Molecular weight analyses of the purified chondroitin and extracted CS were performed as previously described (Restaino et al., 2019 (link)) by a high-performance size-exclusion chromatographic system (Malvern, United Kingdom), equipped with a triple detector array module including a refractive index detector (RI), a four-bridge viscosimeter (VIS), a laser detector (LS) made of a right-angle light scattering (RALS) detector and a low-angle light scattering (LALS) detector, and two gel-permeation columns (TSK-GEL GMPWXL, 7.8 × 30.0 cm, Tosoh Bioscience, Italy). Polyethylene oxide (PEO) was used as a standard for the instrument calibration (22 kDa PolyCAL, Viscotek, Malvern). The average molecular weight, polydispersity index (Mw/Mn), and intrinsic viscosity (IV) were determined by all the detector signals applying the equations reported by the manufacturer (data from Viscotek) and based on the dn/dc values of BC and CS equal to 0.155 and 0.146, respectively.
Polycal
Manufactured by Malvern Panalytical
PolyCAL is a calibration standard developed by Malvern Panalytical for particle size analysis instruments. It provides a traceable size distribution for the calibration and verification of particle size measurement systems.
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Lab products found in correlation
2 protocols using polycal
1
Chondroitin Sulfate Molecular Analysis
2
Absolute Molecular Weight Analysis
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Analysis by SEC/GPC was performed on a ViscotekGPCmax; VE2001 GPC solvent/sample module equipped with triple detector: Refractive Index (RI), Light Scattering (RALS and LALS) and on-line Viscometer. The technique leads to absolute molecular weight as well as data on solution properties. The inert, Mixed Bed ViscoGel I-Series columns: I-MBLMW-3078 and I-MBLMW-3078 (Viscotek) were used to make the measurements. For this purpose, the polymer was dissolved in tetrahydrofuran (THF) at 10 mg/ml at room temperature under soft magnetic shaking for 48 h. After that, 100 μlof each sample was filtered and automatically injected at a flow rate of 0.8 ml/min while the columns and the detector temperature were kept at 30 °C. The determination of the molecular weight distribution was based on the universal calibration technique (UNICAL), which used polystyrene, PS400K and PS65K standards (PolyCAL/Viscotek). Raw data were processed using the OmniSEC GPC/SEC Chromatography System software version 4,5,0,257. Table 1 summarizes the weight average molecular weight (M W ), the polydispersity and the Mark-Houwink exponent, α estimated by this technique.
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