Method for rapidly detecting dextran sulfate immobilized amount in blood lipid adsorbent

A dextran sulfate and adsorbent technology, applied in the biological field, can solve the problems of complicated operation, complicated calculation steps, and high reagent concentration, and achieve the effects of good repeatability of results, reliable experimental results, and small amount of reagents

Active Publication Date: 2018-11-02
WUHAN RUIFA MEDICAL DEVICES CO LTD
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the detection methods for dextran sulfate include hydrochloric acid hydrolysis-barium sulfate gravimetric method, resonance Rayleigh scattering method, liquid chromatography and polytoluidine blue molecular probe method. These methods have higher requirements for instruments or reagents. And it can only detect aqueous samples, but cannot detect the content of dextran sulfate immobilized on the microspheres
Among them, the principle of using polytoluidine blue as a molecular probe to identify dextran sulfate is similar to this method, but a layer of polytoluidine blue film needs to be electropolymerized on a glassy carbon electrode, which is complicated and time-consuming, and the method Also only suitable for aqueous samples
[0006] Patent CN1697665A relates to a method for detecting the fixed amount of dextran sulfate. Through the change of toluidine blue content before and after adsorption, the amount of toluidine blue reduction is calculated, and then the fixed amount of dextran sulfate is obtained through a certain conversion relationship. The calculation steps of this method are complicated, and the concentration of the toluidine blue reagent used is large. When detecting the absorbance at 630nm, multiple dilutions are required, and the error in the measurement is multiplied. In the actual experimental operation, the absorbance of the toluidine blue solution shows a downward trend, resulting in There is a large error
Therefore, the calculation of this method is complicated, the results have large errors and the amount of reagents is large

Method used

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  • Method for rapidly detecting dextran sulfate immobilized amount in blood lipid adsorbent
  • Method for rapidly detecting dextran sulfate immobilized amount in blood lipid adsorbent
  • Method for rapidly detecting dextran sulfate immobilized amount in blood lipid adsorbent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The establishment of embodiment 1 standard curve and detection range

[0046] Weigh 0.2g of dextran sulfate powder, accurately weigh it, dissolve it in 50ml of pure water, use it as a stock solution (4mg / ml), and store it in a refrigerator at 4°C.

[0047] Drawing of the standard curve: dextran sulfate solution with a concentration of 4mg / ml was used as the mother solution, and diluted with pure water in different times. , 35, 40 μg / ml, adopt the inventive method to carry out dextran sulfate content detection (acid reagent adopts hydrochloric acid, final concentration is 0.002mol / L), when concentration >=20 μ g / ml, reaction tends towards saturation, as figure 1 shown.

[0048] Within the concentration range of 0-20μg / ml, set a set of standard solution again, the concentration is 0, 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20μg / ml, the standard solution within this range The solution concentration and absorbance have a good linear relationship, and the standard curve R 2 up...

Embodiment 2

[0054] Embodiment 2 matrix interference test

[0055] The choice of substrate in the reaction system has a great influence on the accuracy of the experimental method. PBS buffer is the most widely used buffer in biochemical research. It has the osmotic pressure of physiological saline and has a stable pH value for the prepared solution. Therefore, PBS buffer was selected as the matrix to be investigated in the experiment. When PBS buffer was added to the reaction system instead of pure water, there was no obvious color change in the reaction, and there was almost no difference in the absorbance measured at 630nm for different concentrations of standard substances, indicating that the solution matrix inhibited the reaction.

[0056] At the same time, refer to the relevant literature of the toluidine blue photometric method. When various ions are used as coexisting substances in the reaction system, they will affect the detection results to varying degrees. In order to eliminate t...

Embodiment 3

[0057] Embodiment 3 stability test

[0058] In order to ensure the accuracy of the numerical value, the stability of the absorbance value of the sample needs to be tested, and the content of dextran sulfate is detected by a common method (the difference from the method of the present invention is that in step (2), no acid reagent is added, and the other steps are the same) , standing still for 10min, 30min, and 50min, samples were taken for detection, and the absorbance value showed an obvious downward trend. The standard solution with a concentration of dextran sulfate of 0 μg / ml was used as the test sample, and the measurement results are shown in Table 2.

[0059] Table 2 Untreated sample stability determination result

[0060] Standing time (min)

[0061] Adopt the method of the present invention to carry out dextran sulfate content detection (acid reagent adopts hydrochloric acid or hydrochloric acid and gallic acid, in step (2), add the hydrochloric acid 100 μ ...

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Abstract

The invention discloses a method for rapidly detecting a dextran sulfate immobilized amount in a blood lipid adsorbent. According to the method, an acid reagent is added to stabilize a light absorption value after reaction step completion of toluidine blue and dextran sulfate, so that the experimental result is reliable. Moreover, the reagent usage amount in the detection process is small, the calculation is simple, the result repeatability is excellent, and the accuracy is high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for rapidly detecting the fixed amount of dextran sulfate in a blood lipid adsorbent. Background technique [0002] Many epidemiological, genetic and clinical studies have confirmed that the level of low-density lipoprotein cholesterol (LDL-C) in serum is positively correlated with the incidence of atherosclerosis (AS) and coronary heart disease (CHD), usually with high LDL-C C level is the primary pathogenic factor of CHD. The American National Cholesterol Education Program (NCEP) adult treatment professional group stipulates that LDL-C level is used as the basis for the classification and treatment of hyperlipidemia, as well as the treatment goals that need to be achieved. [0003] Dextran sulfate is a polyanionic derivative of dextran, formed by the esterification of dextran with chlorosulfonic acid. Because its structure and charge are similar to the LDL-C receptor, wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 王业富张媛张磊胡定邦温佳文何改粉
Owner WUHAN RUIFA MEDICAL DEVICES CO LTD
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