Hepatitis B virus core antigen content detection method

A core antigen, hepatitis B virus technology, applied in the medical and health field, can solve the problems of detection and hepatitis B virus core antigen, and achieve the effects of simple operation, shortened detection time, and simplified quantitative analysis

Inactive Publication Date: 2016-12-21
林海燕
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the technical defects of the prior art, and provides a method for detecting the content of hepatitis B virus core antigen, so as to solve the technical problem that the hepatitis B virus core antigen is difficult to be directly detected from serum in the prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Chromatographic conditions

[0023] Choose Shimadzu C8 column (4.5*130mm, 4μm), the mobile phase is a mixture of 0.12mol / L ammonium acetate solution (pH8.5) and acetonitrile (40:60, V / V), the flow rate is 0.7ml / min, the column temperature The drift tube temperature of the evaporative light scattering detector is 55°C, and the gas flow rate is 22psi. The injection volume of the autosampler was 17 μL.

[0024] (2) Drawing of standard curve

[0025] Take 50 mg of HBcAg standard substance, dissolve it with chromatographic grade acetonitrile, dilute to 50 mL, and dilute in turn to form standard solutions with HBcAg content of 5, 10, 20, 50, 100, 200, and 500 μg / ml, and add them to step (1) respectively. 20 μl in the chromatographic column of the high-performance liquid chromatograph of running state, calculate the peak area under the same retention time respectively, take concentration logarithm log10 (C) as abscissa, take peak area logarithm log10 (A) as ordinate, dra...

Embodiment 2

[0036] (1) Chromatographic conditions

[0037] Select the Agilent C8 column (the length of the C8 chromatographic column is 150mm, the inner diameter is 4.3mm, and the average particle diameter of the filler is 4μm), and the mobile phase is 0.12mol / L ammonium acetate solution (pH8.5) and acetonitrile (50:50, V / V) mixture, the flow rate is 0.5ml / min, the column temperature is 25°C, the drift tube temperature of the evaporative light scattering detector is 55°C, and the gas flow rate is 22psi. The injection volume of the autosampler was 17 μL.

[0038] (2) Drawing of standard curve

[0039] Take 50 mg of HBcAg standard substance, dissolve it with chromatographic grade acetonitrile, dilute to 50 mL, and dilute in turn to form standard solutions with HBcAg content of 5, 10, 20, 50, 100, 200, and 500 μg / ml, and add them to step (1) respectively. 20 μl in the chromatographic column of the high-performance liquid chromatograph of running state, calculate the peak area under the sa...

Embodiment 3

[0045] A detection method for hepatitis B virus core antigen content, the method belongs to the HPLC method, the method utilizes a C8 chromatographic column to separate, and utilizes an ELSD detector to detect; its chromatographic conditions are as follows: mobile phase consists of pH8.5, concentration 0.12mol / L Ammonium acetate solution and acetonitrile, wherein acetonitrile accounts for 70% (v / v) of the total mobile phase; flow rate is 0.9mL / min; column temperature is 35°C.

[0046] On the basis of the above technical solutions, the following conditions are met:

[0047] The temperature of the drift tube of the ELSD detector was 55° C., the gas flow rate was 22 psi, and the injection volume was 17 μL.

[0048] The length of the C8 chromatographic column is 150mm.

[0049] After detection, the HBV core antigen was quantified by the internal standard method.

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Abstract

The present invention provides a hepatitis B virus core antigen content detection method. According to the technical scheme, hepatitis B virus core antigen is detected by using a HPLC method, a detector and chromatographic conditions are optimized, the detection precision is remarkably improved, and the trace HBcAg can be detected after the serum is directly taken out without the addition of a shell opening agent; the mobile phase and the solvent of an ELSD detector can evaporate completely during a detection process, such that the obtained chromatogram does not have the solvent peak, and the gradient elution does not have the reflected light parallax effect and cannot generate the baseline drift so as to significantly simplify the subsequent quantitative analysis; the retention time of the C8 column is short, and the separation effect is good, such that the detection time is shortened; and the detection conditions are mild, such that the denaturation deactivation of proteins, nucleic acids and other components cannot be caused so as to effectively ensure the accuracy of the detection. According to the present invention, the excellent technical effect is achieved by using the innovative technical improvement while the advantages of high sensitivity and easy operation are provided, and the accurate quantitation can be achieved by using the internal standard method after the detection.

Description

technical field [0001] The invention relates to the field of medical and health technologies, in particular to a method for detecting the content of hepatitis B virus core antigen. Background technique [0002] Hepatitis B is a chronic infectious disease caused by hepatitis B virus (HBV) infection. The initial symptoms are hidden, and the course of the disease is longer. my country is a country with a high incidence of hepatitis B, and the carrier rate of hepatitis B surface antigen accounts for about 7-8% of the total population. Therefore, technologies for the prevention, treatment, and detection of hepatitis B are very important. [0003] At present, the clinical detection of HBV is mainly through five indicators of surface antigen (HBsAg), surface antibody (anti-HBs), e antigen (HBeAg), e antibody (anti-HBe) and core antibody (anti-HBc). Hepatitis B two-and-a-half) for evaluation. The existence of the above five substances has a relatively stable corresponding relation...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 林海燕
Owner 林海燕
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