Acid hydrolysis conditions for quantification of meningococcal X polysaccharide in a pentavalent vaccine using HPAEC-PAD/ESI-MS

A selective and sensitive method was evaluated for quantitation of meningococcal X (Men X) polysaccharide in pentavalent meningococcal A, C, W, Y and X conjugate vaccine using different acid hydrolysis conditions like HCl, TFA, HF, HF-TFA, and HF–HCl. High-performance anion exchange chromatography w...

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Bibliographic Details
Published in:Analytical biochemistry 2023-12, Vol.683, p.115363-115363, Article 115363
Main Authors: Ghosh, Saurav, Pawar, Rakesh, Kangralkar, Vivek, Mallya, Asha D., Dhere, Rajeev M., Bolgiano, Barbara, Ravenscroft, Neil
Format: Article
Language:English
Subjects:
Acid Hydrolysis
Carbohydrate-based vaccine
Chromatography, Ion Exchange - methods
Electrospray mass spectroscopy
Glucosamine
HPAEC-PAD
Humans
Hydrolysis
Meningococcal A, C, W, Y, and X polysaccharides
Meningococcal Vaccines - analysis
Meningococcal Vaccines - chemistry
Neisseria meningitidis
Polysaccharides, Bacterial - analysis
Polysaccharides, Bacterial - chemistry
Tandem Mass Spectrometry
Vaccines, Combined
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Summary:A selective and sensitive method was evaluated for quantitation of meningococcal X (Men X) polysaccharide in pentavalent meningococcal A, C, W, Y and X conjugate vaccine using different acid hydrolysis conditions like HCl, TFA, HF, HF-TFA, and HF–HCl. High-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) using CarboPac PA10 column was used to identify the hydrolyzed products based on retention time and its comparison with monosaccharide standards. Complete release of glucosamine (GlcN) from Men X in monovalent bulk and pentavalent vaccine samples was achieved using HF hydrolysis at 80 °C for 2 h. The Men X HF-hydrolyzed polysaccharide to glucosamine along with the reference standard was identified using collision-induced dissociation (CID) electrospray mass spectroscopy and the MS/MS fragments of m/z 162, m/z 144 and m/z 84. Meningococcal polysaccharide concentration was determined with a correlation coefficient r2 >0.99 using polysaccharide reference standard. The serogroups A, W, and Y were converted to their monosaccharides units and quantified using this method however, milder acid hydrolysis 0.1 M HCl 80 °C 2 h for release of sialic acid for Men C polysaccharide was found to be more suitable. These methods will provide necessary tools and prove to be beneficial to laboratories developing new saccharide-based vaccine combinations. [Display omitted] •Meningococcal polysaccharide hydrolysis performed using various acids to convert into monosaccharide units.•Characterization of the monosaccharide glucosamine post Men X after HF hydrolysis using mass spectrophotometric analysis.•HF hydrolysis at 80 °C for 2 h method for quantitative determination of pentavalent meningococcal conjugate vaccine.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2023.115363