MatrixEx, world most advanced matrix-elimination technology!
Every Chromatographer in Food Safety, Environmental, Water analysis, Pharmaceutical, Clinical, Toxicology, Sport Medicine, Forensics, Proteomics, Metabolomics, Trace analysis, Combinatorial and medicinal chemistry battling various degrees of matrix effect problems that lead to doubtful, non- reproducible results and bad reputation. MatrixEx is worlds first and only devise to remove matrix contamination, to renew and extent column life and to fully automate your chromatographic method so that you can concentrate your time on meaningful method development. MatrixEx is a ad-on device, ideal for enterprises that often have to change columns and to compete in the market. Productivity gains is the key to expand your wealth. MatrixEx is the ultimate productivity tool. It is the tool to compete effectively and efficiently with your procrastinating competitors using existing and new chromatography technology.
1. The Problem
Many samples comprise extracts of food, body liquids and crude from chemical synthesis. Even after careful pre-treatment the sample still consist of a small quantity of target molecules in a large quantity of matrix residue. (High ratio of matrix to target molecules).
Analytical chemists like to perform more analyses in a less time. They dont like to change columns / eluents often and like to employ robust, simple and cost efficient methods.
Most commercial laboratories work in a very competitive environment. To stay competitive most labs have to use modern equipment (e.g. LC-MS/MS) for 24 h day without much labour for three to five years so as to depreciate their acquisition before they are able to acquire the next generation of equipment.
The matrix effect is particularly strong in food safety analysis. Finding traces and metabolites from about 1000 different commercial pesticide on a large variety and quantity of foods is like finding needles in a hay stack. The samples are freeze dried, pulverized and extracted (liquid / liquid, SPE or QuETCHERS) with more or less labour intensive methods. The extract is injected in a LC - MS or LC - MS / MS to measure the masses of the pesticide residues.
With this strategy food matrix is carried into the chromatographic process. This leads to matrix elution and equipment contamination, to uncontrolled sample matrix co-elution, to non-predictable signal suppression, to erroneous results and to a bad professional image, possible claims and to pressure on job security.
Professional labs have to cope with a target range of approx. 300 to 400 pesticides that vary from very polar to very apolar. To cover the broad range of polarities requires a sample run over two or more different LC columns. This means high and growing labour cost, no process automation and sometimes wrong results. Food supplier may not agree with the data and to challenge the test results. This leads to stress, court cases and in a worst case scenario to loss of your business! Many undertake appropriate measures to reduce risk. Some acquiring acquire new, very advanced technologies and some smarten up their existing technologies. There are a number of choices to address this problem but MatrixEx is the most competitive and most advanced technology.
2. What tools are available already?
The matrix problem is large and has induced many research projects in all countries. Today we can choose from hundreds of publication and methods just to mention three alternatives
Problem solving strategy 1.
On-line sample preparation strategy: Through-put can be increased by merging sample preparation with chromatography. Unfortunately, this method is still a one-dimensional, sequential system!
Problem solving strategy 2.
Multidimensional Analysis strategy: e..g. Agilent Stream Select LC / MS solution. Multidimensional Analysis is known for many years and has reached its limits.
Problem solving strategy 3.
A few public research institutions have started to procure High Resolution MS and to dilute their samples so as to reduce signal suppression. They argue that this is the future strategy for heavy contaminated samples.
- With dilution one reduces ion-suppression only partially: In matrix loaden samples this will not solve the problem.
- This strategy will still accumulate (the more retentive) matrix on the column, which later will elute, contaminate the MS and interfere as usual.
- This strategy requires very sensitive MS instruments - such instruments are very expensive. Routine analytical labs have to compete commercially. They cant afford to procure the most sophisticated instruments for routine analysis. Very few organisation can afford up-market instruments.
3. Search for better technology
In a competitive environment what matters is productivity. Above mentioned strategies dont improve productivity. We have to address the sample matrix to pesticide residues ratio intelligently and we have to look forward in time.
In the global world we will have more competitors, a wider range of pesticides and more companies that participate in the market with faster or low cost services. Some new competitors will challenge the current players to grab some market share. Regulation authorities may demand more validated results In short the world will become more complex and analytical labs will be challenged to demonstrate their ability to cope with the growing complexities.
Complexities cant be addressed with highly tuned single purpose problem solving technologies e.g. to increase sensitivity of MS. The very best Mass Spectrometer can’t do a proper job if the upstream sample preparation and LC system is not working properly! MS separate ions, LC separate molecules according to their structures. Both are important to cope with growing complexities.
In hyphenated technologies we have three strategic choices
1. To increase efficiency of the HPLC - side
2. To increase efficiency of the MS- side or
3. To increase effectiveness and efficiency of both.
In the past few years tremendous progress has been made in advancing the MS side. Modern quadrupol instruments can detect several hundred MRM transitions / minute, which should allow the simultaneous detection of hundreds (or thousands) of compounds in one chromatographic run (without compromising sensitivity)! During the same time there were also considerable improvements achieved on the HPLC side. Unfortunately both improvements have not advanced together as a unit. Today system suppliers compete with other system suppliers and component suppliers. Unfortunately for the users they have started to hinder access to their system soft- and hardware. Concerned chromatographers have formed interest groups which demand “open access” from the suppliers.
In complex processes advancement is achieved by by advancing both sides through additional technologies. Kamasutra is about 400 year old book packed with wisdom about achieving a balanced, satisfied and sensuous life between couples or groups. and to do things with more passion. It teaches harmony between partners and to do things passionate. Kamasutra is a tremendous source of inspiration for advancing effectiveness and efficiency in hyphenated technologies. It inspires partners to experiment together to enhance life for both. This will lead to a more aware and fulfilled life. Kamasutras wisdom urges us to go beyond the obvious, to open-up our perspectives and to extend our skills in collaborating with other partners. Kamasutra is an inspiration for a paradigm shift in hyphenated technologies.
4. Birth of the ultimate productivity tool
Many new analytical strategies have been and will be created and evaluated including LC-MS, GC-MS, Immuno-Assay, use of biosensors, LOC, and some more will emerge. Ultimately, Pesticide Analysis is about guaranteeing food safety reliably and cost effectively. Governments initiate, support research and creation of new technologies. As time goes by the governmental support will be abandoned. Industry will takeover the sustainable new technologies to “industrialize” them. The rest will disappear.
Industrialization means that commercial enterprises take over the process. There will be companies that offer Pesticide Analysis as commercial service. Commercial organisation are in competition with each other. Those that are productive survive. They offer fast, precise, reproducible and a diverse range of relevant data for a very competitive price. The most productive enterprises are the winners in this race. Based on that background logic we decided to create intelligent tools for our clients so that they can provide the most productive analytical service to their clients.
2010 Dr. Manol Roussev and Mr. Jörg Plagge in the pesticide analysis labs of Wessling GmbH developed a technical concept to reduce sample matrix contamination together with column switching specialist MayLab Analytical Instruments GmbH. Based on the “Kamasutra Inspired Philosophy” (see KIT ) it was clear not to reduce but to improve process diversity and system productivity. Liquid chromatography is able to separate molecules according to shape, hydrophobic and hydrophilic interaction, ionic interaction and chiral interaction. MS can separate ions and measure masses.
What had to be done was to create a device in between existing LC and the MS that improves the productivity of both components and make it possible to extend process and detection parameters to create and measure additional dimensions. Maylab had great support from a number of MS manufacturers and friends from academia to get access to their MS software. A number of prototypes were developed and tested in Wessling laboratories. The team was extended with a third partner, LCC Engineering & Trading GmbH. LCC is responsible for global marketing, sale and physical distribution of MatrixEx.
As team we are adamant and passionate to create the most productive tool for eliminating matrix contamination in analytical processes. The key challenge was
- To eliminate matrix contamination visa traces of a wide range of contaminants or target molecules
- To be competitive.
5. Matrix Contamination is well researched!
Many Researcher investigated the composition and elution of the various matrices. Kittlaus showed that under appropriate chromatography conditions the majority of target molecules are leaving the column prior to the matrix molecules.
Flushing the column from matrix is lengthy and cumbersome.
Others started to experiment with various strategies to reduce or eliminate matrix contamination in the sample preparation step. Plagge/Roussev commenced testing of different sample matrices and could demonstrate with matrix effect profiles using QuECHERS that signal suppression has an impact of 80% and more . We could show that quantification of the measured concentrations due to signal suppression in the chromatographic separation can be compensated mathematically through the so-called post column infusion .
Based on the technique of post-column infusion we could measure experimentally, that by the end of the chromatographic separation sample matrix to remain on the HPLC column. Even with a significant prolongation of the final washing process using a pure organic eluent (e.g. 100% methanol) we discovered that even after 60 minutes of elution there was still remaining sample matrix on the chromatographic material (see Fig. below).
Above fig shows injection of QuECHERS extract of food products to contain heavy matrix. Washing the column is necessary. MS measuring time and washing time is lost time!
6. Evolution of MatrixEx the ultimate matrix elimination technology
Base on the observation we concluded the most successful strategy is to have an alternating chromatography process and column flushing capabilities as well as a virtually "unlimited" columns selection, combined with a column switching system that allows a free choice of columns and without restriction of direction of the eluent flow. If we run the column washing process in flow direction than the remaining sample matrix has to be flushed over the whole column. If we wash the column in reverse flow direction the remaining sample matrix is eluted from the column in a given time.
Using the MS software as component within MatrixEx enable us to detect the target molecules and to switch column and flow direction at the appropriate time.
The matrix elimination was further enhanced by controlling the chemical-engineering parameters through smart software such as eluent temperature with inbuilt column ovens, flow rate and operating pressure. In addition to that we can also utilize columns parameters and chemistry including eluent chemistry and composition in a very controlled manner. For routine operation we can automate the whole LC-MS process so that the machine will run over night or weekend.
The Process and its component were then submitted for patenting.
MatrixEx is the Swiss Knife in matrix elimination technology. MatrixEx that can be tuned to very many applications and to your particular environment.
7. Performance of MatrixEx
MatrixEx comprises of:
What you can do
- Conecting multiple columns to one MS (you increase MS productivity up to 40% and save lots of money!)
- Running two columns simultaneously
- Changing flow direction
- Robust high performance VICI switching or/and selection valves
- Standard 1/16 or micro bore (1/32) connector option
- Fast switching cycles and time(<100msec)
- Full bi-directional digital valve control
- UHPLC possibilities(low extra-column dead volume!)
- 5000 or 15000 psi pressure version
- Proprietary low volume multi-connector distributor
- High speed,high resolution stepper motor direct-drives
- Electronic calibration / re-calibration of valve positions
- You have full control over valves and oven
- Valve/oven controls are integrated into MS software platform
- Graphic overview for visualisation of valve /oven control
Column Switching & Thermal unit
What you get
Column switching unit
- After every process you start with new and clean columns (no Matrix contamination, longer column life)
- You are in control over configurable parameters
- Intelligent and integrated software
- Modern and functional design
- UHPLC compatible
- IPR protected
Thermal control unit
- Heating / Peltier cooling columns from 5°C to 75°C (Thermal focussing Chromatography)
- High temperature stability (0.1°C)
- High heating speed (10°C/Min) and cooling speed(3°C/Min)
- Wide oven space
- Full bi-directional digital oven control
- Graphical software control integrated into MS software
8. Interested in MatrixEx?
We are looking for
- Application clients with existing or new LC-MS facilities
- R & D Staff in Proteomics , Metabolomics, Natural products, Drug Discovery, Medicinal or Clinical chemistry with interest to accelerate and improve their process
Our “No Risk” offer
If you are seriously interested in MatrixEx
- Please contact us to discuss your current set up and goals.
- We then give you an estimate to availability and costs
- If our conditions are of interest we invite you to run your current method in our Application lab in Berlin
- The resultant data will enable you to estimate your productivity gain
- Based on the gathered data we will construct and sell MatrixEx to your specification
9. Process Description in Detail
To meet constantly increasing demands of today's consumer protection requirements, the technical performance of modern instrumental analysis must be continuously developed and improved. Complex coupling techniques such as GC-MS (/ MS) or LC-MS / MS are now established in every modern analytical laboratory. The requirement for higher sensitivity presupposes that the mass spectrometric detectors have a very high selectivity and specificity so that traces of analytes can be determined safely and reproducibly. The available instruments on the market from various manufacturers show excellent performance with respect to sensitivity and robustness. Particularly LC-MS have made remarkable technical progress in recent years.
With complex samples such as in pesticide residue analytics using LC-MS/MS we have often co-eluting sample matrix competing with the analytes. This leads to signal suppression or to greatly reduced resolution. We can’t be sure that after a chromatographic run and longer period of column purging no further matrix will elute to affect the quantitative analysis. The solution for achieving an efficient and robust trace analysis is to keep as much as possible from the sample matrix away from the MS. To eliminate th LC component is possible only in routine analysis of single compounds.
9.1. Productivity improvement Strategy 1
We can increase speed through using multiple columns in tandem mode combined with intelligent column switching!
9.2 Productivity improvement Strategy 2
Increase production speed through column switching and back flushing column. This will reduce the MS-idle time!
9.3 Productivity improvement Strategy 3
Using two columns in parallel (in alternative mode) with back flushing column
9.4 The System
The MatrixEx System is placed between your LC and your Detector system(MS UV etc.) to join both ends. The system comprises of the valve switching unit, the Thermostat (uniquely constructed Peltier oven) for up to 10 columns and a sophisticated software to program and run the desired methods automatically.
With the innovative valve switching system comprising of valve No 1 to valve No 6 we can chose any two columns independent of each other and whatever flow direction to be connected to the HPLC flow of the analytical pump or to the back flush flow of the wash pump.
You are in control of all variables in chromatography, flow rate, pressure, time, temperature, column chemistry and column physics.
9.5 Practical Performance Validation Data
System robustness and reliability are important validation criteria for all analytical systems. We use very high quality valves  driven by a motor that has faultless track record of more than 10 years to assure system reliability.
To achieve highest level of automation the system drivers are integrated into input software of the analytical system. This makes it possible to engage different measuring methods in relation to column phase and eluent composition and to run the system unsupervised e.g. overnight or over weekend. This results into a tremendous gain in equipment productivity.
Reproducibility with back flushed columns
Reproducibility without backflushed columns
The difference with regards to reproducibility of a hops sample extract is particularly good with regards to peak area of a deuterated internal standard used in routine analysis as seen below. The benefit become clear if we compare the percentages of standard deviation for each median (when using back flushing = 0.79% , in the absence of back flushing = 6.7%). It becomes obvious that uncontrolled sample matrix remaining on the column will produce significant variations due to the resultant signal suppression. Are both extremes compared the result is a difference of nearly 40% !. The advantages of the back-flushing with respect to "uncontrolled" signal suppression are clearly verifiable. The percentage deviation of the extreme is with backwashing less than 5%.
9.6 System Robustness
Robustness of the analytical system after 10, 3000 and 8000 injections.
Reproducibility after 8500 Injections
Continuous sample matrix removal will increase column life time substantially (up to 5 x)
MatrixEx software becomes an integrated part of the MS Software e.g. Sciex Analyst Software
Screen shot of Software
10. Summary of benefits using MatrixEx
We offer a novel column switching system that eliminates negative influences caused by sample matrix contamination. Installing MatrixEx between your LC and detector system will significantly increase quality of your analytical data and the efficiency of LC-MS / MS system. Aspects of innovation are summarized as follows:
- The specially developed column switching technology assures nearly complete removal of the matrix from the stationary phase after each chromatographic run including a regeneration of the HPLC column after the chromatographic separation.
- Chromatography and cleaning take place alternately without loss of time (equilibration occurs in parallel) with the use of identical stationary phases.
- The MatrixEx system achieves a significant increase in efficiency by automatically switching between different mobile and stationary phases in free selectable bi-directional flow (full automation). T
- The free choice of the flow direction prevents limitations in using "not flushable" HPLC columns.
- Compared to employing a traditional HPLC column thermostats with MatrixEx there is no manual column and eluent change necessary when different measurement methods are employed. Column and eluents switching can be automated.
- Automating the system combined with column pre-equilibration for various HPLC analysis methods the dead time between two methods is eliminated
- Integration of the equipment control will ensures total compatibility with LC-MS / MS systems including user friend handling
- Universality of the column switching technology allows analysis of complex sample matrices in the areas of food and environmental analysis, as well as toxicological and pharmaceutical studies.
- The professional final version of the switching technology is in routinely in use since 2012.
Please contact us to discuss your situation and product supply click here!
 H. Stahnke, T. Reemtsma, L. Alder. Compensation of Matrix Effects by Post Column Infusion of a Monitor Substance in Multiresidue Analysis with LC-MS / MS. Anal. Chem. 2009, 81, 2185-2192
 S.Kittlaus, J. Schimanke, G. Kempe, K. Speer. Assessment of sample cleanup and matrix effects in the pesticide residue analysis of foods using post-column infusion in liquid chromatography-tandem mass spectrometry. J.Chromatogr. A, 2011, 1218, 8399-8410
 M. Roussev, J. Plagge. From the injection to the test result - new aspects, 5th Berlin LC-MS / MS Symposium, 2013,