The ultimate separation system for proteins, polymers and nanoparticles.
Flow Field-Flow Fractionation is a uniquely versatile means of size-based separations, capable of fractionating and characterizing proteins and aggregates, liposomes, emulsions, viral particles, polysaccharides, nanoparticles, polymer latex particles, colloidal soil suspensions, chemical mechanical polishing slurries, and many more - with no stationary phase and almost no shear.
The Eclipse™ DualTec™ system offers both Hollow-Fiber Flow-FFF (HF5) and Asymmetric Flow-FFF (AF4) techniques. Both may be integrated into one instrument and coupled seamlessly to the most advanced light scattering detector for determination of molar mass and nanoparticle size, Wyatt's DAWN® instrument.
Macromolecules: Complement SEC-MALS
Eclipse FFF systems add to, and complements, size exclusion chromatography (SEC) for macromolecular separations. Separation occurs without shear or adverse, non-ideal column interactions. The Eclipse DualTec allows you to take full advantage of your favorite conventional SEC and AF4/HF5 separations in one convenient package.
The DualTec lets you use the same HPLC pump and autosampler, and the same MALS and DLS light scattering detectors, to characterize macromolecules for molecular weight, size and conformation by SEC or FFF. Switch between the two separation techniques at the click of a mouse.
Nanoparticles: High-resolution characterization
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 DualTec, DAWN 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
- Typical analysis times 5 to 30 minutes
- No stationary phase - open channel separation with minimal shear and surface interactions
- Versatile choice of separation channels, permitting sample loads from nL 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 DualTec
- Choose between any two of SEC, AF4 or HF5 mode separations with a single HPLC and detector set
- 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
Innovation, innovation, innovation
The innovative Eclipse DualTec makes available two different lines of separation in a single instrument configuration. Each line draws sample from the same autosampler, propelled by the same pump, and characterized by the same detectors. Each separation line can be configured for SEC, AF4 or HF5, independently, so you can:
- Compare and select the most appropriate technique
- Develop a method with tiny sample volumes by HF5, then transfer to AF4 to increase information content*
- Attach two HF5 channels to double the time between replacing cartridges
- Innovate your research with new analytical capabilities
*Method transfer is completed without additional experimental work by applying the SCOUT DPS™ software, a patented procedure to predict and optimize your separation based on the flow rates and channel geometries. SCOUT DPS is sold separately.
Asymmetric-Flow Field-Flow Fractionation (AF4)
AF4, the most versatile and common type of FFF, employs a flat-channel configuration (see FFF-MALS and FFF Theory for more details, or view FFF-MALS webinars). Flat separation channels are available in a variety of sizes to optimize sample loading (which impacts the detection of trace quantities of aggregates, fragments and other inabundant species) against run time and solvent consumption. Channel sizes even run to the semi-preparative scale which can purify milligram quantities of protein. Channels are available with different wetted materials, from the basic PEEK to all-metal (for organic solvents) and metal-free.
AF4 channels are available with two standard membrane materials, regenerated cellulose and polyether sulfone. If these materials do not suit your solvents you can even prepare and install your own membranes, using the convenient cut-out template we provide. Membranes typically last through dozens if not hundreds of injections, then are disposed of and replaced with another inexpensive membrane at the cost of a few dollars and the effort of a few minutes of cleaning the separation chamber. Compared to the cost of a new SEC channel, FFF supplies are insignificant!
The advantages of Hollow-Fiber FFF (HF5)
HF5 separation utilizes a hollow-fiber cartridge made of polyether sulfone. The principle of operation is very similar to AF4 but now the channel is 'rolled' in a tube format instead of flat.
HF5 offers separations with very little volume, high sensitivity, and short analysis time, thanks to the very small volume of the cartridge. You could say that HF5 is the analog of UHPLC, except - like AF4 - there is no stationary phase and no shear.
HF5 is preferred over AF4 for method development, because it takes minimal sample, time and solvent consumption. The Eclipse HF5 channel cartridge typically serves for more than a hundred injections, then may be replaced by a fresh cartridge within seconds. Downtime is minimized, and there is no membrane to install or cleaning step required.
Sensitivity is a primary benefit of HF5, which has the highest sensitivity of all Flow-FFF variants. The reason is the low channel volume - typically below 100 µL - and high crossflow density. Moreover, HF5 has the ideal channel geometry with a completely linear flow path and no depletion wall which supports higher efficiency.
Increase in productivity
HF5 runs on the hollow-fiber are fast and need very little sample amount. The Eclipse DualTec offers disposable HF5 channel cartridges which can be replaced in seconds at minimal costs. After method development the separation can be transferred for scale-up to AF4 channels which allow higher sample load.
Which to choose?
Your choice of solvents may limit your system to AF4 operation; conversely, for aqueous systems sample availability may constrain you to working with HF5. With an Eclipse DualTec, you don't have to choose! You can install both AF4 and HF5, or just one or the other with SEC, and reconfigure as your needs change.
The Eclipse is the world's most advanced AF4 system
- Fully automated, software controlled system with only one pump to generate detector flow, crossflow and injection flow. This innovative concept to generate all three flow streams from one main flow has been a breakthrough for the Flow-FFF method!
- Three DualTec versions: for aqueous solvents (biocompatible), for organic solvents and in a metal-free version
- Proprietary pressure sensors without dead-volume monitor channel and cross-membrane pressures
- The Eclipse DualTec is certified by TÜV Rheinland
Eclipse Separation Channel Designs
The Eclipse DualTec offers the most versatile separation channel technology. There are four different channel types (four are compatible with the DualTec, while the fritless channel requires the Eclipse AF4) plus the hollow-fiber-cartridge available to accommodate different requirements for sample load, analysis time, and resolution. Consumables include a wide choice of spacers and pre-cut membranes for all channel types.
All channels have a metal-free flow path, making them inert even in harsh conditions. All channels are easy to maintain - replacing a membrane or cleaning the channel parts is straightforward, with just a few screws to tighten during the reassembly process.
- Short Channel (SC 145 mm length)
- Long Channel (LC, 275 mm length)
- Hollow-Fiber FFF Channel (HF5)
Integration with Wyatt Technology instruments and market-leading HPLC packages
- The Eclipse separation system is designed to integrate with Wyatt Technology's range of unique light scattering and refractive index detectors, ensuring highest sensitivity and optimum baseline quality.
- The Eclipse uses either Agilent, Shimadzu, or Dionex chromatography components. Keep your current HPLC software platform and run the Eclipse with the familiar interface you already know.
- A single pump generates detector flow, crossflow, and injector flow, achieved by splitting the main flow delivered through the HPLC pump. Using only one pump results in a much improved signal-to-noise ratio because pump pulsations are reduced and balanced, and operation is much simpler.
DAWN - The most sensitive MALS detector available, anywhere. Incorporates detectors at 18 angles to determine molar masses from 200 Da to 1 GDa and radii from 10 – 500 nm. Heated/cooled option -15°C to +150°C.
The DAWN offers advanced options such as fluorescence or polarization filters and a 785 nm laser.
miniDAWN - Second only to the DAWN in sensitivity. Incorporates detectors at 3 angles to determine molar masses from 200 Da to 20 MDa and radii from 10 to 50 nm. Ambient only.
Optilab - A unique on-line differential refractometer for measuring concentration of any macromolecule, regardless of chromophores.
WyattQELS - A dynamic light scattering (DLS) module which integrates inside the DAWN or miniDAWN to provide simultaneous DLS measurements in the same scattering volume. The optical fiber replaces any one MALS detection angle.
Complete FFF-MALS systems
- The Eclipse separation system integrates the Eclipse chassis and channels (on the right) with high-quality HPLC components for liquid chromatography (on the left).
- The Eclipse Flow-FFF allows you to choose from a wide range of pumps, autosamplers, detectors and fraction collectors from the leading HPLC manufacturers Agilent, Dionex, and Shimadzu.
- Only one pump is needed to generate detector flow, crossflow, and injector flow by splitting the main flow delivered through the HPLC pump. Using only one pump results in a much improved signal-to-noise ratio because pump pulsations are reduced and operation is much easier.
- Existing HPLC instruments can also be integrated easily with a new Eclipse system.
- The Eclipse control is integrated in Agilent ChemStation, Agilent OpenLAB, Thermo (Dionex) Chromeleon, or Shimadzu LCsolution software allowing access to the extensive base of hardware which is supported by the HPLC software packages.
- The Eclipse DualTec separation system is designed to integrate with Wyatt Technology's complete range of unique multi-angle light scattering (MALS), dynamic light scattering (DLS), refractive index (RI) and viscometry (VI) detectors, ensuring highest sensitivity and optimum baseline quality.
If you would like to add FFF capability to your existing GPC/SEC column chromatography system, please Contact Us We can offer you an individual solution designed for your specific application and system requirements.
|Standard mode||1 nm to 1000 nm, depending on specific sample properties and separation channels|
|Steric mode||Approximately 1 µm to 10 µm, depending on specific sample properties and separation channels|
|Max. total flow rate||10 mL/min (crossflow plus detector flow rate)|
|Max. channel pressure||30 bar, software overpressure shut-down|
|Spacer thickness||Set of spacers with 250, 350 and 490 µm|
Polyether Sulfone: 1, 3, 5, 10, and 30 kDa cut off
Cellulose Triacetate: 10 kDa cut off
Regenerated cellulose: 5, 10, and 30 kDa cut off
Short SC: 153 mm e.l.|
Long LC: 246 mm e.l.
Disposable, ready-to-use hollow-fiber cartridges:
400 µm radius
PES membrane 10 kDa cutoff
channel volume 90 µL
Two zero-dead-volume pressure sensors with range 0 to 30 bars monitor pressure across the membrane and in the channel;
Coriolis-effect flow regulators are insensitive to solvent type and temperature;
Rated for aqueous solvents and alcohols
Guaranteed corrosion resistant from pH 2 to 12
Wetted materials: PEEK, stainless steel, Kalrez;
All-PEEK tubing. Sample path is free of any metal parts
Aqueous channel wetted materials: PEEK, polycarbonate, Viton, stainless steel
Rated for aqueous solvents and alcohols
Wetted materials: PEEK, Kalrez
Metal-free channel wetted materials: PEEK, polycarbonate, Viton
Rated for organic solvents (e.g., THF, toluene, etc.)
Wetted materials: PEEK, stainless steel, Kalrez;
All-stainless steel tubing
Additional vapor sensor with interlock output
Organic channel: Upper channel block made from PEEK; Kalrez O-ring
Integrated in Agilent ChemStation, Thermo (Dionex) Chromeleon, and Shimadzu LCsolution
VOYAGER CDS™ stand-alone chromatography data system controls Eclipse, THERMOS[PRO] channel temperature regulator, Agilent HPLC modules and ASTRA software to initiate detector data acquisition.SCOUT DPS™ for virtual method development, method optimization and particle size analysis
|Display||Front panel LCD|
|Inputs||Autoinject In, Analog In|
|Outputs||Autoinject Out, Analog Out, Zero Detector, Pump Start, Agilent pump interlock (organic version only)|
|Dimensions||64 cm (L) x 36 cm (W) x 25 cm (H)|
Host PC requirements may be found in Computer Requirements.
Specifications subject to change without notice.
Other FFF Instruments
Eclipse™ AF4™ - Center-downstream injection FFF fluidics control system, optimal when use with HF5 or SEC is not important. The AF4 is required for the fritless channel and semi-preparative FFF.