狼毒乙素分子印迹膜荧光传感器的制备及在中药材检测中的应用

doi:10.7503/cjcu20180814

摘要/Abstract

摘要：

以狼毒大戟根部提取的狼毒乙素(ECB)为模板分子, 7-二乙氨基香豆素-3-羧酸与3-溴丙烯缩合生成的烯丙端基香豆素类化合物(DCAC)为荧光单体, 丙烯酰胺为功能单体, 乙二醇二甲基丙烯酸酯为交联剂, 偶氮二异丁腈为引发剂, 通过自由基聚合制备了具有强烈青色荧光, 并且可以选择性富集狼毒乙素的分子印迹膜荧光传感器(FL-MIF). 研究结果表明, FL-MIF吸附ECB的线性范围为1.00~50.00 μmol/L, 在线性范围内, 随着ECB浓度递增, 青色荧光(489 nm)呈现出明显的荧光猝灭响应, 狼毒乙素的检测限为0.29 μmol/L. FL-MIF对ECB具有良好的特异选择性识别性能, 印迹因子为3.58, 明显高于类似结构的2-甲氧基-4-羟基苯甲酸(MHBA)、没食子酸(GA)和3,3'-二乙酰基-4,4'-二甲氧基-2,2',6,6'-四羟基二苯基甲烷(DDTPM). 重复再生性能优异的FL-MIF可应用于实际样品中药材狼毒大戟ECB含量的检测.

关键词: 狼毒乙素, 分子印迹膜, 香豆素, 荧光传感器

Abstract:

Using Ebracteolata compound B(ECB) extracted from the root of Euphorbia Fischeriana steud as template molecule, 7-diethylamine coumarin-3-allyl terminal compounds(DCAC) synthesized by condensation reaction between 7-diethylamine coumarin-3-carboxylic acid and 3-bromopropene as fluorescent monomer, acrylamide as the functional monomer, ethylene glycol dimethacrylate as the crosslinking agent, azodiisobutyronitrile as the initiator, the molecularly imprinted fluorescent film sensor with strong cyan fluorescence was achieved through the free radical polymerization, which could selectively assemble ECB. The adsorption performance of the film showed that the linear range of ECB was 1.00—50.00 μmol/L. The cyan fluorescence(489 nm) quenching response of the film was shown obviously with the increasing concentration of ECB. The sensor could be used to detect the low concentration of ECB, with a detection limit to 0.29 μmol/L. The imprinting factor was 3.58, obviously higher than the analogues(2-methoxy-4-hydroxybenzoic acid, gallic acid and 3,3'-diacetyl-4,4'-dimethoxy-2,2',6,6'-tetrahydroxydiphenylmethane). The sensor with excellent reusability has been successfully applied to detect the content of ECB in Euphorbia fischeriana Steud.

Key words: Ebracteolata compound B, Molecularly imprinted film, Coumarin, Fluorescenc sensor

中图分类号:

O658

TrendMD:

马玉坤, 王海君, 郭梦岩. 狼毒乙素分子印迹膜荧光传感器的制备及在中药材检测中的应用. 高等学校化学学报, 2019, 40(7): 1381.

MA Yukun, WANG Haijun, GUO Mengyan. Preparation of ECB Molecularly Imprinted Film Fluorescent Sensor for Detection of Traditional Chinese Herbal Medicine^†. Chem. J. Chinese Universities, 2019, 40(7): 1381.

图/表 16

Scheme 1 Synthetic routes of fluorescent monomer DCAC

Scheme 2 Structures of ECB and three other similar compoundsECB: Ebracteolata Compound B; MHBA: 2-hydroxy-6-methoxyacetopheonone; GA: 3,5-trithydroxybenzoic acid; DDTPM: 3,3’-diacetyl-4,4’-dimethoxy-2,2’,6,6’-tetrahydroxydiphenylmethane.

Fig.1 Ultrahigh performance liquid chromatography of ECB

Scheme 3 Synthesis of FL-MIF sensor and the proposed quenching mechanism on the basis of energy transfer(ET)

Fig.2 FTIR spectra of FL-MIF(a), FL-NIF(b), DCAC(c) and ECB(d)

Fig.3 SEM images of FL-MIF with different manification

Fig.4 Fluorescence emission spectra of samples a. DCAC; b. DACCA; c. FL-NIF; d. FL-MIF, before washing; e. FL-MIF, after washing.

Fig.5 Dynamic adsorption curves of FL-MIF(a) and FL-NIF(b) toward ECB

Fig.6 Static adsorption curves of FL-MIF(a) and FL-NIF(b) toward ECB

Fig.7 Fluorescence responses of FL-MIF(A) and FL-NIF(B) with the increasing concentration of ECBInsets: Stern-Volmer curves.

Fig.8 Stern-Volmer curves of FL-MIF(A) and FL-NIF(B) for analogues

Fig.9 Stern-Volmer quenching constant diagram of FL-MIF and FL-NIF for analogues

Fig.10 Fluorescence intensity contrast diagram of FL-MIF for ECB with different ratio of competitors

Fig.11 Regeneration of FL-MIF

Fig.12 Fluorescence spectra of FL-MIF with different content of ECB in sample 1cstandard/(μmol·L-1): a. 0; b. 5; c. 10; d. 15.

Table 1 Detection results of the ECB contents in Euphorbia fischeriana Steud from different habitats

No.	Source	ECB content/(μmol·g^-1)
No.	Source	FL-MIF	HPLC
1	Medicine Garden of Qiqihar Medical University	1.09	1.11
2	Jiefang Township, Yi’an County, Qiqihar	1.85	1.81
3	Dajiazi Mountain, Nianzishan District, Qiqihar	1.29	1.30
4	Green grassland, Dulbert County, Daqing	1.92	1.91
5	Chabaqi Township, Arong Banner, Hulunbuir	2.44	2.47

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