Analysis of Imidazole Propionic Acid in Serum of Patients with Type 2 Diabetes Based on NiO@SiO2 Solid-phase Extraction Coupled with Liquid Chromatography-Mass Spectrometry †

doi:10.7503/cjcu20190483

Abstract

Abstract:

Imidazole propionic acid can cause insulin resistance in human through mammalian target of rapamycin complex 1(mTORC1) pathway, leading to type 2 diabetes(T2D). Therefore, the accurate quantification of imidazole propionic acid benefits for the diagnosis of type 2 diabetes. Based on the coordination between NiO and imidazole groups, NiO@SiO₂ was used for the selective extraction of imidazole propionic acid in serum. Firstly, the NiO@SiO₂ solid phase extraction(SPE) conditions were optimized as follows: the amount of absorbent was 200 mg, the loading solution was 20 mmol/L of phosphate buffer(pH=3.0) and 1.0 mL of aqueous solution containing 1% NH₃·H₂O was used as the elution. Then, a simple and effective method for the analysis of imidazole propionic acid in serum was established by coupling NiO@SiO₂ solid phase extraction with high performance liquid chromatography-mass spectrometry(HPLC-MS). The linearity of the developed method was in the range of 0.05—10 ng/mL with a good linearity(R ²≥0.996). The LOD and LOQ were 0.02 and 0.05 ng/mL, respectively. The recovery was 84.0%—119%, and the relative standard deviation(RSD) was less than 17.2%. The established method was applied to the detection of imidazole propionic acid in patients with type 2 diabetes and normal subjects. There was a significant difference in imidazole propionic acid content between type 2 diabetic patients and normal subjects, indicating that accurate quantification of imidazole propionic acid has great potential in medical diagnosis.

Key words: Imidazolepropionic acid, NiO@SiO₂, Type 2 diabetes, Solid phase extraction(SPE), High performance liquid chromatography-mass spectrometry(HPLC-MS)

CLC Number:

O657.63

TrendMD:

WANG Tianqi,YU Qiongwei,FENG Yuqi. Analysis of Imidazole Propionic Acid in Serum of Patients with Type 2 Diabetes Based on NiO@SiO₂ Solid-phase Extraction Coupled with Liquid Chromatography-Mass Spectrometry ^†[J]. Chem. J. Chinese Universities, 2020, 41(2): 262.

Figures/Tables 4

Fig.1 Optimization of the extraction conditions (A) pH of sample matrix(3, 5, 7, 9 and 11); (B) NiO@SiO2 sorbent amount(10, 20, 50, 100, 150 and 200 mg); (C) six kinds of eluent solution: a. ammonium hydroxide/acetonitrile(1:99, volume ratio); b. ammonium hydroxide/H2O(1:99, volume ratio); c. ammonium hydroxide/H2O(5:95, volume ratio); d. formic acid/H2O(1:1000, volume ratio); e. formic acid/acetonitrile(1:1000, volume ratio); f. H2O. (D) Eluent solution: ammonium hydroxide/H2O(1:99, volume ratio).

Fig.2 Chromatogram of serum and spiked serum with 2.5 ng/mL imidazole propionic acid after NiO@SiO2 SPE(A) and the mass spectrum of the chromatographic peak at 6.89 min(B)

Analyte	Precision(RSD, %)						Recovery(%, n=5)
	Intra-day(n=5)			Inter-day(n=3)			Recovery(%, n=5)
	0.25 ng/mL	1.0 ng/mL	10 ng/mL	0.25 ng/mL	1.0 ng/mL	10 ng/mL	0.25 ng/mL	1.0 ng/mL	10 ng/mL
Imidazole propionic acid	17.2	1.20	14.3	2.40	2.40	11.8	84.0	119	95.1

No.	ρ/(ng·mL^-1)		No.	ρ/(ng·mL^-1)
No.	T2D group	Control group	No.	T2D group	Control group
1	6.0	8.1	11	5.5	3.1
2	135.6	11.5	12	6.3	2.6
3	12.1	11.1	13	3.3	2.0
4	8.7	4.4	14	2.6	6.3
5	5.8	3.1	15	2.9	1.8
6	3.5	4.2	16	2.3	2.4
7	6.7	12.5	17	3.3	4.0
8	6.9	3.5	18	69.9	7.8
9	10.5	1.5	19	17.2	3.6
10	5.2	3.3	20	10.4	6.0

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Analysis of Imidazole Propionic Acid in Serum of Patients with Type 2 Diabetes Based on NiO@SiO₂ Solid-phase Extraction Coupled with Liquid Chromatography-Mass Spectrometry ^†

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