Development of an Accurate Double Isotopic Standard LC-MS/MS Method for Hyaluronic Acid Quantification in Biological Matrices.

ABSTRACT: Hyaluronic acid (HA) plays key roles in tissue hydration, repair, and cellular signaling. Its quantification in biological matrices is crucial but challenging due to its endogenous nature and poor mass spectrometric detectability. We developed a robust method based on enzymatic hydrolysis, dual ¹³C-labeled internal standards, and the standard addition method combined with LC-MS/MS analysis. Samples from bovine vitreous humor and human synovial fluid were depolymerized with recombinant hyaluronidase to generate Δ4-mer oligomers, quantified using 100%- and 50%-¹³C-labeled HA as internal standards to correct the variabilities of the enzymatic digestion and MS detector. The standard addition method was used to control the matrix effects. The method showed excel

SUBMITTER: Manzi S 

PROVIDER: S-EPMC12937047 | biostudies-literature | 2026 Feb

REPOSITORIES: biostudies-literature