The IMS/MS studies were executed with two different drift tube IMS (DTIMS)/MS platforms. The first was an in-house home-built DTIMS/MS instrument that coupled a 1 m IMS drift tube with an Agilent 6224 TOF MS upgraded to a 1.5 m flight tube (providing MS resolution of ~25 000),23 (link) and the second was the Agilent 6560 IM-QTOF MS platform (Agilent Technologies).24 (link)–26 (link) The Agilent 6560 was outfitted with the commercial gas kit (Alternate Gas Kit, Agilent) and a precision flow controller (640B, MKS Instruments) to allow for real-time pressure adjustment based on absolute readings of the drift tube pressure using a capacitance manometer (CDG 500, Agilent). For the DTIMS measurements on both instruments, ions were passed through an inlet capillary, focused by a high-pressure ion funnel, and accumulated in an ion funnel trap. Ions were then pulsed into the drift tubes filled with ~3.95 Torr of nitrogen gas, where they traveled under the influence of a weak electric field (10 to 20 V/cm). Ions exiting the drift tube were refocused by a rear ion funnel prior to TOF or QTOF MS detection, and their arrival times were recorded.
Cdg 500
Manufactured by Agilent Technologies
The CDG 500 is a compact, high-performance vacuum gauge controller designed for a wide range of vacuum applications. It provides accurate and reliable measurement of vacuum pressure across multiple pressure ranges. The CDG 500 is compatible with various types of vacuum sensors and can be easily integrated into various vacuum systems.
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2 protocols using cdg 500
1
Dual DTIMS-MS Platforms for Ion Analysis
2
Quantitative Biomolecular Interactions via DTIMS-Q-TOF
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The sample solutions were injected at a 10 µM strand concentration on an Agilent 6560 DTIMS-Q-TOF instrument (Agilent Technologies, Santa Clara, CA, USA), equipped with a dual-ESI source operated in the negative ion mode in soft conditions [48 (link)]. The following parameters were used: fragmentor 320 V, trap fill time 1000 µs, trap release time 200 µs, trap entrance grid delta 2 V. The drift tube was filled with helium and the pressure was fixed at 3.89 ± 0.01 Torr (measured by a capacitance diaphragm gauge CDG-500, Agilent Technologies).
The MS data were extracted using the IM-MS Browser (v. B.08.00; Agilent Technologies). The peak area intensity of each species at the 5- charge state was extracted from their corresponding m/z range (encompassing potassium adducts), and baseline subtracted. The concentrations of all species (free DNA, free ligands, and complexes) and corresponding dissociation constants were determined as previously described [49 (link),50 (link)].
The MS data were extracted using the IM-MS Browser (v. B.08.00; Agilent Technologies). The peak area intensity of each species at the 5- charge state was extracted from their corresponding m/z range (encompassing potassium adducts), and baseline subtracted. The concentrations of all species (free DNA, free ligands, and complexes) and corresponding dissociation constants were determined as previously described [49 (link),50 (link)].
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