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Increase Mass Spectrometer Throughput up to 400%John FinkProduct Manager Aria&Transcend2Agenda Current trends of liquid chromatography-mass spectrometry(LC/MS)market Current solutions to solve todays most common problems Multiplexing technology and its benefits New LC/MS software feature to help make LC/MS even better3Todays LC/MS Market Mass spectrometer are a valuable piece of capital equipment Budgets for capital equipment are under strain Maximize productivity of mass spectrometer without increasing costs4Synopsis from Industry Research Report“After a period of expansive opportunities and extensive growth,the life sciences are facing an economic landscape that has fundamentally changed compared to the boom time of the late nineties and early years of this decade.”“Despite this grim reality,rays of hope emerge as many scientists turn to technology and innovation as a way to not only survive,but to also maintain their position or reputation in their chosen field during these tough economic times.”“Collectively,these organizations are alsoto various degreesexploring new policies and procedures that will not only help to reduce their costs,but also to enhance their capabilities.”“Of the scientists polled for this report,71%plan to adopt new technologies in 2008 with the remaining respondents citing budgetary concerns for their lack of innovation.Nevertheless,some have chosen to adopt new technologies in an effort to actually reduce costs in the lab”Report#09-001 March 2009,“Prospering in a Down Market:Strategies for Life Science Suppliers”BioInformatics,LLC5How to Maximize Productivity of LC/MS?Run more samples,streamline Get answers/data faster Reduce cost of capital equipment Reduce cost-per-sample Handle changing work demands Multiple method validations Multiple users per mass spectrometer Intuitive,intelligent softwareWithout sacrificing data quality and sensitivity6How to Maximize Productivity of LC/MS?Run more samples,streamline Get answers/data faster Reduce cost of capital equipment Reduce cost-per-sample Handle changing work demands Multiple method validations Multiple users per mass spectrometer Intuitive,intelligent softwareThroughputCostsFlexibilityEase of Use7Improvement Challenges Increase throughput Optimize method for speed,not quality Lower cost Purchase lower quality instruments Increase flexibility Increase complications and chance of errors Easier to use Become simple,less flexible8What is LC/MS?Liquid chromatography is a separation method based on the partitioning of sample compounds between a stationary phase and a liquid mobile phase.It is widely-used as a method to separate the analytes of interest not only from one another,but also from the rest of the sample components.The result is analytes are eluted into the detector separately allowing accurate data collection.Mass spectrometers are detectors with high sensitivity,accuracy and large dynamic ranges.9How can we increase the productivity of LC/MS?How can we increase the throughput of a mass spectrometer without sacrificing quality or incurring too much extra cost?Are there negative effects to increasing throughput?10Solution 1 Method Changes to Increase Throughput Situation Have an existing method that works,but want it to go faster Solution Optimize method for speed Example faster gradients,shorter equilibration or wash times Possible Challenges Retention time shifts Shortening of column life Other variables may cause data problems Example11Solution 1 Optimize by Shortening Method04 min.0 min2 min0 min2 min12Solution 1 Potential Optimization ProblemThe highest quality mass spectrometer data will come from having the highest quality LC method0 min2 minIncrease retention time-missed peak0 min2 minCarryover poor separation bad data13Solution 2 UHPLC to Increase Throughput Situation Have great HPLC method,but need higher throughput Solution Upgrade to higher pressure pumps and smaller column UHPLC What is UHPLC Pump technology able to support higher back pressures 600 bar or higher Smaller particle columns are more efficient,but increase back pressure Sub 2 micron particle sizeEnable separation in less timeHigher pressure pumps enable working with smaller particle columns14Solution 2 UHPLC Introduction Benefits of UHPLC Faster methods without sacrificing data quality Sharper peaks increase selectivity and peak resolution Reduced solvent usage Possible Challenges Capital investment of new pumps,injectors,values,tubing May have to replace entire system very expensive Selection of new column Method will have to be redeveloped,tested and re-validated15Solution 3 Multiplexing to Increase Throughput Situation Have reliable HPLC or UHPLC method(s),but need higher throughput Solution Upgrade to multiplexing16Solution 3 Multiplexing Introduction?What is multiplexing Connecting two or more LC systems to a single MS Stagger the LC methods Takes advantage of the idle time when the mass spectrometer is not collecting data During equilibration of one method,other method can elute to mass spectrometer17Solution 3 Multiplexing Introduction What is Multiplexing Connecting two or more LC systems to a single MS Stagger the LC methods Takes advantage of the idle time when the MS is not collecting data During equilibration of one method,other method can elute to MS Benefits of Multiplexing Increased throughput of samples more peaks,more data Increased utilization of existing mass spectrometer Existing HPLC or UHPLC methods can be upgraded to multiplex Lower cost than buying additional LC/MS system Just a new front end Take your existing best quality method and multiplex it18Solution 3 Multiplexing DiagramPumpColumnColumnColumnColumnPumpPumpPumpMass SpecAutosamplerSelector Valve18 279 3645cWasteBypassValve?How does multiplexing work?Diagram above is representative of a Thermo Scientific Transcend LX-4 system19Solution 3 Multiplexing Increases Throughput0 min4 min.System1MS Data20Solution 3-Multiplexing Increases Throughput0 min4 min.System1MS DataAcquisition WindowIdle TimeIdle Time21Solution 3-Multiplexing Increases ThroughputDiagram above is representative of a Transcend LX-4 system0 min4 min.System10 min4 min.0 min4 min.0 min4 min.System2System3System4The LC systems are staggered eluting stream is automatically introduced to mass spectrometer22Solution 3-Multiplexing Increases ThroughputDiagram above is representative of a Transcend LX-4 system0 min4 min.System10 min4 min.0 min4 min.0 min4 min.System2System3System40 minResultingDataThe LC systems are staggered eluting stream is automatically introduced to mass spectrometerIndividual data files23Solution 3-Multiplexing Increases Throughput Software automatically ensures mass spectrometer is dedicated to each individual elution stream Increases sample throughput and increases the utilization of the mass spectrometer without data or sensitivity lossDiagram above is representative of a Transcend LX-4 system0 min4 min.MS Data No MultiplexingMS Data MultiplexedAbove example increases mass spectrometer throughput by 400%0 min4 min.24Solution 3 Multiplexing Saves Money The mass spectrometer is a valuable instrument Want to maximize its utilization Buying an additional LC/MS systems to increase throughput costs more than a multiplexing LC system A multiplexing LC system saves up-front capital costs Also saves floor space and maintenance costs25Solution 3-Multiplexing Saves Money?Upfront capital costs and throughputCapital costs are estimates of typical MS and Multiplexing LC systems26Solution 3-Multiplexing Saves Money?Amortized sample costs27Solution 3-Multiplexing Saves Money28Solution 3-Multiplexing Saves Money600 samples/week1,200 samples/week2,400 samples/week29Solution 3-Multiplexing Saves Money30Solution 3-Multiplexing Saves MoneyThe additional cost of a multiplexing LC system is paid off in less than 3 months31Multiplexing Application Throughput EquipmentTranscend LX-4 System?4-Channel LCThermo Scientific Aria OS softwareThermo Scientific TSQ Vantage mass spectrometer ApplicationFour-channel LC separation for high-throughput,simultaneous analysis of a compound of interest for drug discovery Compound:1-minute method with a 15-second data collection windowSame method runs on separate channels,simultaneously Quadruple throughput32Multiplexing HTS,15s injections Throughput33Solution 4 Multiplexing Increases Flexibility Situation Need to run multiple HPLC or UHPLC methods,but dont have enough mass spectrometers.Or dont want to go through set-up procedures every time changing methods on a MS Solution Upgrade to multiplexing34Solution 4-Multiplexing Increases Flexibility0 min3 min.0 min3 min.System1Method 1Method 2Diagram above is representative of a Transcend LX-4 system35Solution 4-Multiplexing Increases Flexibility0 min3 min.0 min3 min.System1Method 1Method 20 minDiagram above is representative of a Transcend LX-4 systemResultingDataIndividual data files36Solution 4 Multiplexing Increases Flexibility Multiple channels enable flexibility in methods Each channel can be a completely different method Validate each method and run when needed Different methods can run simultaneously on different channels Can start at different times to increase throughput(walk up,load samples,run)Once set-up,no changes are needed-Just select method and run Saves considerable money and time-No system set-up or changes between methodsMultiple Methods on a Single MS=Most Flexibility37Multiplexing Application Multiple Methods EquipmentAria TLX-2 System?2-Channel LCAria OS softwareThermo Scientific TSQ Quantum Access MS ApplicationTwo-channel online extraction and LC separation for simultaneous analysis of opiates and benzodiazepines in forensic toxicology testing.Benzodiazepines:5.5-minute method with a 2.5-minute data collection window Opiates:7-minute method with a 3-minute data collection windowBoth panels run separate and simultaneously double throughput38Multiplexing Benzodiazepines Multiple Methods5.5 Minute MS Acquisition Window2.5 Minute MS Acquisition WindowSame Data,Same Quality39Multiplexing Opiates Multiple Methods7 Minute MS Acquisition Window3 Minute MS Acquisition WindowSame Data,Same Quality40Solution Review-Multiplexing Benefits Reviewed-throughput,cost,flexibility Possible Challenges Initial higher capital investments(Approx 15%-45%higher)Payback is less than three months in the example we covered41Multiplexing is Easy-to-Use Single software for all LC systems All channels,methods,samples and valves all controlled by a single software Intelligent recovery-If running same method on multiple channels and a channel fails,will move samples to operational channel Variables for method development I.E.Use a variable for a mobile phase concentration,run overnight,find best percentage Simplifies and quickens method development Same software runs 1-,2-or 4-channel multiplexing Directly interfaces with multiple mass spectrometer manufactures Do not have to write any custom softwareSoftware capabilities above are representative of Aria OS software42How to Maximize Productivity of LC/MS?Run more samples,streamline Get answers/data faster Reduce cost of capital equipment Reduce cost-per-sample Handle changing work demands Multiple method validations Multiple users per mass spectrometer Intuitive,intelligent softwareThroughputCostsFlexibilityEase of Use43Multiplexing Maximizes Productivity of LC/MS Run more samples,streamline Get answers/data faster Reduce cost of capital equipment Reduce cost-per-sample Handle changing work demands Multiple method validations Multiple users per mass spectrometer Intuitive,intelligent softwareThroughputCostsFlexibilityEase of UseWithout sacrificing data quality and sensitivity44New Aria OS software feature45Pressure Trace as a DiagnosticPicture with boundariesSet Upper Pressure LimitSet Lower Pressure LimitPressure traceScreenshot from Aria OS software46Pressure Trace as a DiagnosticPicture with boundariesCorrect Solvent BottlesSolvent Bottles ReversedScreenshot from Aria OS softwareResult software does not detect problem47Pressure Trace as a DiagnosticPicture with boundariesSet Upper Pressure LimitSet Lower Pressure LimitScreenshot from Aria OS softwarePressure trace48Pressure Trace as a DiagnosticPicture with boundariesCorrect Solvent BottlesSolvent Bottles ReversedScreenshot from Aria OS softwareResult software detects problem49Multiplexing as Part of Larger Workflow Automated mass spectrometer method development Parameters from data base Optimization/Acquisition LIMS in LIMS/data crunch out High throughput screening Open access50Questions.?51Contact Info John Fink 508-520-5500