The Eclipse AF4 combines with a DAWN HELEOS II
MALS detector and Optilab T-rEX dRI detector to
solve the most difficult separation, fractionation and sizing tasks in the macromolecular and colloidal arenas including polymers, biotechnology,
pharmaceuticals, nanotechnology and environmental analysis.
The advantage of AF4 over traditional SEC column chromatography is the ability to separate both soluble and colloidal components over a wide size
range as well as sensitive and "sticky" samples, since there is no stationary phase. AF4 even overcomes the non-ideal behavior exhibited by large,
branched polymers that elute abnormally on GPC columns. The Eclipse provides resolution similar to that of analytical centrifugation (AUC) but
surpasses even this benchmark tool when one considers its flexibility, speed, and ease of use.
Common solution-based methods for determining the size and size distributions of nanoparticles, such as batch dynamic light scattering (DLS) and
nanoparticle tracking analysis (NTA), cannot offer the resolution, statistical confidence and absolute accuracy of a true separation-based technique
that incorporates MALS and DLS downstream detection.
FFF-MALS and FFF-DLS performed with an Eclipse AF4,
DAWN HELEOS II and an embedded WyattQELS DLS
detection module offer excellent resolution and accuracy in analysis of size distributions. Particle counts and sizes of viruses and virus-like
particles estimated by FFF-MALS match closely TEM-based counts.
Key benefits of FFF with a Wyatt Eclipse:
Robust separation of macromolecules and nanoparticles from 1 nm - 1000 nm with excellent resolution
No stationary phase - open channel separation with minimal shear and surface interactions
Versatile choice of separation channels, permitting sample loads from µL to mL, including semi-preparative FFF
Optimize separation methods to enhance resolution or size range simply by adjusting flow ratios, even programmatically during a run
Fractions can be collected and used for off-line analysis (with electron microscopy, MS, ICP-MS, ELISA, etc.)
... and of the Eclipse AF4:
Typical analysis times 10 - 30 minutes
Utilize the fritless column for aggregation-prone samples
Make use of the highest quality HPLC pumps and autosamplers from Agilent, Shimadzu, Thermo, or Waters
Integrate with your favorite HPLC software from Agilent, Dionex or Shimadzu
Couple to ICP-MS for elemental analysis of nanoparticles per size bin