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改进的具有增强稳定性的细胞膜生物亲和性样品预处理技术,用于筛选中药注射剂中的潜在致敏成分。

Improved cell membrane bioaffinity sample pretreatment technique with enhanced stability for screening of potential allergenic components from traditional Chinese medicine injections.

作者信息

Bu Yusi, Hu Qi, Xu Ke, Xie Xiaoyu, Wang Sicen

机构信息

School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, China.

出版信息

J Mater Chem B. 2018 Jan 28;6(4):624-633. doi: 10.1039/c7tb02768k. Epub 2018 Jan 10.

DOI:10.1039/c7tb02768k
PMID:32254491
Abstract

Aiming at improving reliability and tedious analysis time in conventional cell membrane chromatography, an improved bioaffinity sample pretreatment technique with enhanced stability was developed to fast screen and extract potential allergenic components from traditional Chinese medicine injections. In this study, rat basophilic leukemia-2H3 cell membrane coated silica particles (RBL-2H3/CMCSPs) were fabricated by irreversible adsorption between the cell membrane and silica and self-fusion of the cell membrane, which could simulate drug-receptor interactions in vitro. Also, benefiting from the use of paraformaldehyde, the average recoveries of the six batches of RBL/CMCSPs were 90.2% with a relative standard deviation of less than 7.8%, showing that the stability of the materials was remarkably improved compared to materials without fixation. After the successful characterization by spectroscopic and imaging instruments, the prepared RBL-2H3/CMCSPs exhibited desirable adsorption capacity and selectivity. The RBL-2H3/CMCSPs combined with high performance liquid chromatography coupled with time of flight mass spectrometry were then successfully applied to screen and identify two potential allergenic components from huangqi injection, the allergenic activities of which were further investigated by β-hexosaminidase release assay and histamine release assay in vitro. Overall, this work provides a good platform for the fabrication of bioaffinity sample pretreatment materials with high stability and a long lifespan, which can be a time-saving and energy-saving bioaffinity method to rapidly screen and preconcentrate target compounds from complex samples.

摘要

针对传统细胞膜色谱中可靠性低和分析时间冗长的问题,开发了一种稳定性增强的改进型生物亲和样品预处理技术,用于快速筛选和提取中药注射剂中的潜在致敏成分。在本研究中,通过细胞膜与二氧化硅之间的不可逆吸附以及细胞膜的自融合制备了大鼠嗜碱性白血病-2H3细胞膜包被的二氧化硅颗粒(RBL-2H3/CMCSPs),其可在体外模拟药物-受体相互作用。此外,得益于多聚甲醛的使用,六批RBL/CMCSPs的平均回收率为90.2%,相对标准偏差小于7.8%,表明与未固定的材料相比,该材料的稳定性显著提高。经光谱和成像仪器成功表征后,制备的RBL-2H3/CMCSPs表现出理想的吸附容量和选择性。然后,将RBL-2H3/CMCSPs与高效液相色谱-飞行时间质谱联用,成功应用于筛选和鉴定黄芪注射液中的两种潜在致敏成分,并通过体外β-己糖胺酶释放试验和组胺释放试验进一步研究了它们的致敏活性。总体而言,这项工作为制备具有高稳定性和长寿命的生物亲和样品预处理材料提供了一个良好的平台,这可能是一种省时省力的生物亲和方法,可从复杂样品中快速筛选和预富集目标化合物。

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Pharmacol Ther. 2020 Dec;216:107680. doi: 10.1016/j.pharmthera.2020.107680. Epub 2020 Sep 18.