Introducing the DSQII

Terry Jeffers

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AgendaUpdate on Thermos Environmental Initiatives

Discussion on new approaches to the challenges of 8270

Introducing Thermo Electrons Productivity Kits

EnviroLab Forms 2.0 High Throughput Quatification

New application note: Rapid Analysis of 8270

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Thermo Electron: Environmental InitiativeFocus on Environmental Laboratory needs:Requirement for Productivity

Need for good work flow in Generating, Reviewing and Reporting data

Benefit from guides for operation, maintenance and troubleshooting specific Methodologies

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Thermo Electron: Environmental Initiative Product Sales Specialist Environmental Demo Chemist Environmental Seed Chemist*New hardware: DSQ IIProductivity Kits:8270 and 5258260 and 524

Pitt Falls for Method 8270Those little things that stop everything

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Activity in the GC/MS systemActivityThese sites can be everywhereInjector, column, sourceSPCC compounds are used to show good systemNitrophenols are some of the best compounds to find active sitesMinimizing Effects of ActivityDeactivated linersGood column that shows no active sitesHigh flow rates to reduce the residence time of any active compound.Balance temperature zones to reduce activity and thermal break down.

ChromatographyHow important is it really?

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GC and AutosamplerSeparations accomplished with slow chromatographyCommon GC methods for 8270Fast injection250 - 300 0 C injector temperature1 mL/min flow rate30 m x 0.25 mm ID columnOven ramps between 5 0 and 10 0 C/min from 35 0 to 300 0 CSeparations accomplished with fast chromatographyAlternate GC method for 8270Hot needle injection250 0 C injector temperature3 mL/min flow rate30 m x 0.25 mm ID x 0.5 m columnOven stepped ramps to provide the critical pair separations.

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Hot Needle Injection: Run at Lower Inlet Temperature: Fewer Replacements of Liner

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Increase the Mobile Phase: 3 mL/min vs 1mL/minIncrease capacity of stationaryNo fronting of peak shape for 160 ng/uL PNA Decrease the irreversible absorption of active speciesPentachlorophenol < 10%RSD(5-160 ng/uL)0.1 ng/uL MDL replicates at 1 ng/uL 4.7%RSDMinimal breakdown for endrin or 4,4-DDTProvide Gaussian shape peaks for the most volatileNo solvent skewing of NDMA and pyridineShorter run times: 17 minutes

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Effects of increased flow ratesWhy increase the flow rate to 3.0 mL/minLearned from applications developed for the analysis of explosives.Decrease the residence time of the compounds in potential active areas.Decreases the run time without higher temperatures that could potentially damage thermally labile compounds.Do not need to expose the column to higher temperatures that will damage it.Provides better separations and narrower peak widths.This is balanced with a 20:1 split flow ratio.

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Injection: Choice of Liner

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PNAs ( 5 ng and 160 ng)

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NDMA and PyridineSplitSplitless

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Two Methods: Splitless and Split

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TRACE Ultra DSQ II GC/MS

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Source materials and geometryNew DuraBriteTM Source Open GeometryPassive CollectorExtends Dynamic RangeImproves RobustnessOptional Closed EI Ion VolumeConstructed of new proprietary material for enhanced inertnessProtects Lens 1 from ion burn

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New DynaMax XR Ion Detection System New Linear/Log Hybrid Electrometer

Extends output current to 70A Provides dynamic range to >106 No loss of response or precision at the lower sample/analyte levels

SGE ETP AF635 Electron Multiplier

Detector linear range output current >30A Simple snap out replacement of multiplier assembly

Expected ResultsResults I can get, you can get them too.

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Calibration CurveFirst page of the calibration curve reportAll SPCC and CCC requirements metAverage of all RSD calibration curves of 7.46%

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SPCCs (5ng to 160 ng min >0.05)

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CCCs < 30% RSD for Initial Calibration Curve

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Quantitated Data FileFirst page of the Quantitation ReportThis shows the quantitation of the 20 ng/uL sample for the calibration curveMeets the requirements of the internal standards and surrogatesMeets all ion ratio requirementsNo manual integrations were required

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Robustness: Split (2gr sediment/2mL methylene chloride + 25 uL diesel fuel)

How can we prove itProductivity Solutions

Productivity SolutionsIntegrating the DSQ II with Software, Methods, and Consumables to Get Productive, Quickly

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Productivity SolutionsProvide all of the necessary material to run the methodProven Thermo ColumnDeactivated LinersStandards Snap and shoot prediluted for entire method validationAll other consumables for about 2 weeks Provide all of the softwareEnviroLab Forms 2.0Interactive Reference CDAll of the methods, sequences, Validation data used to generate all of what you see here and in the manualsTwo Boxes full of materials, manuals, and CDs.

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Productivity Kit SOP manual for methodHow to GuideQuick Start GuideProvide consumables for running methodStandards, liners, septa, syringes, column,Instrument method, processing method, reporting packageCD deliverableSOPData from method validation Instrument methodProcessing methodReporting software with templates

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Pieces of the Productivity Kit

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Quick Start Guide

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8270C/D Productivity SolutionCustomers system can be taken from installation sign off to method IDLs in as little as 2 days!8270 is a value added product which will sell for ~ $3000 US Add on service provided by Thermo FSEsUS Field Service Engineers are being trained on the 8270 PSKits serve as a great tool for training aides for the lab

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Productivity Solutions are a Win-WinGood for customers Improves overall experienceIncreases satisfactionEases transition to Thermo GC/MSGives optimal performance for the DSQ IIGood for ThermoEasy to support these solutions

Helps the customer and support by establishing a known set of criteria to achieve success.Address the new lab environment less highly skilled users with higher throughput requirements

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Redefining the Way We Look at GC/MSProductivity Solutions target routine GC/MS applications to offer value to the customer

EnviroLab Forms v 2.0High Throughput Quantification

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EnviroLab Forms v 2.0An entirely new data processing and reporting system running as a layer over Xcalibur 1.4While targeted towards environmental laboratories and applications, it has been shown to be valuable for any laboratory performing high-throughput quantificationAdds a significant complement of capabilities that Xcalibur Ion ratio calculationsPeak integrationReportingIntegrated data review and reportingNo more batch reprocessing to re-generate reports of edited resultsFully automated while still allowing sophisticated manual review

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Data Review

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Report View

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EnviroLab Forms v 2.0Brand new workflow oriented user interfaceSubstantially improved ion ratio review and editing supportComprehensive set of textual and graphical reportsFully automated, fully manual or any combination in betweenCan be used as a high-throughput quantification engine for any applicationSimply turn off or dont use those options which arent relevant for a given user

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ConclusionThe DSQ II system provides updates to hardware and software that make it easy to useThe Productivity Solution for method 8270 provides a method that has been validated across multiple DSQ II systems.All of this allows a new operator to be productive immediatelyThe proven changes to the method allow an increased amount of productivity and throughput for the analyst and the laboratory.

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Questions?Thank You

Activity is always a problem. Activity seems to be everywhere and some compounds just seem to find those active sites. The 8270 method uses system performance check compounds (SPCC) to test for activity and to make sure the system will meet minimum performance criteria. Sources activity are the injector in general with the liner specifically, the column, and the MS source. The nitrophenols seem to be the best compounds to find active sites in a system.

The key to minimizing activity in the injector is a deactivated liner. The deactivation can be degraded by samples and high temperature. The column that is used should be able to separate different classes of compounds, acids bases and neutrals. This by itself is challenging but when you add matrix the problems increase dramatically. As a column ages not only will there be active sites that develop but the stationary phase will get damaged enough that the separation efficiency decreases to the point of being unusable. To minimize active site effects increased flow rates can be used. Higher flow rate are used to minimize the residence time, minimizing the time spent near active sites. This also has the effect of decreasing run time, providing narrower peaks, and better separations.

The injector also needs to be run at a temperature that balances the transfer efficiency of the PAH compounds with the potential of increased activity at the higher temperatures. These are two alternate GC methods that have been found to work with the DSQ II. The 300 C injector temperature should not be used with this method. It will cause too much degradation of the septum and provide no benefit for the analysis. Thermo GCs can volatilize the same material at a lower temperature setting because of the positioning of the heater, it provides a very even heating across the entire injector body. There are no relative hot or cold spots.

The hot needle injection was required to give the best results for all of the compounds in the list. It provides a consistent injection technique and does not give analytes a chance to remain condensed on a relatively cold needle.

The increased flow rate was seen to provide better results and will be demonstrated later. This shows the difference in some of the injection techniques that are available. The Hot needle injection allows you to set a lower injector temperature while decreasing descrimination for high boiling point compounds.Increasing the mobile phase provided superior results to the 1 mL/min flow rate. The capacity of the stationary phase increases and is able handle greater concentrations than it could otherwise. The effects of active sites that do remain in the system are greatly reduced. The most volatile compounds are separated from the solvent and baseline separated from each other. And lets not forget shorter runtimes. We have been developing applications for a long time for GC/MS. We got the opportunity to cross over some of that knowledge from one application area to a few others. We were able to take some of those techniques and apply them to other areas. There are many compounds in the 8270 method that have similar functional groups to explosives that are analyzed for in forensic applications. Increasing the flow rate higher than what is normally used in this method is one technique that proved very useful for method 8270.

There is no way to get rid of all potential active sites. Even if you did, after a few injections there could be enough damage to the system to cause active sites. The 3 ml/min flow rate pushes the compounds through the system fast enough to virtually eliminate any potential effects of active sites, even on a system that has been in use.The run time of the analysis is decreased without increasing the temperature ramp or the upper temperature of the GC oven. The compounds that decompose at high temperatures are able to be pushed out faster and are not subjected to even more heat.Since the column will not be exposed to higher temperatures and faster ramps the stationary phase will not be damaged. This increases the life of the column and the length of time between cleanings of the DSQ II.The peak shape becomes narrower, peaks become taller, and the software has an easier time finding and integrating peaks. This all adds up to increased throughput and productivity. It makes it easier on the analyst in time and effort for any one system. The 5 mm ID tapered liner was chosen for this analysis because it provides a large volume for the expansion of the sample once it is vaporized and provides the best protection for the metal parts of the injector. There is enough room at the bottom of the liner to allow for a split injection. In this case the split ratio used to validate this method was 20:1, this is actually a very high flow rate when it you remember that it is a 3 ml/min flow rate through the column that is being used. All of the other liners pictured were tried during the method development. None of them showed the best all around performance as the pointed out. Active sites were a major concern and were eliminated from the injector with this Thermo liner. New deactivation of Splitless 5 mm liner lower breakdown with wide bore liner than with 3 mm because the sample remains farther from the walls of the liner. Equal activity performance to press fit liner but easier to use and replace. When operated at 250C with hot needle injection, change once in 3 months of diesel injections. When operated at 275C, change 2-3 times a week. It was found that higher temperatures eventually destroyed the deactivation.

This slide shows that there is no difference in peak shape from the low point of the curve, 5ng, to the high point, 160 ng. There is a difference in retention time because of the increased amount of material that makes up the peak. The difference is 0.02 min or 1.2 seconds, a 0.11% difference.The circled peaks are NDMA and pyridine, respectively. The difference is the column and flow rates that are used. The top chromatogram is the 3 ml/min flow rate, that requires a split ratio of 20:1. These peaks are resolved from the solvent and baseline resolved from each other. This is the TR-8270 column 30m x 0.25mm ID x 0.5um film thickness, providing the best separations and fastest run times. The bottom chromatogram is the the TR-8270 column with 1.0um film thickness. This is a splitless method putting all of the sample and solvent onto the column, which requires the thicker film. It does not give as good of separations but may allow for lower detection limits, there is no splitting. If the thicker film is not used there is no way to separate the first 2 compounds from the solvent or from each other.These are the exploded views of the entire analysis with the different GC methods with the different columns. It is very important to match the column with the GC instrument method. If that is not done then there is the potential to overload the column, cause active sites, and not be able to meet the chromatographic QC of the method. The new detection system for the DSQ II maintains the ability to detect very small quantities while gives the ability to quantitate to very high concentrations. This can be seen in the calibrations for method 8270. It is possible to use either average response factor or a linear calibration curve for compounds even out to 160 ng/ul. This will save the operator time and effort by minimizing the number of manual integrations that may be required and it most certainly decrease the number of samples that need to be diluted so that concentrations are inside a calibration curve.SPCCs, used to check for activity in the system, easily exceed the requirements of the method. Even the compounds that cause the most problem, 2,4-dinitrophenol, more than double the method requirements. This means that more samples can be run before any maintenance needs to be done on the system. The CCC compounds easily meet the requirements for the initial calibration curve and the