Chen Min, Nan Tie-Gui, Xin Jie, Cui Li, Zhang Bo, Wang Xiao, Wang Bao-Min
School of Pharmacy, Linyi University, Linyi 276000, China.
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Int J Anal Chem. 2020 Mar 16;2020:4294826. doi: 10.1155/2020/4294826. eCollection 2020.
Rhein is an important quality-control marker of . The aim of this study was to develop an indirect competitive enzyme-linked immunosorbent assay (icELISA) for rhein detection, which acts as a powerful tool for quality control and proper usage of . First, a specific and sensitive monoclonal antibody (mAb) against rhein was produced from a stable hybridoma cell line, 1F8, generated by the fusion of mouse myeloma sp2/0 with spleen cells obtained from a Bal b/c mouse immunized with rhein-BSA. Then, an icELISA method was developed with an IC value and working range of 0.05 g L and 0.02-0.11 g L, respectively. The icELISA revealed high assay specificity, since it only had a relatively high cross reactivity with aloe-emodin (27%) and almost no cross reactivity with any other anthraquinones (<1%). When spiked with 0.2-2 mg kg of rhein, the recoveries ranged from 84.19% to 102.90%. Finally, icELISA was used to detect rhein contents of collected from different regions, and the results corresponded well with those of HPLC. Overall, the developed icELISA with high specificity and sensitivity provided a rapid and simple method for rhein detection, and it may be a powerful tool for quality control and proper usage of .
大黄酸是……的重要质量控制标志物。本研究的目的是开发一种用于检测大黄酸的间接竞争酶联免疫吸附测定法（icELISA），该方法可作为质量控制和……正确使用的有力工具。首先，从稳定的杂交瘤细胞系1F8产生了一种针对大黄酸的特异性和敏感性单克隆抗体（mAb），该细胞系由小鼠骨髓瘤sp2/0与用大黄酸 - 牛血清白蛋白免疫的Bal b/c小鼠的脾细胞融合而成。然后，开发了一种icELISA方法，其IC值和工作范围分别为0.05 g/L和0.02 - 0.11 g/L。该icELISA显示出高测定特异性，因为它仅与芦荟大黄素具有相对较高的交叉反应性（27%），与任何其他蒽醌几乎没有交叉反应性（<1%）。当添加0.2 - 2 mg/kg的大黄酸时，回收率范围为84.19%至102.90%。最后，使用icELISA检测从不同地区收集的……中大黄酸的含量，结果与高效液相色谱法的结果吻合良好。总体而言，所开发的具有高特异性和敏感性的icELISA为大黄酸检测提供了一种快速简便的方法，并且它可能是质量控制和……正确使用的有力工具。
