聚丙烯酸-钙(铈)纳米团簇荧光探针用于无机磷定量检测研究

doi:10.15541/jim20230596

摘要/Abstract

摘要：

无机磷主要以磷酸根的形式存在, 在自然界生命循环和现代工业社会中发挥重要作用。磷酸根离子的快速高效定量检测一直是临床生化分析、工业生产和环境污染监测等领域的研究热点。本工作以聚丙烯酸(PAA)为络合剂, 通过与Ca²⁺和Ce³⁺的络合反应, 合成了具有良好分散性和稳定性的PAA-Ca(Ce)纳米团簇荧光探针。利用298 nm激发光照射该探针与磷酸根的反应产物, 建立其在352 nm处发射峰峰值强度与磷酸根浓度的线性关系图。实验结果表明, (Ca²⁺+Ce³⁺)浓度为37.575 mmol/L的纳米荧光探针与磷酸根浓度的线性关系式为y=1.09x+2.05, 荧光强度可靠性范围为13.5~66.91 mmol/L。与钼锑抗分光光度法测试磷酸根对比, 该方法具有更高的回收率。大鼠血清无机磷检测实验验证了该方法的可靠性。以上结果表明本研究所合成的荧光探针具有良好的磷酸根定量检测性能。

关键词: 稀土离子, 荧光探针, 无机磷, 检测

Abstract:

Phosphorus is one of the important elements in the ecosystem and plays a vital role in the process of life cycle. Inorganic phosphorus is a major form of phosphorus, usually in the form of phosphate ions. The rapid and efficient quantitative detection of phosphate ions has always been a hot research direction in the fields of clinical chemistry, pharmacology, biochemical analysis, industrial production and environmental pollution monitoring, but still facing some challenges in accuracy and convenient in some special circumstances. In present work, PAA-Ca(Ce) nanocluster fluorescent probes with good dispersibility and stability were synthesized by complexing with Ca²⁺ and Ce³⁺ using polyacrylic acid (PAA) as complexing agent. The reaction product between the probe and the phosphate ion was irradiated by 298 nm excitation light, and a linear graph was established based on the correlation between the peak intensity at the emission peak of 352 nm and the phosphate ion concentration. The experimental results show that the linear relation of the nanofluorescent probe with the (Ca²⁺+Ce³⁺) concentration of 37.575 mmol/L is y=1.09x+2.05, and the reliability range of fluorescence intensity is 13.5-66.91 mmol/L. Compared with molybdenum-antimony resistance spectrophotometry, this method has a higher recovery. The experimental results of inorganic phosphorus in rat serum verify the reliability of the method. The results above indicate that the fluorescence probe studied in this paper has good quantitative detection performance of phosphate.

Key words: rare earth ion, fluorescent probe, inorganic phosphorus, detection

中图分类号:

TQ174

陈甲, 范依然, 闫文馨, 韩颖超. 聚丙烯酸-钙(铈)纳米团簇荧光探针用于无机磷定量检测研究[J]. 无机材料学报, 2024, 39(9): 1053-1062.

CHEN Jia, FAN Yiran, YAN Wenxin, HAN Yingchao. Polyacrylate-calcium (cerium) Nanocluster Fluorescent Probes for Quantitative Detection of Inorganic Phosphorus[J]. Journal of Inorganic Materials, 2024, 39(9): 1053-1062.

图/表 10

图1 PAA-Ca(Ce)纳米荧光探针的形貌尺寸和化学结构

Fig. 1 Morphology, size and structure of PAA-Ca(Ce) nanofluorescent probe (a) Cryo-TEM images; (b) Particle size statistical distribution calculated from cryo-TEM images; (c) DLS particle size distribution; (d) FT-IR spectra of the products of PAA-Ca(Ce) nanoprobes with PO43- solutions at concentration of 0 (I), 10 (II) and 50 (III) mmol/L

图2 PAA-Ca(Ce)纳米荧光探针检测PO43-的荧光稳定性

Fig. 2 Fluorescence stability of PAA-Ca(Ce) nanofluorescent probe for PO43- (a) Fluorescence emission spectra of PAA-Ca(Ce) and PO43- after reaction within 7 days; (b) Fluorescence emission spectra of the original PAA-Ca(Ce) solution and the cold-dry redissolved solution reacting with PO43-; (c) Fluorescence emission spectra of PAA-Ca(Ce) solution reacting with PO43- before and after filtration by ϕ0.22 µm membrane

图3 不同(Ca2++Ce3+)浓度下PAA-Ca(Ce)荧光探针与PO43-反应产物的荧光发射光谱图, 以及PO43-浓度-荧光强度线性关系

Fig. 3 Fluorescence emission spectra of the reaction products between PAA-Ca(Ce) fluorescent probes and PO43- with different concentrations of (Ca2++Ce3+), and corresponding linear relationship of PO43- concentration-fluorescence intensity (a-c) Fluorescence emission spectra of PAA-Ca(Ce) probe and PO43- with different concentrations excited at 298 nm excitation under the conditions of (Ca2++Ce3+) concentration of (a) 18.7875, (b) 35.575 and (c) 75.150 mmol/L, respectively; (d-f) Relationship curves and linear fitting relationships between fluorescence peak intensity at 352 nm and PO43- concentration under the conditions of (Ca2++Ce3+) concentration of (d) 18.7878, (e) 35.575 and (f) 75.150 mmol/L, respectively; Colorful figures are available on website

图4 低PO43-浓度下的PO43-浓度-荧光强度关系图

Fig. 4 Relationship between PO43- concentration and fluorescence intensity at low PO43-concentration (a) Fluorescence emission spectra of PAA-Ca(Ce) probe with different concentrations of PO43- at 298 nm excition at (Ca2++Ce3+) concentration of 35.575 mmol/L; (b-d) Relationship between the fluorescence peak intensity at 352 nm and the concentration of PO43- with their corresponding fitting curves derivated from different models; Colorful figures are available on website

图5 反应时间与温度对荧光强度的影响

Fig. 5 Influence of reaction time and temperature on fluorescence intensity (a, b) Changes of fluorescence intensity detected at different time; (d, c) Changes of fluorescence intensity detected at different temperatures; Colorful figures are available on website

表1 荧光探针法和钼锑抗分光光度法测定PO43-的回收率

Table 1 Recovery of PO43- detected by fluorescence probe (FP) method in contrast to that by molybdenum-antimony resistance spectrophotometry (MARSP)

PO₄^3-added / (mmol·L^-1)	FP method		MARSP
PO₄^3-added / (mmol·L^-1)	PO₄^3-detected/ (mmol·L^-1)	Recovery/ %	PO₄^3-detected/ (mmol·L^-1)	Recovery/ %
10	9.633	96.326	9.096	90.962
20	20.611	103.056	18.694	93.472
30	28.856	96.186	26.634	88.780
40	40.009	100.022	37.719	94.299
50	48.634	97.269	47.779	95.559
60	60.304	100.507	55.134	91.891
70	68.462	97.802	66.391	94.844
80	79.159	98.949	75.191	93.989
90	87.964	97.737	87.080	96.755
100	100.526	100.526	94.505	94.505
110	110.024	100.022	99.928	90.844
120	116.614	97.178	115.249	96.041
130	131.590	101.223	122.934	94.564
140	139.348	99.534	125.775	89.839
150	145.318	96.879	140.961	93.974
160	161.331	100.832	156.500	97.813
170	171.033	100.608	164.876	96.986
180	176.630	98.128	170.588	94.771
190	191.582	100.832	178.270	93.826
200	198.966	99.483	196.275	98.138
Average recovery (mean±SD)/%	99.155±1.880		94.093±2.566

图6 仪器响应值与PO43-浓度的关系图

Fig. 6 Relationship between instrument response value and PO43- concentration

图7 不同化学环境因素对PAA-Ca(Ce)纳米荧光探针的影响

Fig. 7 Influence of different chemical environmental factors on PAA-Ca(Ce) nano-fluorescent probes (a, d) Fluorescence emission spectra of PAA-Ca(Ce) after reaction with various substances and comparison of fluorescence intensity at 352 nm emission peak, respectively; (b, e) Fluorescence emission spectra of PAA-Ca(Ce) reacting with PO43- at different concentrations of NaCl (0-100 mmol/L) and comparison of emission peak fluorescence intensity at 352 nm, respectively; (c, f) Fluorescence emission spectra of PO43- detected by PAA-Ca(Ce) under the conditions of PO43- concentration at 10 and 50 mmol/L while medium composed by ammonia-ammonium chloride buffer or ultra-pure water, and the fluorescence intensity comparison diagram of the emission peak at 352 nm; Colorful figures are available on website

表2 大鼠血清中PO43-的测定

Table 2 Determination of PO43- in mouse serum

Sample	FP method	MARSP	p value
1	2.648	2.552	0.9626
2	2.375	2.451
3	2.599	2.604
Average±SD	2.541±0.119	2.536±0.064

图8 荧光探针法检测无机磷的机理示意图

Fig. 8 Schematic of fluorescence probe method for sensing PO43-

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