Efficient Determination of Alkaline Phosphatase Activity Based on Graphene Quantum Dots Fluorescent Probes

doi:10.7503/cjcu20210048

Abstract

Abstract:

Based on the static fluorescence quenching effects of benzoquinones on graphene quantum dots（GQDs）， a GQDs-based fluorescent probe was established to detect the activity of alkaline phosphatase（ALP）. Under the catalytic action of horseradish peroxidase， hydrogen peroxide（H₂O₂） could generate hydro-xyl radicals（·OH）， and ·OH oxidized o-dihydroxybenzene to produce o-benzoquinone subsequently， resul-ting in the fluorescence quenching of GQDs. ALP catalyzed the production of reductive ascorbic acid by ascorbic acid 2-phosphat. Ascorbic acid efficiently removed H₂O₂ and ·OH from the solution， leading to the inhibition of the production of o-benzoquinone and the fluorescence recovery of GQDs. Therefore， a highly sensitive determination strategy for ALP activity was established. Under the optimal experimental conditions， the detection limit of ALP activity was 0.084 U/L. This method was used to evaluate ALP activity in human serum samples. Such approach can provide theoretical basis for the diagnosis and treatment of ALP-related diseases.

Key words: Graphene quantum dot, Fluorescent probe, Alkaline phosphatase, Static quenching, Enzymatic activity

CLC Number:

O657.3

TrendMD:

HUANG Shan, YAO Jiandong, NING Gan, XIAO Qi, LIU Yi. Efficient Determination of Alkaline Phosphatase Activity Based on Graphene Quantum Dots Fluorescent Probes[J]. Chem. J. Chinese Universities, 2021, 42(8): 2412.

Figures/Tables 16

References 37

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Co?existing substance	Δ（I/I₀）（%）	Co?existing substance	Δ（I/I₀）（%）
L?Aspartic acid	+1.52	Lactose	+2.74
L?Serine	-3.96	Galactose	+3.79
L?Glutamate	-4.93	Maltose	-1.13
L?Lysine	+2.36	Uric acid	-4.42
L?Phenylalanine	-2.51	Cholesterol	-0.95
D?Tryptophan	+4.18	Guanosine	+2.76
L?Tryptophan	-1.61	Adenosine	+3.59
L?Glutathione oxidized	+1.65	AlCl₃	+1.04
L?Glycine	+3.32	NiCl₂	-4.03
L?Arginine	-4.55	MnSO₄	+3.59
L?Methionine	+0.47	MgCl₂	+2.42
L?Glutamine	-1.48	CrCl₃	+1.39
L?Leucine	1.48	CdCl₂	-4.29
L?Valine	+1.36	CaCl₂	-4.56
L?α?Alanine	+4.67	NaCl	+3.80

Co?existing substance	Δ（I/I₀）（%）	Co?existing substance	Δ（I/I₀）（%）
L?Aspartic acid	+1.52	Lactose	+2.74
L?Serine	-3.96	Galactose	+3.79
L?Glutamate	-4.93	Maltose	-1.13
L?Lysine	+2.36	Uric acid	-4.42
L?Phenylalanine	-2.51	Cholesterol	-0.95
D?Tryptophan	+4.18	Guanosine	+2.76
L?Tryptophan	-1.61	Adenosine	+3.59
L?Glutathione oxidized	+1.65	AlCl₃	+1.04
L?Glycine	+3.32	NiCl₂	-4.03
L?Arginine	-4.55	MnSO₄	+3.59
L?Methionine	+0.47	MgCl₂	+2.42
L?Glutamine	-1.48	CrCl₃	+1.39
L?Leucine	1.48	CdCl₂	-4.29
L?Valine	+1.36	CaCl₂	-4.56
L?α?Alanine	+4.67	NaCl	+3.80

Sample	Spiked/（U·L^-1）	Found/（U·L^-1）	Recovery（%）	RSD（%）
Human serum sample 1	0	1.39—1.45	—	—
	0.5	1.90—1.92	96.5—100.4	2.62
	3	4.42—4.52	99.1—103.5	3.15
	5	6.31—6.36	97.9—98.9	0.02
Human serum sample 2	0	1.16—1.23	—	—
	0.5	1.69—1.72	98.5—102.8	0.69
	3	4.12—4.27	97.5—103.4	2.28
	5	5.98—6.12	95.5—98.4	2.03
Human serum sample 3	0	0.87—0.94	—	—
	0.5	1.39—1.42	97.9—103.7	4.09
	3	3.90—4.04	100.8—103.4	0.46
	5	5.99—6.03	101.7—102.4	0.59

Sample	Spiked/（U·L^-1）	Found/（U·L^-1）	Recovery（%）	RSD（%）
Human serum sample 1	0	1.39—1.45	—	—
	0.5	1.90—1.92	96.5—100.4	2.62
	3	4.42—4.52	99.1—103.5	3.15
	5	6.31—6.36	97.9—98.9	0.02
Human serum sample 2	0	1.16—1.23	—	—
	0.5	1.69—1.72	98.5—102.8	0.69
	3	4.12—4.27	97.5—103.4	2.28
	5	5.98—6.12	95.5—98.4	2.03
Human serum sample 3	0	0.87—0.94	—	—
	0.5	1.39—1.42	97.9—103.7	4.09
	3	3.90—4.04	100.8—103.4	0.46
	5	5.99—6.03	101.7—102.4	0.59

Method	Probe	Linear range/（U·L^-1）	Detection limit/ （U·L^-1）	Samples detection	Ref.
Fluorimetry	Carbon quantum dots	3.4—100	0.9	—	［35］
Electrochemical	CdS@graphene?CoOOH nanosheets	10—300	1.5	Human serum samples	［17］
Fluorimetry	Polydopamine/MnO₂ nanocomposites	1—80	0.34	Human serum samples	［20］
Fluorimetry	Hemicyanine?based near?infrared probe	10—2000	3	Cell， tissue and living animal	［36］
Colorimetric	Cu（II）?phenanthroline complex	0—200	1.25	Human serum samples	［19］
Fluorimetry	Silver nanocluster	30—3000	5	Bovine serum samples	［23］
Fluorimetry	WS₂ quantum dots	0.5—10	0.2	Human serum samples	［25］
Fluorimetry	Polydopamine nanodots	1—50	0.94	Human serum samples	［21］
Fluorimetry	Protein stabilized gold nanocubes	—	1.616	Human serum samples	［24］
Fluorimetry	Electrospun fibrous strips	0—80	1.5	Human serum samples	［37］
Fluorimetry	Carbon quantum dots	4.6—383.3	1.4	—	［26］
Fluorimetry	GQDs	0.1—5	0.084	Human serum samples	This work