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Size exclusion chromatography with UV detection (SEC‐UV) is a precise method for quantifying polyacrylamides in water. However, its accuracy can be compromised in complex matrices due to chemical degradation of the polymer. This study focuses on the impact of hydrolysis on quantifying sulfonated polyacrylamide copolymers (PAM‐AMPS) via SEC‐UV, analyzing key influencing factors. Using controlled hydrolysis, hydrolyzed copolymer (HPAM‐AMPS) standards with varying hydrolysis degree (HD) (HD = 7%–33%) were prepared without significant changes in polymer chain size. SEC‐UV analysis revealed that quantification accuracy is highly dependent on hydrolysis extent. Errors are minimal for HD up to 10%, but significantly higher for HDs of 22%–33%. Furthermore, this effect is modulated by polymer concentration. Our findings indicate that there are two compensating factors for the decrease in molar absorptivity due to amide‐to‐carboxylate substitution: the sulfonate group acting as an additional chromophore, and a redshift in absorption spectra due to hydrogen bonding at higher concentrations. This interplay between HD and concentration in this type of sulfonated polyacrylamide can help reduce quantification errors in complex aqueous samples prone to chemical degradation, like those found in environmental and subsurface applications. [ABSTRACT FROM AUTHOR]