SOW PegasusHRT 4D.pdf

SOW and Specs

The Food Science and Market Quality & Handling Research Unit in Raleigh, NC requires a comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry system to replace the aging system that will be obsolete in the coming year (end of 2021). This instrument is necessary to ensure continuity of research as the analytical platform for metabolite profiling studies in pickled vegetables, peanut, and sweetpotato, and is required to complete the mission critical studies planned for two NP306 project plans. High mass accuracy and software to enable the identification of unknown compounds is a critical element in the new system, since ~80% of the compounds detected with our existing instrument have not been previously identified. This additional feature is required to advance our current capabilities to identify the full complement of small molecules that impact food quality and health-promoting potential.

The required capabilities of the system are listed below:

Overall System Requirements

• The proposed system must be fully integrated to allow for the control and operation of Gas Chromatograph (GC), comprehensive two-dimensional chromatography (GCxGC), Mass Spectrometer (MS), and data processing system from a single software platform.

• Must be fully compatible with an autosampler capable of liquid handling and injection, static headspace, and solid phase microextraction (SPME) obtained through a simultaneous separate purchase.

GC Specifications

• The GC column oven should have an operating temperature range from 5 °C above ambient to 450 °C.

• The GC column oven should provide a temperature set point resolution of at least 1 °C.

• The GC column oven should provide a temperature ramp rate of up to 120 °C /minute.

• The GC column oven should allow at least 20 temperature programming ramps in a single GC method.

• The split/splitless capillary injector should be equipped with electronic pressure and flow control.

• The split/splitless capillary injector should provide real time ambient temperature and pressure compensation.

• The split/splitless capillary injector should provide an automatic shut-down for hydrogen carrier gas when a pressure or flow measurement indicates the possibility of a leak.

• The split/splitless capillary injector should have a maximum split ratio of 7500:1.

• The split/splitless capillary injector should have a pressure range from 0 to 150 psi.

• The split/splitless capillary injector should have a pressure set point resolution of 0.01 psi.

• The split/splitless capillary injector should have a total flow setting range of 0 to 1250 mL/minute.

• The split/splitless capillary injector should have a maximum operating temperature of 400 °C.

• The split/splitless capillary injector should accommodate GC columns from 0.05 mm to 0.530 mm ID.

• The GC should allow communication through an RS-232 connection and provide at least two analog output channels.

• The GC should be equipped to communicate through a LAN interface.

• The GC should provide built-in diagnostics and comprehensive self-test.

• The GC should provide power-fail memory protection.

• The GC should be able to be operated in an environmental range from 20 °C to 35 °C, and 5 to 95% relative humidity.

GCxGC Specifications

• The GCxGC system must be capable of performing comprehensive GCxGC on the entire sample.

• The GCxGC system must be fully integrated, providing control of the GC, thermal modulator, secondary column oven, detectors, GCxGC data processing software, liquid autosampler, static headspace autosampler, and SPME autosampler from a single software platform.

• The GCxGC modulation system must be a thermally-based, quad-jet, dual-stage design, with two separate cold jets and two separate hot jets.

• The GCxGC thermal modulator cold jets should be liquid nitrogen cooled.

• The GCxGC system must be capable of detecting 1 pg of octafluoronaphthalene (OFN) injected and analyzed in GCxGC mode.

• The GCxGC system should have a peak tailing factor of less than 1.5 for undecane methyl ester.

• The GCxGC system must have a second dimension retention time reproducibility of no greater than ±10 milliseconds.

• The GCxGC system must be capable of measuring a chromatographic peak width of 60 milliseconds.

• The GCxGC system should be capable of modulation periods (second dimension retention time) from 1 to 65 seconds.

• The GCxGC system must be compatible with and include an automated, software-controlled liquid nitrogen leveler to facilitate unattended overnight operation.

• The GCxGC system must be compatible with both electron ionization and chemical ionization sources.

• The GCxGC thermal modulator must be capable of effectively (and quantitatively) modulating analytes within the range of C4 butane and C40 n-alkane.

• Alignment of columns through the GCxGC thermal modulator should be achieved by simple column guide orientation. The column set should not be required to be mounted or attached to a support board or similar structure.

• The GCxGC system must have an isolated secondary oven that can be controlled and programmed directly through the main data system. The secondary oven must be independently programmable from the main GC oven to enable adjustment of second dimension retention times to reduce/eliminate coelutions and wraparound.

• The GCxGC system must be capable of controlling the modulation period, hot pulse time, and cold pulse time in variable steps throughout the run (variable modulation).

• The GCxGC system must be capable of calculating and providing a corrected constant flow mode based on the column set and temperature zones used for the GC method.

GCxGC Data System

• The GCxGC data system must include a complete data processing, acquisition and reporting software package.

• The GCxGC data system must provide automated peak finding (deconvolution) of GCxGC data.

• The GCxGC software must provide an automated mechanism for determining analytes that have been modulated into more than one second dimension chromatogram and combining these peaks into a single analyte entry in the peak table.

• The GCxGC data system must provide automated peak area calculations based on automated combination of modulated peak slices.

• The GCxGC data system must provide automated analyte quantification of GCxGC data.

• The GCxGC data system must provide analyte calibration software that can generate single and multi-order calibration curves.

• The GCxGC data system must provide the ability to construct reference tables of known sample types.

• The GCxGC data system must provide the ability to automatically compare samples to the reference tables to quickly produce filtered lists of differences between the samples as well as similarities.

• The GCxGC data system must provide automated data export in a variety of electronic formats including ANDI, mzmL, and CSV formats.

• The GCxGC data system must provide fully integrated GCxGC hardware support for both liquid nitrogen or cryogen free thermal modulation.

• The GCxGC data system must provide 3D GCxGC views (contour, wireframe, and surface).

• The GCxGC data system must provide a GCxGC Column Calculator which determines the length of the secondary column based on first dimension column and flow characteristics.

• The GCxGC data system must classify regions of 3D GCxGC views for subsequent data processing.

• The GCxGC data system must provide user scaled mass defect plot of deconvoluted GCxGC spectra.

• The GCxGC data system must provide user scaled mass defect plot of raw (caliper) GCxGC spectra.

• The GCxGC data system must provide an automatic control switch to collect GC/MS data while setup for GCxGC (turn off modulator).

• The GCxGC data system must provide view of the different sections of the GCxGC method (columns, inlet, temperature zones, GCxGC parameters) within the software (tabbed view).

• The GCxGC data system must provide view of application windows on two computer monitors for easier data review.

• The GCxGC data system should provide graphical highlighting when the contour plot is zoomed in to look at a particular region of the chromatogram, the GCxGC system software should provide an optional full contour chromatogram overview insert that highlights the zoomed region for easy operator orientation in the data.

MS Specifications

• The mass analyzer must be capable of routine and sustained acquisition with a mass accuracy of 1 ppm or better. This will be in full mass range mode and be independent of acquisition speed or mass range.

The mass accuracy should typically apply to any m/z, from a S/N of greater than 100 to a point below saturation with standard data processing methods.

• The stability (RMS) of mass accuracy should be <1 ppm for at least 30 minutes at 10 - 200 spectra per second.

• To accommodate the capability for performing GCxGC and ultra-fast GC, the system must be capable of acquisition at not less than 200 full mass range, written-to-disk spectra per second. The full range of acquisition speeds must be 1 to 200 spectra per second.

• The mass analyzer must be capable of acquisition with a resolving power of 25,000 (fwhm) or greater at m/z = 219 (218.985) while acquiring a full mass range spectrum across the range of 10 to 1500 m/z.

The resolving power of ≥25,000 must be available and consistent independent of acquisition speeds and mass ranges.

• The mass analyzer must be capable of acquisition with a resolving power of 50,000 (fwhm) or greater at m/z = 219 (218.985)) while acquiring a spectrum of the range of 200 to 600 m/z. The resolving power of ≥50,000 must be available and consistent, independent of acquisition speeds and mass ranges.

• The mass spectrometer must be capable of ionization using electron ionization to generate traditional library searchable spectra.

• The EI source must have a dual filament design with automatic software selection of the other filament if one should fail during analysis.

• The mass spectrometer should be capable of analyte ionization using chemical ionization to generate pseudo-molecular ions.

• The system must have a mass range of 10 to 1500 m/z. Acquisition across this full range must not compromise system performance with respect to spectral resolution and mass accuracy.

• The system must be capable of detecting 100 fg or less of octafluoronapthalene (OFN) injected on column. This will be achieved in a full mass range mode in both GC and GCxGC acquisition modes.

• The mass spectrometer must be compatible with helium and hydrogen (optional) as carrier gases.

• The EI source should be designed to handle GC carrier gas flows ranging from 0.2 to 2 mL/minute.

• The system must have the capability for helium consumption control through GC and/or MS functionalities (GAs-saver mode).

• The system should have greater than or equal to 4 linear orders of dynamic range for quantitative analysis in GC mode.

• The system should have greater than or equal to 3 linear orders of dynamic range for quantitative analysis in GCxGC mode.

• The system must have a spectral dynamic range of at least 105 (100,000) to allow for analysis and detection of isotopes and low level analytes in the acquired spectra, including trace analytes in the presence of high level analytes.

• The MS should consist of electron ionization (EI) source, focusing optics, Time-of-Flight (TOF) mass analyzer, microchannel plate detection system, and high-speed data collection system.

• The MS should be differentially pumped between the ionization source and mass analyzer with a vacuum system consisting of a turbomolecular pump mounted on the source region with a 360 L/second (He) or 350 L/second (H2) pumping speed, and a turbomolecular pump mounted on the mass analyzer with a 440 L/second (He) or 420 L/second (H2). The turbomolecular pumps are backed by a single roughing pump with a minimum of a 75 L/minute (4.5 m3/hr) pumping speed.

• The MS system should have 2 differing levels of MS calibrant which are controlled by the software.

• The MS must include electronics and software to automatically correct for ion arrival time deviations and preserve spectral integrity.

• The MS should be able to be operated in an environmental range from 20 °C to 26 °C, with 20 to 80% relative humidity.

• Raw (caliper) mass spectra must not be skewed by the MS.

GC-MS Data System Specifications

• The GC-MS data system should be compliant with all Windows® 10 based operating systems.

• The GC-MS data system should provide automated overall system optimization algorithms that maximize signal intensity, maximize mass resolution, and calibrate the mass axis.

• The GC-MS data system must allow for the addition of samples to an acquisition table and/or reordering of samples in the acquisition table without requiring the analyst to terminate the sequence.

• The GC-MS data system should provide integrated control of the GC.

• The GC-MS data system should provide integrated control of the autosampler.

• The GC-MS data system should provide the ability to prepare analyst-defined quality assurance protocols for system optimization, vacuum leak tests, and detector response correction.

• The GC-MS data system must provide the ability to start acquisition via contact closure.

• The GC-MS data system should allow the analyst to schedule the quality assurance tests on a time basis, a per-sample basis, or both.

• The GC-MS data system should use the NIST library search algorithm for comparing sample mass spectra to databases of mass spectra and must include the most recent version of the NIST mass spectral library.

• The GC-MS data system must allow the creation and use of analyst-defined accurate mass libraries of mass spectra to be used for library search.

• The GC-MS data system should allow multiple libraries to be automatically searched simultaneously.

• The GC-MS data system should allow for an analyst-defined number of library matches to be returned from the library search algorithm.

• The GC-MS data system must provide the ability to calculate retention indices for analytes found in a sample.

• The GC-MS data system must allow for fully automated qualitative analysis consisting of: location of all peaks that meet analyst-defined criteria; deconvolution of the mass spectra for all located peaks to remove background-, matrix-, or coeluting-analyte signal; and tentative analyte identification by library search.

• The GC-MS data system must provide a fully automated peak find algorithm capable of detecting coeluting analyte peaks, analyte peaks buried beneath the background of the total ion current chromatogram, or analyte peaks buried beneath significantly larger concentration analytes and/or matrix interferences.

• The GC-MS data system must provide a fully automated mass spectral deconvolution algorithm that removes background signal, removes interfering signal from matrix or other analytes, and accurately proportions shared signal found in both the analyte spectrum and the coeluting interference resulting in a clean, full-range mass spectrum for more accurate analyte identification.

• The GC-MS system must allow for the presentation of deconvoluted mass spectra in coelution situations where the ion of interest is shared. The deconvoluted peak shapes should represent the portion of shared signal contributed only by the specified analyte in the coelution.

• The GC-MS data system should allow for a reverse search of a peak table for a target spectrum to see if the analyte is present anywhere in the sample.

• The GC-MS data system should allow for the automated comparison of two samples to generate a list of analytes found only in sample A, a list of analytes found only in sample B, a list of analytes found in both samples A and B, and a list of analytes found in both samples A and B but outside an analyst-defined concentration range.

• The GC-MS data system must allow for fully automated quantitative analysis based on single or multi-order calibration curves.

• The GC-MS data system must allow for all analyte information, including mass spectral information, and all sample information to be exported as CSV file.

• The GC-MS data system must allow for ANDI, CSV, and mzml export of raw data.

• The GC-MS data system must allow users to choose the mass scale of mass defect plots on both raw and deconvoluted data.

• The GC-MS data system must allow searching ChemSpider from within the system to facilitate identification of unknowns.