单次免疫亲和柱净化后，采用高效液相色谱法定量分析植物样品中十亿分之一水平的植物激素脱落酸。
High-Performance Liquid Chromatographic Quantification of the Plant Hormone Abscisic Acid at ppb Levels in Plant Samples after a Single Immunoaffinity Column Cleanup.
作者信息
Zhang Jiaqi, Zhang Man, Wang Mian, Wu Yixuan, Shi Yang, Chen Yujie, Feng Rui, Yang Xiaoling, Chen Xiaojiao, Wang Baomin
机构信息
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
College of Food and Bioengineering, Xihua University, Chengdu 610039, China.
出版信息
J Agric Food Chem. 2024 May 22;72(20):11794-11803. doi: 10.1021/acs.jafc.4c01680. Epub 2024 May 13.
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.
高效液相色谱-紫外检测法（HPLC-UV）因其通用性强且操作简单，是一种常用的分析技术。在本研究中，HPLC-UV检测与样品提取物的免疫亲和净化（IAC）相结合。基质效应大大降低，新鲜植物组织中游离脱落酸（ABA）的检测限低至1 ng/g。开发了一种单克隆抗体3F1（mAb 3F1），它能特异性识别游离ABA，但不能识别ABA类似物。固定有mAb 3F1的免疫亲和柱对标准品的容量为850 ng/mL，洗脱效率为88.8-105%。该柱对ABA的提取回收率在80.4%至108.9%之间。采用IAC-HPLC-UV检测了各种植物样品中的ABA含量。结果通过超高效液相色谱-电喷雾串联质谱法进行了验证。IAC-HPLC-UV可成为一种用于植物激素分析的灵敏且经济高效的方法。
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