钌-生物质碳人工酶的制备及在比色检测杀虫剂毒死蜱残留中的应用

doi:10.15541/jim20210365

摘要/Abstract

摘要：

金属纳米材料人工酶以其高稳定性和低成本的特点而被广泛用于生物传感与催化领域。本研究采用[1-甲基-3- 乙基咪唑][二氰胺]离子液体([EMIM][DCA])与氯化钌反应形成[EMIM]₃[Ru(DCA)₆]离子液体, 将蚕丝溶解于[EMIM]₃[Ru(DCA)₆]后经高温碳化得到钌-生物质碳纳米材料。结果显示, 钌-生物质碳纳米材料具有优异的分散性, 平均直径为7.5 nm的钌纳米粒子可均匀地分散在碳片表面, 表现出较强的类氧化酶活性。基于杀虫剂毒死蜱对钌-生物质碳酶活性的抑制作用, 建立了一种测定毒死蜱的比色方法。钌-生物质碳酶能催化3,3′,5,5′-四甲基联苯胺氧化生成蓝色产物。毒死蜱能抑制钌-生物质碳人工酶的活性, 从而导致蓝色产物的吸光度下降。当毒死蜱浓度在10~80 ng/mL之间, 反应体系的吸光度随毒死蜱浓度增大而线性下降, 检出下限为6.5 ng/mL。本方法的灵敏度和稳定性明显优于现有技术, 已成功应用于桃果实中毒死蜱农药残留的快速测定。

关键词: 金属离子液体, 钌纳米粒子, 类氧化酶, 有机磷杀虫剂, 比色检测

Abstract:

Metal nanomaterials as mimic enzyme have been widely used for biosensor and catalysis due to their high stability and low cost. Here, [1-methyl-3-ethylimidazole][dicyandiamide] ionic liquid ([EMIM][DCA]) reacted with ruthenium chloride to form [EMIM]₃[Ru(DCA)₆] ionic liquid. Silk was dissolved in the above ionic liquid and then carbonized at high temperature to obtain Ru-biomass carbon composite. The results show that Ru-biomass carbon offers excellent dispersion, small Ru nanoparticles with an average diameter of 7.5 nm which were uniformly dispersed on the surface of carbon sheets, and high oxidase-like activity. Based on the inhibition of chlorpyrifos on catalytic activity of Ru-biomass carbon, a method was established for colorimetric determination of chlorpyrifos. Chlorpyrifos obviously inhibits the catalytic activity of Ru-biochar carbon, leading to a reduced absorbance of blue product. When the concentration of chlorpyrifos was in the range from 10 to 80 ng/mL, the peak absorbance decreases linearly with the increase of chlorpyrifos concentration, giving a detection limit of about 6.5 ng/mL. The as-proposed method provides higher sensitivity and stability compared with present methods, and has successfully applied to detection of chlorpyrifos in peach samples.

Key words: metal ionic liquid, Ru nanomaterial, oxidase-like enzyme, organophosphorus insecticides, colorimetric detection

中图分类号:

TQ174

曹志军, 李在均. 钌-生物质碳人工酶的制备及在比色检测杀虫剂毒死蜱残留中的应用[J]. 无机材料学报, 2022, 37(5): 554-560.

CAO Zhijun, LI Zaijun. Ruthenium-biocarbon Mimic Enzyme: Synthesis and Application in Colorimetric Detection of Pesticide Chlorpyrifos[J]. Journal of Inorganic Materials, 2022, 37(5): 554-560.

图/表 8

图1 钌-生物质碳复合物的SEM(A)、TEM(B)、HRTEM(C)照片及钌(D)和氮(E)的元素映像图和XRD图谱(F)

Fig. 1 SEM (A), TEM (B), HRTEM images (C), and elemental mappings of Ru (D) and N (E) elements and XRD pattern (F) of the as-prepared Ru-biomass carbon composite

图2 钌-生物质碳复合物的XPS总谱(A)、高分辨C1s(B)、Ru3p(C)和N1s(D)XPS分谱

Fig. 2 Total XPS spectrum (A), high-resolution C1s (B), Ru3p (C) and N1s (D) XPS spectra of Ru-biomass carbon composite

图3 钌-生物质碳复合物(a)、CuO/Pt复合物(b)、钌纳米粒子(c)和生物质碳(d)催化体系的UV-Vis吸收光谱

Fig. 3 Absorption spectra of the reaction system catalysis by Ru-biomass carbon composite (a), CuO/Pt composite (b), Ru nanoparticles (c), and biomass carbon (d)

图4 钌-生物质碳复合物催化在氮气、空气和氧气饱和的TMB溶液中的665 nm波长处的吸光度

Fig. 4 Absorbances at 665 nm of the N2, air and O2 saturated TMB solutions with catalysis by Ru-biomass carbon composite

图5 介质pH(A)、反应时间(B)、反应温度(C)和催化剂用量(D)与反应体系吸光度的关系曲线

Fig. 5 Relationship curves of pH (A), reaction time (B), reaction temperature (C), and catalyst amount (D) with the absorbance of the reaction system

图6 在0、10、20、30、40、50、60、70和80 ng/mL毒死蜱存在下反应体系的吸收光谱(A)和665 nm处的吸光度与毒死蜱浓度的关系曲线(B)

Fig. 6 Absorption spectra of the reaction system in the presence of 0, 10, 20, 30, 40, 50, 60, 70 and 80 ng/mL chlorpyrifos (A) and calibration plot of the absorbance at 665 nm varied with the chlorpyrifos concentration (B)

表1 不同纳米酶比色法检测有机磷农药的比较

Table 1 Comparison of colorimetric methods with different nanozyme materials for detection of organophosphorus pesticides

Chromogenic system	Analysis time/min	Linear range/(ng·mL^-1)	Detection limit/(ng·mL^-1)	Ref.
Gold nanoclusters	40	1500-45000	6000	[23]
Acetylcholine	55	10000-140000	4000	[24]
Cu-Ag/rGO	30	300-9000	1080	[25]
Hemin regulated by oligonucleotide	62	2000-100000	600	[26]
CuO/Pt	50	300-180000	238.8	[19]
Fe-N-C	76	100-10000	97	[27]
Cu NPs	10	300-3000	20.4	[28]
Ru-biomass carbon composite	20	5-80	6.57	This work

表2 桃样品中毒死蜱检测结果

Table 2 Results for detection of chlorpyrifos in peach (n=5)

Sample	Chlorpyrifos added /(ng·mL^-1)	Chlorpyrifos detected by proposed method/(ng·mL^-1)	Chlorpyrifos founded by GC-MS/(ng·mL^-1)	Recovery/%
Honey peach	0	(5.9±0.15)	(6.02±0.12)
Honey peach	10.0	(16.2±0.18)	(16.05±0.10)	101.8
Nectarine	0	(9.98±0.07)	(9.95±0.07)
Nectarine	10.0	(19.91±0.27)	(20.1±0.27)	99.6
Peento	0	(4.96±0.11)	(5.07±0.05)
Peento	10.0	(15.2±0.19)	(15.07±0.25)	101.6

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