Validated LC-MS/MS quantification of kynurenine pathway metabolites and pathway-balance ratios in brain tissue and microglial culture supernatant: application to neonatal stroke

An LC-MS/MS assay quantifying eight kynurenine pathway metabolites and four balance ratios in neonatal rat brain tissue and BV2 microglial supernatant shows LOQ 6.53–34.17 nM and R2 > 0.99.

• Why it matters: KP metabolite imbalance is linked to perinatal brain injury, but validated measurement methods across brain tissue, glial media, and human biofluids are limited, hindering direct comparisons.
• What they did: The workflow used protein precipitation plus Enhanced Matrix Removal-Lipid clean-up, then met CLSI acceptance criteria in brain homogenate and cell supernatant, and was applied to neonatal stroke hemispheres and LPS/cytokine-stimulated BV2 cells.
• The result: In the stroke hemisphere, KYN/TRYP and QA/KYNA were higher and KYNA/KYN lower at 1, 7, and 14 days (p<0.001), with BV2 showing the same four-ratio shifts (p<0.01), supporting KYN/TRYP and QA/KYNA as candidate targets for neonatal biofluid studies.

AI summary of the abstract · may contain errors · not medical advice · read the paper.