chemical composition of traditional Tibetan medicine is very limited due to the lack of appropriate chromatographic purification methods. In the present work, A. kansuensis was taken as an example and a novel two-dimensional reversed-phase/hydrophilic interaction liquid chromatography(HILIC) method based on on-line HPLC-DPPH bioactivity-guided assay was developed for the purification of analogue antioxidant compounds with high purity from the extract of A. kansuensis. Based on the separation results of many different chromatographic stationary phases, the first-dimensional (1D) preparation was carried on a RP-C18HCE prep column, and 2 antioxidant fractions were obtained from the 800mg crude sample with a recovery of 56.7%. A HILIC-XAmide prep column was selected for the second-dimensional (2D) preparation. Finally, a novel antioxidant β-carboline Alkaloids (Glusodichotomine AK) and 4 known compounds (Tricin, Homoeriodictyol, Luteolin, Glucodichotomine B) were purified from A. kansuensis. The purity of the compounds isolated from the crude extract was >98%, which indicated that the method built in this work was efficient to manufacture single analogue antioxidant compounds of high purity from the extract of A. kansuensis. Additionally, this method showed great potential in the preparation of analogue structure antioxidant compounds and can serve as a good example for the purification of analogue structure antioxidant carboline alkaloids and flavonoids from other plant materials.
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Yulei
Y
Cui
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Na
N
Shen
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Xiang
X
Yuan
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China.
Jun
J
Dang
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China.
Yun
Y
Shao
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China.
Lijuan
L
Mei
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China. Electronic address: m17701159965@163.com.
Yanduo
Y
Tao
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China.
Qilan
Q
Wang
Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research, Chinese Academy of Sciences, Xining 810008, Qinghai, PR China; Key Laboratory of Tibetan Medicine Research of Qinghai Province, Xining 810008, Qinghai, PR China. Electronic address: XBGYSWYJS@163.com.
Zenggen
Z
Liu
D000470:Alkaloids / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D000975:Antioxidants / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D031243:Arenaria Plant / Q000737:chemistry*; D001713:Biphenyl Compounds / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D010851:Picrates / Q000737:chemistry; D010936:Plant Extracts / Q000032:analysis* / Q000737:chemistry
D000470:Alkaloids; D000975:Antioxidants; D001713:Biphenyl Compounds; D010851:Picrates; D010936:Plant Extracts; C004931:1,1-diphenyl-2-picrylhydrazyl
Antioxidant; Arenaria kansuensis; HPLC-DPPH-DAD; Two-dimensional separation