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.

中文翻译：

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单次免疫亲和柱净化后，对植物样品中 ppb 水平的植物激素脱落酸进行高效液相色谱定量


紫外检测高效液相色谱（HPLC-UV）由于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纯化 (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）的检出限低至1 ng/g。开发了单克隆抗体 3F1 (mAb 3F1) 来特异性识别游离 ABA，但不识别 ABA 类似物。 mAb 3F1 固定化免疫亲和柱的容量为 850 ng/mL，标准品洗脱效率为 88.8–105%。该柱对 ABA 的萃取回收率为 80.4% 至 108.9%。采用 IAC-HPLC-UV 检测各种植物样品中的 ABA 含量。结果通过超高效液相色谱-电喷雾串联质谱法进行验证。 IAC-HPLC-UV 是一种灵敏且经济高效的植物激素分析方法。