Optimization and Validation of the HPAEC-PAD Method for Quantifying Vi Polysaccharide in the Typhoid Vaccine
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Abstract
Background: The quantitative assessment of Vi polysaccharide in the typhoid vaccine plays a crucial role in ensuring vaccine quality control. It verifies that the specific antigen in the typhoid vaccine corresponds to the information in the registration dossier, thereby guaranteeing the quality, efficacy, and safety of the vaccine. HPAEC-PAD is known for its high accuracy and precision, establishing it as the gold standard method recommended by WHO for the analysis of carbohydrates, oligosaccharides, and polysaccharides.
Objective: To optimise and validate the HPAEC-PAD method for measuring the amount of Vi polysaccharide in the typhoid vaccine, intended for use as a standard method in the laboratory.
Methods: The HPAEC-PAD method was optimized and validated across multiple parameters, such as linearity and range, precision, accuracy, specificity, limit of detection, limit of quantification, and robustness.
Results: The validation of the HPAEC-PAD method showed a linear standard curve in the concentration range of 5 – 40 µg/ml, with a linear regression equation of y = 28.33x + 0.7000 and a correlation coefficient of 0.9999. The precision test conducted on the same day and on different days showed relative standard deviation percentages of 2.25 and 4.44, respectively. The accuracy test demonstrated that HPAEC-PAD can be used to measure the standard substance in the typhoid vaccine, with a percentage of recovery between 92.90 and 115.22. The limits of detection and quantitation of HPAEC-PAD were 2.39 and 10.00 µg/mL, respectively. The specificity test of the HPAEC-PAD method confirmed the absence of interference from other substances and its ability to isolate the target antigen. Additionally, the HPAEC-PAD method demonstrated robustness even with changes in analysts, and all validation method parameters met the acceptance criteria.
Conclusions: The validation of the HPAEC-PAD method for quantifying the amount of Vi polysaccharide in the typhoid vaccine demonstrates that the method met the acceptance criteria. Therefore, the HPAEC-PAD method is suitable for use as a standard laboratory procedure, thus ensuring that consumers receive high-quality vaccines for disease prevention.
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References
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