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基于单克隆抗体的酶联免疫吸附测定法用于定量测定大戟苷R2,作为宁顺和莱州人参的鉴定标志物。

Monoclonal antibody-based enzyme-linked immunosorbent assay for quantification of majonoside R2 as an authentication marker for Nngoc Linh and Lai Chau ginsengs.

作者信息

Chaingam Jiranan, Van Huy Le, Noguchi Kanta, Nuntawong Poomraphie, Vimolmangkang Sornkanok, Yodsurang Varalee, Yusakul Gorawit, Morimoto Satoshi, Sakamoto Seiichi

机构信息

Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.

Department of Biochemistry and Medicinal Chemistry, Research Institute for Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Viet Nam.

出版信息

J Ginseng Res. 2024 Sep;48(5):474-480. doi: 10.1016/j.jgr.2024.05.004. Epub 2024 May 23.

Abstract

BACKGROUND

Recent years have witnessed increasing interest in the high amount of ocotillol-type saponin in , particularly in relation to majonoside R2 (MR2). This unique 3%-5% MR2 content impart Ngoc Linh and Lai Chau ginsengs with unique pharmacological activities. However, in the commercial domain, unauthentic species have infiltrated and significantly hindered access to the authentic, efficacious variety. Thus, suitable analytical techniques for distinguishing authentic Vietnamese ginseng species from others is becoming increasingly crucial. Therefore, MR2 is attracting considerable attention as a target requiring effective management measures.

METHODS

An enzyme-linked immunosorbent assay (ELISA) was developed by producing monoclonal antibodies against MR2 (mAb 16E11). The method was thoroughly validated, and the potential of the immunoassay was confirmed by high-performance liquid chromatography with ultraviolet spectroscopy. Furthermore, ELISA was applied to the assessment of the MR2 concentrations of various spp., including Korean, American, and Japanese ginsengs.

RESULTS AND CONCLUSIONS

An icELISA using mAb 16E11 exhibited linearity between 3.91 and 250 ng/mL of MR2, with detection and quantification limits of 1.53 and 2.50 46.6 ng/mL, respectively. Based on this study, the developed icELISA using mAb 16E11 could be a valuable tool for analyzing MR2 level to distinguish authentic Ngoc Linh and Lai Chau ginsengs from unauthentic ones. Furthermore, the analysis of the samples demonstrated that Ngoc Linh and Lai Chau ginsengs exhibit a notably higher MR2 value than all other spp. Thus, MR2 might be their ideal marker compound, and various bioactivities of this species should be explored.

摘要

背景

近年来,人们对三七中高含量的人参二醇型皂苷,尤其是人参皂苷R2(MR2)的关注度日益增加。这种独特的3%-5%的MR2含量赋予了越南的宁平和莱州人参独特的药理活性。然而,在商业领域,伪品已经混入,严重阻碍了获取正宗、有效的品种。因此,合适的分析技术来区分正宗的越南人参品种与其他品种变得越来越关键。因此,MR2作为一个需要有效管理措施的目标正受到相当大的关注。

方法

通过制备抗MR2的单克隆抗体(mAb 16E11)开发了一种酶联免疫吸附测定(ELISA)方法。该方法经过了全面验证,免疫测定的潜力通过高效液相色谱-紫外光谱法得到了证实。此外,ELISA被应用于评估各种人参属植物中MR2的浓度,包括高丽参、西洋参和日本参。

结果与结论

使用mAb 16E11的间接竞争ELISA在MR2浓度为3.91至250 ng/mL之间呈现线性关系,检测限和定量限分别为1.53和2.50 ng/mL。基于本研究,所开发的使用mAb 16E11的间接竞争ELISA可能是分析MR2水平以区分正宗的宁平和莱州人参与伪品的有价值工具。此外,对样品的分析表明,宁平和莱州人参的MR2值明显高于所有其他人参属植物。因此,MR2可能是它们理想的标记化合物,应该探索该品种的各种生物活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f402/11385409/c0731dd60005/ga1.jpg

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