Key Laboratory of Quality and Safety Control of Citrus Fruits, Ministry of Agriculture and Rural Affairs, Southwest University, Chongqing 400712, China; National Citrus Engineering Research Center, Chongqing 400712, PR China; Laboratory of Quality & Safety Risk Assessment for Citrus Products (Chongqing), Ministry of Agriculture, Citrus Research Institute, Southwest University, Chongqing 400712, China.
Key Laboratory of Quality and Safety Control of Citrus Fruits, Ministry of Agriculture and Rural Affairs, Southwest University, Chongqing 400712, China; National Citrus Engineering Research Center, Chongqing 400712, PR China; Laboratory of Quality & Safety Risk Assessment for Citrus Products (Chongqing), Ministry of Agriculture, Citrus Research Institute, Southwest University, Chongqing 400712, China.
Food Chem. 2024 Jul 30;447:138989. doi: 10.1016/j.foodchem.2024.138989. Epub 2024 Mar 13.
Limonin is an intensely bitter and highly oxidized tetracyclic triterpenoid secondary metabolite, which is abundant in the Rutaceae and Meliaceae, especially in Citrus. In order to detect limonin content in complex substrates such as citrus and traditional Chinese medicine, monoclonal antibodies specifically recognizing limonin were prepared and an indirect competitive enzyme-linked immunosorbent assay (icELISA) was established. The median inhibition concentration (IC) was 5.40 ng/mL and the linear range was 1.25-23.84 ng/mL. The average recoveries from citrus peel and pulp samples were 95.9%-118.8% and 77.5%-113.1%, respectively. Moreover, the contents of limonin in 6 citrus samples and 4 herbal samples were analyzed by icELISA and UPLC-MS, and the results of the two methods were consistent. This validation is sufficient to demonstrate that the developed immunoassay is applicable for the detection of limonin in citrus and herbal samples and has the advantage of high efficiency, sensitivity, and convenience.
柠烯是一种苦味浓烈且高度氧化的四环三萜类次生代谢物，在芸香科和楝科中含量丰富，特别是在柑橘中。为了检测柑橘和中药等复杂基质中的柠烯含量，制备了特异性识别柠烯的单克隆抗体，并建立了间接竞争酶联免疫吸附测定法（icELISA）。该方法的半数抑制浓度（IC）为 5.40ng/mL，线性范围为 1.25-23.84ng/mL。从柑橘果皮和果肉样品中的平均回收率分别为 95.9%-118.8%和 77.5%-113.1%。此外，通过 icELISA 和 UPLC-MS 分析了 6 个柑橘样品和 4 个草药样品中的柠烯含量，两种方法的结果一致。该验证足以证明所开发的免疫测定法适用于柑橘和草药样品中柠烯的检测，具有高效、灵敏和方便的优点。
