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1 Designed for leading researchers working at the limits of conventional mass spectrometry capabilities who need to further characterize and define their samples the Waters SYNAPT High Definition MS (HDMS ) System offers unique, enabling functionality. Utilizing Waters patented Triwave technology, the SYNAPT HDMS System combines high-efficiency, ion-mobility based measurements and separations with high-performance quadrupole, time-of-flight (TOF) mass spectrometry. The SYNAPT HDMS System enables researchers to analyze samples that are differentiated by size, shape and charge as well as mass to deliver increased specificity and sample definition beyond that achievable by conventional mass spectrometers. PERFORMANCE SPECIFICATIONS LOCKMASS REFERENCE SPRAY T-WAVE ION GUIDE DRE LENS PUSHER DETECTOR QUADRUPOLE The SYNAPT HDMS System can operate in two modes: ANALYTE SPRAY TRAP ION MOBILITY SEPARATION TRANSFER (a) TOF mode (b) Mobility-TOF mode TOF Mass Resolution in Positive Ion 1 2 OIL-FREE AIR-COOLED TURBOMOLECULAR PUMPS SCROLL PUMP 5 10,000 measured on the (M + 6H) 6+ isotope cluster from bovine insulin (m/z 956) (b) W Mode 17,500 measured on the (M + 6H) 6+ isotope cluster from bovine insulin (m/z 956) TOF Mass Resolution in Negative Ion 10,000 measured on the (M - 4H) 4- isotope cluster from bovine insulin (m/z 1431) (b) W Mode 17,500 measured on the (M - 4H) 4- isotope cluster from bovine insulin (m/z 1431) Positive Ion MS Sensitivity The peak at m/z 556 from a solution of 50 pg/μl leucine enkephalin in 50/50 acetonitrile/water + 0.1% formic acid, infused at a flow rate of 5 μl/min, will have an intensity of greater than 1,700 counts per second. The instrument will be tuned to 10,000 resolution (as demonstrated on bovine insulin) and the mass range will be set to 1,000 Da. (b) EDC The peak at m/z 556 from a solution of 10 pg/μl leucine enkephalin in 50/50 acetonitrile/water + 0.1% formic acid, infused at a flow rate of 5 μl/min, will have an intensity of greater than 2,000 counts per second. The instrument will be tuned to 10,000 resolution (as demonstrated on bovine insulin), with sensitivity set to a maximum at 556 Da. Negative Ion MS Sensitivity The peak at m/z 503 from a solution of 500 pg/μl raffinose in 70/30 acetonitrile/water (no additives), infused at a flow rate of 5 μl/min, will have an intensity of greater than 1,800 counts per second. The instrument will be tuned to 10,000 resolution (as demonstrated on bovine insulin), and the mass range will be set to 1,000 Da.
2 Positive Ion MS/MS Sensitivity Using a [Glu] -Fibrinopeptide B solution of 100 fmol/μl, at a flow rate of 5μL/min and with the instrument tuned for 10,000 resolution (as demonstrated on bovine insulin), the intensity of the most intense y sequence ion from the MS/MS spectrum of the doubly charged precursor ion (785.8 Da) will be greater than 130 counts per second. This will correspond to a signal to noise ratio of greater than 50:1 (after a 3x9 smooth) on the most intense y sequence ion, for a 1 second scan. The instrument mass range will be set to 2,000 Da. Negative Ion MS/MS Sensitivity Using a solution of 500 pg/μl raffinose in 70/30 acetonitrile/water, at a flow rate of 5 μl/min and with the instrument tuned for 10,000 resolution (as demonstrated on bovine insulin), the intensity of the fragment ion at Da in the MS/MS spectrum of the precursor ion at Da will be greater than 130 counts per second. The instrument mass range will be set to 1,000 Da. Mass Scale Calibration Accuracy The mass measurement accuracy of the instrument in V-mode, using an internal lockmass, is such that the RMS error between the measured and the accepted masses of peaks which have sufficient intensity, and are free from interference from other masses, will be less than 2 ppm over the range Da. Mass Measurement Accuracy The mass measurement accuracy of the instrument, in W-mode, will be better than 2 ppm RMS, based on 10 consecutive repeat measurements of the [M + Na] + ion of raffinose (m/z ), using the [M + H] + ion of leucine enkephalin ( ) as the LockSpray lockmass. Analyte and lockmass peaks must have sufficient intensity and be free of interference from other masses. Mass Range The TOF mass range is ,000 Da in V-mode, and 20 26,500 Da in W-mode. The quadrupole mass range in non-resolving mode is 20 16,000 Da for a 4,000 Da quadrupole, and 20 32,000 Da for an 8,000 Da quadrupole.
3 Acquisition Rate Spectra can be acquired at a rate of 10 scans per second, except in pdre mode, when the maximum rate is 5 scans per second. Conditions: inter-scan time 20 ms, centroid or continuum, mass range 100 1,000 Da, single spray source. The sensitivity at 10 scans per second should be greater than 80% of the sensitivity at 1 scan per second. Dynamic Range The dynamic range, defined as the range of peak intensities that will give better than 3 ppm accurate mass (95% confidence) for 10 seconds of data, is at least 4 orders of magnitude, when measured on the m/z peak from leucine enkephalin. Isotope Ratios In the molecular ion isotope cluster of leucine enkephalin, the [M+H+1] + isotope will have an intensity of 33.8 (±1.0)%, the [M+H+2] + isotope will have an intensity of 6.9 (±1.0) %, and the [M+H+3] + isotope will have an intensity of 1.1 (±1.0) %, all measured with respect to the intensity of the [M+H] + isotope at Da. The peaks must be free of interference from other masses. Peak intensities must be below the DDTC (Digital Dead Time Correction) limit. High Mass Precursor Selection Applicable to instruments with 8,000 Da quadrupole only. The low energy MS/MS spectrum of m/z from a solution of 2 μg/μl sodium iodide in 50/50 isopropanol/water will contain only m/z and its fragments. The intensity of the largest fragment ion will be less than 5% of the intensity of the precursor ion. MS/MS data will be acquired over the mass range 100 8,000 Da, with collision energy of 10 ev. MALDI-TOF MODE PERFORMANCE SPECIFICATIONS Mass Resolution in Positive Ion (a) V mode 8,000 measured on the (m + H) + isotope cluster from insulin B chain (m/z ). (b) W mode 13,000 measured on the (m + H) + isotope cluster from insulin B chain (m/z ).
4 Mass Resolution in Negative Ion LOCKMASS REFERENCE SPRAY T-WAV E ION GUIDE DRE LENS PUSHER DETECTOR (a) V mode ANALYTE SPRAY QUADRUPOLE TRAP ION MOBILITY SEPARATION TRANSFER 8,000 measured on the (m - H) - isotope cluster from insulin B chain (m/z ). (b) W mode 1 2 OIL-FREE AIR-COOLED TURBOMOLECULAR PUMPS SCROLL PUMP 5 13,000 measured on the (m - H) - isotope cluster from insulin B chain (m/z ). Positive Ion MS Sensitivity Measured in The peak at m/z from 10 fmol of [Glu] -Fibrinopeptide B will have an intensity of greater than 30,000 counts acquired from an entire line through a sample well. The instrument mass range will be set to 2,000 Da. When this combined acquisition is smoothed (5 window, 1 number Savitzky Golay) and background subtracted the signal-tonoise ratio will be greater than 90:1 when compared to a region of the mass scale between 1,768 and 1,818 Da. Negative Ion MS Sensitivity Measured in The peak at m/z from 100 fmol of [Glu] -Fibrinopeptide B will have an intensity of greater than 30,000 counts acquired from an entire line through a sample well. The instrument mass range will be set to 2,000 Da. When this combined acquisition is smoothed (5 window, 1 number savitzy Golay) and background subtracted the signal-tonoise ratio should be greater than 90:1 when compared to a region of the mass scale between 1,768 and 1,818 Da. Positive Ion MS/MS Sensitivity Measured in V mode, Precursor Mass of The peak at m/z (y 9 ) from 10 fmol of [Glu] -Fibrinopeptide B will have an intensity of greater than 1,000 counts for an entire line through a sample well. The instrument mass range will be set to 1,620 (Precursor+50) and the collision energy and instrument conditions set such that the intensity of this peak will be greater than 40% of the intensity of the precursor peak. When this combined acquisition is smoothed (5 window, 1 number savitzy Golay) and background subtracted the signalto-noise ratio should be greater than 60:1 for the peak when compared to a region of the mass scale between 1,450 and 1,490 Da. Mass Measurement Measured in V mode, Positive Ion The mass measurement accuracy will be better than 2 ppm rms, measured from a mixture of PEG oligomers between 700-2,500 Da; using an internal reference peak and an instrument calibration that covers the same mass range.
5 MOBILIT Y-TOF MODE PERFORMANCE SPECIFICATIONS The Triwave cell is the enabling technology of the SYNAPT HDMS System. It is comprised of three main components: (a) Trap T-Wave Ensures high-efficiency by trapping ions prior to ion mobility separation; the trap can also operate as a collision cell. (b) IMS T-Wave Provides reproducible separation of ions based on their ion mobility. (c) Transfer T-Wave Transfers separated ions to the oa-tof for mass analysis; the transfer T-Wave can also operate as a collision cell. Mobility Functionality Separation of ions based on their mobility (size, shape and charge) Time Aligned Parallel (TAP) fragmentation High duty cycle (HDC) for extended detection limits Drift Time measurements: Record drift times between 33µs and 90 ms Control of Mobility Separation Control of Triwave height and velocity over linear ramps and user defined programs Mobility gas pressure control from ~10-4 mbar to 1 mbar (nitrogen) Provision for multiple mobility separation gas types Automated control of mobility conditions for routine operation Comprehensive manual control of mobility and Triwave operation for research use Waters is a registered trademark of Waters Corporation. HDMS, High Definition MS, Synapt, Triwave, and The Science of What s Possible are trademarks of Waters Corporation. All other trademarks are the property of their respective owners Waters Corporation. Printed in the U.S.A. January EN JH-PDF Waters Corporation 34 Maple Street Milford, MA U.S.A T: F:
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