Preparation of Room⁃Temperature Phosphorescence Quantum Dots Functionalized with Choline Oxidase and Quantitative Determination of Choline Chloride

doi:10.19894/j.issn.1000-0518.210209

Chinese Journal of Applied Chemistry ›› 2022, Vol. 39 ›› Issue (5): 828-836.DOI: 10.19894/j.issn.1000-0518.210209

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Preparation of Room⁃Temperature Phosphorescence Quantum Dots Functionalized with Choline Oxidase and Quantitative Determination of Choline Chloride

Jin-Zhi LYU(), Xin-Hao ZHANG

School of Life Science，Shanxi Normal University，Linfen 041004，China

Received:2021-04-27 Accepted:2021-09-26 Published:2022-05-01 Online:2022-05-24
Contact: Jin-Zhi LYU
About author:lvjinzhi208@126.com
Supported by:
the National Natural Science Foundation of China(32001202)

Abstract

Abstract:

Environment-friendly nanobiosensors can improve the detection performance of traditional biomolecular sensors and are of importance in practical applications. In this study， a kind of room temperature phosphorescence （RTP） quantum dots （QDs）（ChOx RTP QDs） nanobiosensor was prepared by mineralization at room temperature （25 ℃）， using choline oxidase （ChOx） as the template. The specific enzyme substrate reaction between ChOx and choline oxidase and the photoinduced electron transfer （PIET） were used to detect the RTP of choline oxidase. The linear range of chox detection is 0.05~20 mmol/L， and the detection limit is 0.02 μmol/L. This method is based on the RTP property of QDs， which can effectively avoid the interference of background fluorescence of biological samples and does not need complex sample pretreatment process. Therefore， this method is more suitable for the quantitative detection of choline oxidase in biological samples.

Key words: Choline oxidase, Room temperature phosphorescent quantum dots, Choline chloride

CLC Number:

O655

Jin-Zhi LYU, Xin-Hao ZHANG. Preparation of Room⁃Temperature Phosphorescence Quantum Dots Functionalized with Choline Oxidase and Quantitative Determination of Choline Chloride[J]. Chinese Journal of Applied Chemistry, 2022, 39(5): 828-836.

Figures/Tables 7

Fig.1 （A） Synthesis process and diagram of ChOx RTP QDs；（B） Detection of choline oxidase by ChOx RTP QDs

Fig.2 （A） Relation between n（Mn2+）∶n（Mn2++Zn2+） and RTP intensity of ChOx RTP QDs；（B） Relation between n（S2-）/n（Mn2++Zn2+）and RTP intensity of ChOx RTP QDs；（C） Relation between pH and RTP intensity of ChOx RTP QDs；（D） Relation between ChOx concentration and RTP intensity of ChOx RTP QDs. （λex=295 nm，λem=585 nm）

Fig.3 （A） UV spectra of MPA-capped Mn-ZnS QDs （a）， ChOx RTP QDs （b） and ChOx （c）；（B） Maximum excitation peak （a） and maximum emission peak （b） of ChOx RTP QDs；（C） Phosphorescence lifetime of ChOx RTP QDs；（D） TEM image of ChOx RTP QDs

Fig.4 （A） Variation spectra of RTP intensity of ChOx RTP QDs with the increase of choline chloride concentration （0， 0.05， 0.1， 0.2， 0.5， 1.0， 2.0， 10， 20， 30 and 40 mmol/L）. Insets： plot of ΔRTP/RTP0vs.reciprocal concentration and linear fitting；（B） Standard curve of choline chloride detection based on ChOx RTP QDs；（C） Influences of pH on ΔRTP/RTP0；（D） Influences of temperature on ΔRTP/RTP0

Fig.5 CD spectra of ChOx before （a） and after （b） fixation

Table 1 Effect of co?existing substance on the RTP intensity of 0.1 mmol/L choline

共存物质 Co?existing substance	共存物质浓度/氯化胆碱浓度 c（Co?existing substance）/c（choline）	RTP变化率 Change of the RTP intensity/%
钾离子 K⁺	80	-4.7
钠离子 Na⁺	300	2.6
钙离子 Ca²⁺	10	-4.2
镁离子 Mg²⁺	8	3.9
锌离子 Zn²⁺	0.06	4.9
亚铁离子 Fe²⁺	0.0004	-4.6
锰离子 Mn²⁺	0.005	-2.7
葡萄糖 Glucose	80	-5.3
丙氨酸 Alanine	6	-2.9
脯氨酸 Proline	0.05	5.6
赖氨酸 Lysine	0.05	-3.1
谷胱甘肽 Glutathione	0.05	-4.1
半胱氨酸 Cysteine	0.0005	2.8

Table 2 Recovery for the determination of choline chloride in real samples （n = 3）

样品

Sample

样品中氯化胆碱浓度

Concentration of choline chloride in the sample

c/（mmol·L^-1）

氯化胆碱添加浓度

Choline chloride addition concentration

c/（mmol·L^-1）

检测浓度

Detection concentration

c/（mmol·L^-1）

回收率

Rate of recovery/%

人尿液

Urine

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Preparation of Room⁃Temperature Phosphorescence Quantum Dots Functionalized with Choline Oxidase and Quantitative Determination of Choline Chloride

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