Simultaneous column chromatographic extraction and purification of abscisic acid in peanut plants for direct HPLC analysis.
Abscisic acid (ABA), a universal signaling molecule, plays important roles in regulating plant growth, development and stress responses. The low contents and complex components in plants make it difficult to be accurately analyzed. A novel one-step sample preparation method for ABA in plants was developed. Fresh peanut (Arachis hypogaea) plant materials were fixed by oven-drying, microwave drying, boiling or Carnoy's fixative, and loaded onto a mini-preparing column. After washed the impurities, ABA was eluted with a small amount of solvent. ABA in plant materials was completely extracted and purified in 2mL solution and directly analyzed by HPLC, with a 99.3% recovery rate. Multiple samples can be simultaneously prepared. Analyses using this method indicated that the endogenous ABA in oven-dried peanut leaves increased 20.2-fold from 1.01 to 20.37μgg(-1) dry weight within 12h and then decreased in 30% polyethylene glycol 6000 treated plants, and increased 3.34-fold from 0.85 to 2.84μgg(-1) dry weight in 5 days and then decreased in soil drought treated plants. The method combined the column chromatographic extraction and solid-phase separation technologies in one step and can completely extracted plant endogenous ABA in a purified and highly concentrated form for direct HPLC analysis.
Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, and the Research and Development Center for Rare Animals, South China Normal University, Guangzhou 510631 China; Guangzhou Huichuan Medical Technology Company, 211 Jinfu Building, 90 Qifu Road, Baiyun District, Guangzhou 510410, China.
Ya-Wen
YW
Zhang
Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, and the Research and Development Center for Rare Animals, South China Normal University, Guangzhou 510631 China.
Wei-Wei
WW
Fan
Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, and the Research and Development Center for Rare Animals, South China Normal University, Guangzhou 510631 China; Guangzhou Huichuan Medical Technology Company, 211 Jinfu Building, 90 Qifu Road, Baiyun District, Guangzhou 510410, China.
Hui
H
Li
Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, and the Research and Development Center for Rare Animals, South China Normal University, Guangzhou 510631 China.
He
H
Ni
College of Chemistry and Life Sciences, Guangdong University of Petrochemical Technology, 139 Guandu Road 2, Maoming City, Guangdong 525000, China.
Han-Bing
HB
Han
Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, and the Research and Development Center for Rare Animals, South China Normal University, Guangzhou 510631 China. Electronic address: li_haihang@yahoo.com.
Hai-Hang
HH
Li
D000040:Abscisic Acid / Q000032:analysis / Q000302:isolation & purification*; D010367:Arachis / Q000737:chemistry* / Q000502:physiology; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D055864:Droughts; D013312:Stress, Physiological
D000040:Abscisic Acid
Abscisic acid; Extraction; Peanut; Phytohormone; Purification; Sample preparation