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Method for rapidly, sensitively and quantitatively detecting glucose in serum

A technology for quantitative detection of glucose, applied in the field of quantitative detection of glucose in serum, can solve the problems of difficult quantitative detection of glucose, inability to achieve high throughput, matrix signal interference, etc., to enhance selectivity and sensitivity, solve poor reproducibility, Effect of Rapid Quantitative Analysis

Inactive Publication Date: 2017-08-22
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In recent years, glucose sensors based on electrochemical and optical technologies have been widely used due to their low cost; however, such technologies have the disadvantages of signal interference and low sensitivity
Based on this, with the advantages of high accuracy and high sensitivity, the method of chromatography-mass spectrometry is applied to the quantitative detection of glucose in blood; however, because glucose needs to go through the process of gas phase separation or liquid phase separation, this method is expensive. When the disadvantages, can not achieve high-throughput
Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry has the advantages of accuracy, high throughput, high sensitivity and low sample consumption, and is expected to play an important role in the detection of glucose in blood. However, there are matrix signal interference and The problem of poor signal reproducibility, which caused great difficulties for the quantitative detection of glucose in blood

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  • Method for rapidly, sensitively and quantitatively detecting glucose in serum
  • Method for rapidly, sensitively and quantitatively detecting glucose in serum
  • Method for rapidly, sensitively and quantitatively detecting glucose in serum

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Embodiment 1

[0024] The features and advantages of the present invention can be further understood through the following detailed description in conjunction with the accompanying drawings. The examples provided are only illustrative of the method of the present invention and do not limit the rest of the present disclosure in any way. [Example 1] 2-(4-phenylboronic acid) isoquinolinium bromide labeling combined with black phosphorus assisted laser desorption ionization-time-of-flight mass spectrometry for the detection of glucose in serum

[0025] Preparation of standard curve: Prepare solutions containing glucose standards, the concentrations of which are 0.02, 0.2, 2, 5, 10, 20, 50mmol / L, in which the isotope internal standard [ 2 h 4 ] Glucose was fixed at 5.56mmol / L, and the samples of each concentration were done in parallel three times. After chemically labeling with 10mg / mL 2-(4-phenylboronic acid) isoquinolinium bromide, the samples were classified according to the concentration fr...

Embodiment 2

[0033] [Example 2] 1-(4-phenylboronic acid)pyridinium bromide labeling combined with black phosphorus assisted laser desorption ionization-time-of-flight mass spectrometry for the detection of glucose in serum

[0034] Accurately measure 0.5 μl of serum and place it in a 200 μl centrifuge tube, add 0.5 μl, 2 mg / ml isotope internal standard [ 2 h 4 ] Glucose, ultrasound for 30 seconds. Add 1.5 microliters of acetonitrile, vortex for 30 seconds and centrifuge at 12,000 rpm for 1 minute. Transfer the supernatant to a new 200 μl centrifuge tube, add 0.5 μl, 10 mg / mL 1-(4-phenylboronic acid) pyridinium bromide, and react in a 700W microwave oven on high for 3 minutes. 0.4 μl of the reacted solution was taken for black phosphorus-assisted laser desorption ionization-time-of-flight mass spectrometry analysis. Such as Figure 4 As shown, 3 is the 1-(4-phenylboronic acid) pyridine-labeled product of endogenous glucose detected in serum. For the qualitative determination of glucose...

Embodiment 3

[0036] [Example 3] The method of the present invention is used for the detection of glucose in the serum of clinical samples

[0037] We obtain 54 serum samples clinically, have measured the glucose content in every serum sample with the method of clinical hexokinase; We apply the method of the present invention to measure its glucose content again, and two The results were analyzed by linear regression. Such as Image 6 As shown, although there are differences between the two results, there is a strong correlation, and the linear regression coefficient is 0.9754, indicating that the present invention can be applied clinically to carry out accurate, high-throughput and highly sensitive quantification of glucose in serum detection.

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Abstract

The invention discloses a method for rapidly, sensitively and quantitatively detecting glucose in serum and belongs to the field of biochemical reagent detection. The method comprises the following steps: firstly removing protein in the serum by adopting an organic solvent, carrying out chemical labeling by virtue of a boric acid group and a quaternary ammonium group which are contained in a structure, then adopting black phosphorus-assisted laser desorption ionization-flight time mass spectrometry, and then realizing quantitative detection on the glucose in the serum. The method disclosed by the invention is simple and rapid in operation, and the whole experimental procedure can be completed within 5 minutes; sample consumption amount is low, and endogenous glucose detection can be realized in 0.5 microliter of the serum only; and result is accurate, and large-scale sample determination requirement is met.

Description

technical field [0001] The invention belongs to the field of biochemical reagent detection, and in particular relates to an accurate, rapid and sensitive quantitative detection method for glucose in serum. Background technique [0002] Glucose is a hexose sugar. Most of the energy required for the activities of tissue cells in the human body comes from glucose in the blood. The level of glucose in the blood is an important indicator for the diagnosis of diabetes. Moreover, the continuous increase of glucose levels in the blood can cause a series of complications, such as retinopathy, neurological diseases and cardiovascular diseases. In recent years, the close relationship between glucose content in the blood and cancers such as pancreatic cancer, liver cancer, and colon cancer has also received extensive attention. Therefore, many methods for detecting glucose in blood have been successively reported. [0003] In recent years, glucose sensors based on electrochemical and...

Claims

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Application Information

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IPC IPC(8): G01N27/62
CPCG01N27/628
Inventor 冯钰锜王青余磊丁俊何小梅
Owner WUHAN UNIV
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