Thermo Scientific TurboFlow® Technology:
An Alternative Separation Technique for Complex Matrices
- Allows direct injection of biological samples into an MS/MS system
- Simplifies sample preparation and increases sample throughput
- Reduces ion suppression versus protein precipitation (PPT) and solid phase extraction (SPE)
- Minimizes method development time by transferring methods to different sample matrices
Why TurboFlow Technology is Superior
TLX systems are optimized to use turbulent flow chromatography to separate analytes from biological fluids prior to MS/MS analysis. The Aria® software controls all of the system functions including the ability to inject, transfer, and elute analytes from the patented TurboFlow column in less than a minute.
The TLX system injects the sample directly onto a narrow diameter TurboFlow column that is packed with large particles. Using the principles of turbulence, diffusion, and chemistry, the small sample molecules are separated from the sample matrix in the TurboFlow column.
Turbulence Inside a TurboFlow Column
When the mobile phase flows through the TurboFlow column, high linear velocities are created, which are 100 times greater than what is typically seen in HPLC columns. The large interstitial spaces between the column particles and the high linear mobile phase velocity creates turbulence within the TurboFlow column.
Diffusion - Molecules Moving Through a TurboFlow Column
Since small molecular weight molecules diffuse faster than large molecular weight molecules, the small sample compounds diffuse into the particle pores.
The turbulent flow of the mobile phase quickly flushes the large sample compounds through the column to waste before they have an opportunity to diffuse into the particle pores.
Chemistry - TurboFlow Column Chemistry Binds Small Molecules and Large Molecules Flow to Waste
TurboFlow technology employs chemistry to further separate analytes from other sample molecules.
Of the sample molecules that enter the pores, those that have an affinity to the chemistry inside the pores, bind to the column particles’ internal surface. The small sample molecules that have a lower binding affinity quickly diffuse out of the pores and are flushed to waste. TurboFlow columns are available with a variety of column chemistries to accommodate different analyte types.
A mobile phase change then elutes the small molecules that were bound by the TurboFlow column to the mass spectrometer or to a second analytical column for further separation.
TLX Systems Simplify Sample Preparation by Dramatically Decreasing Steps
Assays that currently require complex sample preparation procedures can be simplified by using the TLX system. The dramatic time savings could make screening for compounds more practical.
Typically, the determination of CAH from bloodspots involves complex sample preparation. Using a TLX system, the sample preparation was greatly reduced, making CAH determination feasible as a screening test.
Run Multiple Matrices with One Method
A TurboFlow method for most analytes can be transferred to different matrices, eliminating time consuming method development and maximizing sample throughput.
TLX Systems Reduce Ion Suppression
When samples are processed on TLX systems, sample components that cause ion suppression are removed. In a study to examine ion suppression, no ion suppression was observed when injections of raw plasma were processed by the TLX system. When protein precipitated plasma or extracts of plasma from Solid Phase Extraction (SPE) were injected into the MS/MS system, dramatic decreases of the analyte SRM signal occurred as residual sample components eluted from the system.
TurboFlow Method Reduces Ion Suppression
Courtesy of Joseph Herman
When Protein Precipitation (PPT) is used to crash out proteins from plasma, there are still many components remaining in solution. This can cause ion suppression during analysis, resulting in a loss of signal. Some SPE methods can be efficient at removing matrix interferences but all too often, those methods vary from compound to compound and do not remove all of the interferences, also resulting in ion suppression. SPE often concentrates contaminates.
TurboFlow technology minimizes ion suppression for sample extracts, which is a significant advantage compared to SPE and PPT. As a result, it is the preferred method for cleaning biological fluids.