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
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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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