High-Performance Liquid Chromatographic Quantification of the Plant Hormone Abscisic Acid at ppb Levels in Plant Samples after a Single Immunoaffinity Column Cleanup.
Jiaqi ZhangMan ZhangMian WangYixuan WuYang ShiYujie ChenRui FengXiaoling YangXiaojiao ChenBaomin WangPublished in: Journal of agricultural and food chemistry (2024)
High-performance liquid chromatography with ultraviolet detection (HPLC-UV) is a common analysis technique due to its high versatility and simple operation. In the present study, HPLC-UV detection was integrated with immunoaffinity cleanup (IAC) of the sample extracts. The matrix effect was greatly reduced, and the limit of detection was as low as 1 ng/g of free abscisic acid (ABA) in fresh plant tissues. A monoclonal antibody 3F1 (mAb 3F1) was developed to specifically recognize free ABA but not ABA analogues. The mAb 3F1-immobilized immunoaffinity column exhibited a capacity of 850 ng/mL and an elution efficiency of 88.8-105% for standards. The extraction recoveries of the column for ABA ranged from 80.4 to 108.9%. ABA content was detected in various plant samples with IAC-HPLC-UV. The results were verified with ultraperformance liquid chromatography-electrospray tandem mass spectrometry. IAC-HPLC-UV can be a sensitive and cost-efficient method for plant hormone analysis.
Keyphrases
- liquid chromatography
- tandem mass spectrometry
- high performance liquid chromatography
- solid phase extraction
- simultaneous determination
- ultra high performance liquid chromatography
- mass spectrometry
- liquid chromatography tandem mass spectrometry
- high resolution mass spectrometry
- monoclonal antibody
- gas chromatography
- transcription factor
- arabidopsis thaliana
- ms ms
- cell wall
- real time pcr
- data analysis
- capillary electrophoresis
- molecular dynamics simulations