Method for detecting content of CpG ODN in adsorption type vaccine
A vaccine and content technology, applied in the direction of measuring devices, anti-vector-borne diseases, instruments, etc., can solve the problems of peak broadening, tailing or even bifurcation, unsuitable dissociation, etc., to achieve high accuracy and avoid interference Effect
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[0036] (3) Preparation of mobile phase:
Mobile phase A: prepare a 25-100 mmol / L TEAA aqueous solution with a pH of 5.0-9.0.
[0037] Mobile phase B: chromatographic grade acetonitrile.
[0038] Before use, the mobile phase must be vacuum filtered with a 0.45µm mobile phase filter membrane, and sonicated in a water bath for 10-20 minutes to fully degas.
[0039] (4) Processing of CpG ODN compound adjuvant vaccine samples:
See 1. Sample Handling.
[0040] (5) Preparation of CpG ODN standard:
CpG ODN standard stock solution (1 mg / mL): Take 2 bottles of 1 mg CpG ODN standard powder, add 1 mL of purified water to dissolve, and obtain 2 bottles of CpG ODN standard stock solution with a concentration of 1 mg / mL. The above preparation can be prepared in proportion.
[0041] CpG ODN working standard solution (20 μg / mL): Take one bottle of CpG ODN standard stock solution, pipette 200 μL into a 10 mL volumetric flask, and prepare a standard solution with a concentration of 20 μg / ...
Embodiment 1
[0047] experimental method:
[0048] (2) Equipment
[0049] (3) Preparation of mobile phase
[0050] Mobile phase B: Measure 1000 mL of chromatographically pure acetonitrile.
[0051] Before use, the mobile phase must be vacuum filtered with a 0.45µm mobile phase filter membrane, and sonicated in a water bath for 10-20 minutes to fully degas.
[0052] (4) Preparation of CpG ODN standard solution:
[0053] (5) Chromatographic conditions are shown in Table 2.
[0054] Table 2
[0055] Treatment of CpG ODN Compound Adjuvant Novel Coronavirus Inactivated Vaccine Samples: Mix the finished vaccine product and the desorbent in a ratio of 1:1, and place them at room temperature for 1 h; centrifuge the desorbed samples at 15,000 rpm / min for 15 min at room temperature. Take the supernatant solution.
[0056] The samples treated according to step (6) were analyzed by the above HPLC detection method. The control samples were samples without desorbent treatment. The experimental resu...
Embodiment 2
In this example, 6 batches of CpG ODN compound adjuvant vaccines were detected by this method, specifically the finished products of CpG ODN adjuvant novel coronavirus virus inactivated vaccines.
[0058] The experimental method is the same as in Example 1.
[0059] Edit the sequence table, apply the established run method, and equilibrate the system with mobile phase until the baseline is stable (takes approximately 1 hour). Execute the sequence and, after the injection sequence is complete, process the data from each injection to ensure that the target analyte peaks are correctly integrated.
[0060] The experimental results are shown in Table 4:
Table 4
The chromatograms of 6 batches of finished products are as follows Figures 2A-2F shown.
[0061] The linear curve of CpG ODN standard is shown in the figure image 3 shown.
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