New application note from Wyatt demonstrates the accurate and precise determination of molecular weight and concentration of sodium hyaluronate
Wyatt Technology has published a new free-of-charge application note to demonstrate the use of Sec-Mals for the determination of molecular weight (MW) and concentration of sodium hyaluronate (HA, 1.6-2.9MDa) in mobile phase solution and mixed with gelatine.
Wyatt's powerful Sec-Mals instruments were found to ascertain molecular weight and concentration of HA at very high mass recovery.
HA, a naturally-occurring biopolymer in all mammalian systems, is of huge commercial interest for medical, pharmacological, ophthalmic, and cosmetic applications.
Using its Optilab Rex differential refractometer and Dawn Heleos, Wyatt has overcome consistent problems experienced using other techniques to accurately calculate HA's characteristics.
HA has been studied considerably using methods such as size-exclusion chromatography (SEC) and single-capillary viscometry (SCV).
Wyatt Technology has allowed scientists to consistently investigate the material characteristics of HA by combining its innovative instrumentation with SEC.
For this particular application, a Dionex system was connected to a Wyatt Optilab Rex differential refractometer and a Dawn Heleos.
TSK-GEL G6000PWXL and G5000PWXL (Tosoh Bioscience) columns were utilised and the mobile phase was 0.20M PBS (ph 7).
Tests for specificity revealed that it was possible to detect HA with MWs of 1.6, 2.4, 2.6 and 2.9MDa.
Surprisingly a mass recovery of 99.7% for HA mixed with gelatin was achieved.
The MW was determined to be 86.3% of the expected MWHA.
The sample was treated with a protease solution during heating at 40C.
Samples with pure gelatin and protease showed no peaks in the HA area.
Heating 2.6 MDa HA solutions (0.15mg/ml) to 80C yielded degradation products with MW of 1.9MDa and mass recovery of 97%.
Results of the application establish the employment of Wyatt's instruments to guarantee exceptional, rapid, precise analysis.
Results determined the absolute HA molecular weight and concentration of HA when mixed with gelatine at a very high mass recovery (99.7%) and MWobs/MWHA in the same range as for pure HA in solutions.
