Valproic acid quantification in human serum by LC-MS/MS: method development and comparison with immunoassay

Adriano Ronchi Spillere; Laura Alencastro de  Azevedo; Juliana Severo Almeida; Sarah Carobini Werner de Souza Eller Franco de  Oliveira; Tiago Franco de Oliveira; Bruno Pereira dos Santos; Simone Martins de  Castro

doi:10.22456/dar.v10i1.154530

Authors

Adriano Ronchi Spillere Faculdade de Farmácia - Universidade Federal do Rio Grande do Sul https://orcid.org/0009-0000-4450-1104
Laura Alencastro de Azevedo Faculdade de Farmácia, Laboratório de Análises Clínicas e Toxicológicas - Universidade Federal do Rio Grande do Sul https://orcid.org/0000-0002-4982-8832
Juliana Severo Almeida Faculdade de Farmácia, Laboratório de Análises Clínicas e Toxicológicas - Universidade Federal do Rio Grande do Sul https://orcid.org/0009-0001-6924-0788
Sarah Carobini Werner de Souza Eller Franco de Oliveira Faculdade de Farmácia, Laboratório de Análises Clínicas e Toxicológicas - Universidade Federal do Rio Grande do Sul; Programa de Pós-graduação em Ciências da Saúde - Universidade Federal de Ciências da Saúde de Porto Alegre https://orcid.org/0000-0003-2200-6959
Tiago Franco de Oliveira Faculdade de Farmácia, Laboratório de Análises Clínicas e Toxicológicas - Universidade Federal do Rio Grande do Sul; Programa de Pós-graduação em Ciências da Saúde - Universidade Federal de Ciências da Saúde de Porto Alegre https://orcid.org/0000-0003-2594-8415
Bruno Pereira dos Santos Faculdade de Farmácia, Laboratório de Análises Clínicas e Toxicológicas - Universidade Federal do Rio Grande do Sul; Programa de Pós-graduação em Ciências da Saúde - Universidade Federal de Ciências da Saúde de Porto Alegre https://orcid.org/0000-0002-7624-5966
Simone Martins de Castro Faculdade de Farmácia, Laboratório de Análises Clínicas e Toxicológicas - Universidade Federal do Rio Grande do Sul https://orcid.org/0000-0001-6707-9698

DOI:

https://doi.org/10.22456/dar.v10i1.154530

Keywords:

valproic acid, immunoassay, liquid chromatography-mass spectrometry, therapeutic drug monitoring

Abstract

Valproic acid (VPA) is widely used for epilepsy management, however VPA serum concentration must be monitored because its narrow therapeutic range. Some methods are used to determine the plasma or serum VPA concentration, among them immunoassay-based tests are the most used. Although immunoassays may present cross-responses with other drugs or AE metabolites, increasing the uncertainty of the method. For this reason, a liquid chromatography-mass spectrometry in tandem (LC-MS/MS) method was developed, validated and compared with particle-enhanced turbidimetric inhibition immunoassay (PETINIA). For method optimization, the protein precipitation solvent was methanol and isopropanol (3:1) and a biphenyl column (50 x 3.0 mm, 2.7 μm) was used. The method showed selectivity and specificity, and it was linear from 25 to 450 µg/mL, presenting variation coefficient lower than 8,4% and inaccuracy within 10,9%. The matrix effect was 103.9%. The VPA quantification by both methods had good correlation and the VPA assay by LC-MS/MS was 6,85% higher when compared to the immunoassay, but it still gives the same clinical stratification. The validation was successful, with easy sample preparation and appropriate selectivity, applicable for VPA therapeutic drug monitoring.

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