alpha,beta-Poly(asparthylhydrazide) (PAHy) is a new synthetic polymer that exhibits interesting properties and is a candidate for biomedical applications. In this article the characterization of PAHy polymer by multi-angle laser light scattering (MALS) and single-capillary viscometer (SCV) detectors on-line to a size exclusion chromatography (SEC) system is reported. The SEC-MALS-SCV system furnishes exhaustive and consistent characterization of the PAHy polymer. Further, it is possible to characterize the PAHy polymer through conventional SEC and universal calibration. The universal calibration method gives intrinsic viscosity and dispersity very close to those measured by the absolute detectors; instead the weight-average molar mass is approximately 8% lower. This finding means that the chromatographic separation of PAHy polymer is fundamentally based on its hydrodynamic volume. By the SEC-MALS-SCV system the constants of the Mark-Houwink-Sakurada equation were also estimated.