Jet stream esi source

Manufactured by Agilent Technologies

Sourced in United States, Germany

The Jet Stream ESI source is a core component of Agilent's mass spectrometry instruments. It is designed to efficiently introduce liquid samples into the mass spectrometer for analysis. The Jet Stream technology employs a unique spray geometry to optimize the ionization process, enabling improved sensitivity and robustness across a wide range of applications.

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16 protocols using jet stream esi source

HPLC-MS/MS Quantification of 6dEB

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All experiments were carried out on an Agilent 1200 HPLC system coupled to a 6460 Triple Quadrupole mass spectrometer equipped with a Jet stream ESI-source (Agilent Technologies, Waldbronn, Germany). The chromatographic separation was achieved temperature controlled at 25 °C on a Synergi Fusion-RP column (2.5 μm, 50 × 2.0 mm) equipped with a pre-column of the same material (4 × 2.0 mm) both from Phenomenex (Aschaffenburg, Germany). A gradient of mobile phase A (0.1 % formic acid) and mobile phase B (acetonitrile) was used as shown in Table S2 in the Supplementary Material (flow rate of 0.5 mL/min, injection volume of 10 μL). The MS was operated in positive ion mode with multi-reaction monitoring (MRM). The MS/MS fragmentation pattern of 6dEB and the internal standard compounds were determined and the MS parameters were optimized. The optimized source parameters are displayed in Table S3 in the Supplementary Material.
6dEB was monitored with transitions m/z 409.1 to m/z 311.2 for relative quantification and m/z 409.1 to m/z 391.2 and m/z 293.2 for identification. Collision energies for the transitions of 6dEB were 25, 21, and 25 V, respectively. Data was acquired and evaluated using the Mass Hunter software; quantitative analysis was done using Mass Hunter Quantitative Analysis (version B03.02, Agilent Technologies, Waldbronn, Germany).

Kumpfmüller J., Methling K., Fang L., Pfeifer B.A., Lalk M, & Schweder T. (2015). Production of the polyketide 6-deoxyerythronolide B in the heterologous host Bacillus subtilis. Applied Microbiology and Biotechnology, 100, 1209-1220.

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HPLC-QTOF-MS Analysis of Compounds

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An Agilent 1100 HPLC equipped with DAD and FLD was employed in the preliminary studies; the LC-MS analyses were performed on a 1290 UHPLC coupled to a 6550 iFunnel QTOF MS with a Jet Stream ESI source (Agilent, Santa Clara, CA, USA). The mobile phase gradient and flow rates are depicted in Table 9.
An Agilent Dual Jet Stream electrospray (ESI) source was used in positive ionization mode with nitrogen as drying, nebulizer and sheath gas. Parameters such as the sheath gas temperature, ESI source and nozzle voltages, TOF fragmentor voltage, collision energies and mass acquisition rates were optimized to achieve the best analyte responses in the desired concentration ranges. The following final conditions were applied: drying gas temperature 200 °C; drying gas flow 14 L/min; nebulizer pressure 35 psi; sheath gas flow 11 L/min. TOF mass calibration was performed daily in the extended dynamic range (2 GHz) and low mass range (1700 m/z). The reference masses used for the within-run mass correction were m/z 121.0509 and 922.0098.
Data were collected using an Agilent MassHunter Workstation software Qualitative analysis 10.0 and Data Acquisition for 6200 series TOF/6500 series QTOF 10.1 (Santa Clara, CA, USA). Microsoft Office 365 Excel (Redmond, WA, USA) with the PK solver extension and GraphPad Prism 8 (San Diego, CA, USA) were used for data analysis.

Turković L., Bočkor L., Ekpenyong O., Silovski T., Lovrić M., Crnković S., Nigović B, & Sertić M. (2022). Development and Validation of a Novel LC-MS/MS Method for the Simultaneous Determination of Abemaciclib, Palbociclib, Ribociclib, Anastrozole, Letrozole, and Fulvestrant in Plasma Samples: A Prerequisite for Personalized Breast Cancer Treatment. Pharmaceuticals, 15(5), 614.

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Collision Cross-Section Analysis of Anabolic Steroids

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AASs were analyzed in negative ionization mode using the Jet Stream ESI source (Agilent) coupled with a drift tube ion mobility mass spectrometer (6560, Agilent) using settings similar to previously described instrumental methods.^{38 (link),44 (link),46 (link),47 (link),50 (link),78 (link)} Ionization source conditions were optimized (e.g., gas temperature, drying gas, nebulizer pressure, sheath gas temperature, sheath gas flow, capillary voltage, and nozzle voltage in Table S1) for flow injection analysis (FIA) to maximize sensitivity. The IM analyses used nitrogen drift gas with the drift tube at a temperature of 30 °C, a pressure of 4.0 Torr, and an electric field of 17.3 V/cm. A single field CCS method was used to determine CCS values via a modified Mason-Schamp equation.^{46 (link)} Data were acquired using MassHunter Workstation Data Acquisition software (Agilent) and analyzed using MassHunter Qualitative Analysis (Agilent), MassHunter IM-MS Browser (Agilent), and Skyline (MacCoss Lab).^{79 (link),80 (link)} Statistical analyses for isomeric AAS phase II metabolite RT and CCS measurements were performed using GraphPad Prism (version 8.0). Significant difference was assessed based on a p-value < 0.05 from appropriate statistical tests (t-tests (Tables S2–S3) and one-way analysis of variance (ANOVA) tests (Tables S4–S5)).

Davis DE J.r., Leaptrot K.L., Koomen D.C., May J.C., Cavalcanti G.D., Padilha M.C., Pereira H.M, & McLean J.A. (2021). Multidimensional Separations of Intact Phase II Steroid Metabolites Utilizing LC-Ion Mobility-MS. Analytical chemistry, 93(31), 10990-10998.

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Accurate Step-Field Collision Cross-Section Measurements

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Samples were infused at 15 μL/min via syringe pump into an Agilent Jetstream ESI source operated in either positive or negative mode. Accurate step-field ^DTCCS_XX measurements (where the subscript ‘XX’ indicates the specific gas used for each measurement) were performed using the step-field method with five field strength steps (30 s each) ranging from 13.5 to 18.6 V/cm for all gases except helium, for which the field range was lowered to 9.6–14.7 V/cm to prevent discharge in the drift tube. The effective drift tube length was measured using the method from the McLean group’s Collision Cross Section Compendium Reporting Guidelines; this value was used to adjust experimentally obtained ^DTCCS_XX values. Triplicate measurements were made in all cases. Single-field ^DTCCS_XX measurements [33] (link) were performed following chromatographic separation at a drift tube field strength of 18.6 V/cm for all gases except helium, for which measurements were made at 14.7 V/cm. The Agilent Tune Mix reference standard was used to create a calibration line with slope (beta) and intercept (tfix) to convert drift time to ^DTCCS_XX. Reference CCS values for the Tune Mix ions in He, Ar, and CO₂ were found in work by Morris et al.[34] (link).

Neal S.P., Wilson K.M., Velosa D.C, & Chouinard C.D. (2022). Targeted glucocorticoid analysis using ion mobility-mass spectrometry (IM-MS). Journal of Mass Spectrometry and Advances in the Clinical Lab, 24, 50-56.

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Characterization of SRAPd-Abasic DNA Complex

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SRAPd was incubated with the abasic site containing 3’ overhang DNA (same as used in SRAPd_DPC co-crystallization) at 1:1.2 ratio in a buffer containing 150 mM NaCl, 20 mM Hepes pH 7.5, 10 mM MgCl₂ at room temperature overnight. All LC-MS data were acquired according to the previously published protocol²³, on an Agilent 6545 Q-TOF (Santa Clara, CA) equipped with a Dual Agilent Jet Stream ESI source coupled with an Agilent 1260 Infinity HPLC system (Santa Clara, CA). The analytical column utilized was a 300 StableBond Poroshell (Agilent, part number 883750–909) 2.1 × 100-mm-i.d. reversed-phase C₃ (5 μm particle size). Mobile phase (A) consisted of 97% HPLC grade water with 0.5% formic acid and 2.5% ACN, while mobile phase (B) was 96% ACN with 0.5% formic acid and 3.5% HPLC grade water. A gradient profile was utilized at a flow rate of 500 μL/min. The mobile phase was held for 2 min at 5% B (with eluant going to waste) and then switched to the mass spectrometer from 2–6 min during which time solvent B increased from 5–95%. Two microliters of a 30μM solution of each sample was injected. Raw data files were analyzed by Agilent MassHunter BioConfirm software (vB.07.00). Mass spectra between 4 and 6 minutes were extracted, averaged and deconvoluted using the MaxEnt algorithm.

Halabelian L., Ravichandran M., Li Y., Zeng H., Rao A., Aravind L, & Arrowsmith C.H. (2019). Structural basis of HMCES interactions with abasic DNA and multivalent substrate recognition. Nature structural & molecular biology, 26(7), 607-612.

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Characterization of Metabolite Extracts

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The constitution of ME was evaluated by ultra-high performance LC, using a Zorbax Eclipse Plus C18 Rapid Resolution HD column (2.1 × 150 mm, 1.8 μm) and an LC-Q-TOF-MS apparatus to detect positive and negative electrospray ionization (ESI) mass spectra. The temperature was maintained at 40 °C. Mobile phase A comprised 0.1% formic acid in water, and mobile phase B comprised 11.5% acetonitrile solution. The elution steps were as follows [1 ]: 70% A and 30% B for 2 min [2 ], 5% A and 95% B for 14 min [3 ], 70% A and 30% B for 14.20 min, and [4 ] 70% A and 30% B for 20 min. The sample volume was 2 μL, with a flow rate of 0.2 mL/min. The drying gas temperature was set to 350 °C at 10 L/min; sheath gas temperature, to 275 °C; and nebulizer pressure, to 60 psig. Using a dual Agilent Jet Stream ESI source, we conducted LC–MS/MS in positive and negative ion modes, with a scanning range of 100–1500 m/z.

Pooprommin P., Manaspon C., Dwivedi A., Mazumder A., Sangkaew S., Wanmasae S., Tangpong J., Ongtanasup T, & Eawsakul K. (2022). Alginate/pectin dressing with niosomal mangosteen extract for enhanced wound healing: evaluating skin irritation by structure-activity relationship. Heliyon, 8(12), e12032.

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Quantitative Analysis of Fructoselysine

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The determination of fructoselysine was carried out on An Agilent 1290 UPLC system combined with an Agilent 6460 triple-quadrupole mass spectrometry equipped with a Jet Stream ESI source (Agilent Technologies Inc. Santa Clara, CA, USA). A Phenomenex Synergi Hydro-RP column (250 mm × 2 mm, 4 μm, 80 Å; Phenomenex, Torrance, CA, USA) was used for the chromatographic separation. The binary mobile phase compositions were methanol and 0.1% (v/v) formic acid in water (30 : 70, v/v). The isocratic condition was applied at a flow rate of 0.2 mL min⁻¹. The temperature of the column oven was maintained at 25 °C and the injection volume used was set at 5 μL.
The identification and quantitation of fructoselysine was achieved using the positive ESI and MRM mode. The optimized parameters used for this analysis are presented in the ESI (Table S1†). The quantitation and qualification ions of fructoselysine were m/z 309 → 84 and m/z 309 → 147, respectively. The optimal settings of MS for fructoselysine detection are reported in the ESI (Table S2†). The total and extracted ion chromatogram of each specific transition is also shown in the ESI (Fig. S1†). The fructoselysine was calibrated by a standard curve (r² = 0.9990) built with fructoselysine standard gradients.

Wang Y., Hu H., McClements D.J., Nie S., Shen M., Li C., Huang Y., Chen J., Zeng M, & Xie M. (2019). Effect of fatty acids and triglycerides on the formation of lysine-derived advanced glycation end-products in model systems exposed to frying temperature. RSC Advances, 9(27), 15162-15170.

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Denaturing LC-MS Analysis of mCP and mS100 Proteins

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A denaturing protocol was employed to analyze mCP, mCP-Ser, mS100A8 and mS100A9 by LC-MS. An Agilent 1260 series LC system outfitted with an Agilent Jetstream ESI source and an Agilent Poroshell 300SB-C18 column (5-μm pore size) was used for all analyses. Each protein was diluted in Milli-Q water to provide a final concentration of ≈5 μM. A 5-μL protein sample was injected onto the column, and the mS100A8 and mS100A9 subunits were eluted using a gradient of 10–90% B over 20 min with a flow rate of 0.2 mL/min (solvent A: 0.1% formic acid in water; solvent B: 0.1% formic acid in acetonitrile). The spectra were deconvoluted using the maximum entropy algorithm in the MassHunter software (Agilent).

Hadley R.C., Gu Y, & Nolan E.M. (2018). Initial Biochemical and Functional Evaluation of Murine Calprotectin Reveals Ca(II)-Dependence and Its Ability to Chelate Multiple Nutrient Transition Metal Ions. Biochemistry, 57(19), 2846-2856.

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Quantitative Analysis of Eicosanoids

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The samples were analyzed using a Zorbax Eclipse XDB-C18 column (5 µm, 4.6 × 150 mm), run on the Agilent 1200 Series HPLC system (Santa Clara, CA, USA), connected to a diode array detector, followed by the 500TR Series Flow Scintillation Analyzer (Packard Bioscience, Meriden, CT, USA) or the Agilent 6540 UHD Accurate Quadrupole Time-of-Flight MS/MS with an Agilent Jet Stream™ ESI source in negative mode(Santa Clara, CA, USA). The elution was performed with a solvent system of ACN/water/formic acid (98.9%/1.0%/0.1% v/v/v) (A) and water/formic acid (99.9%/0.1% v/v) (B), 0–8 min isocratic (35%A:65%B), 9–17 min gradient to 100% A, 18–30 min 100% A at a flow rate of 1 mL min⁻¹ [26 (link)]. The data were analyzed by Agilent MassHunter Workstation Software Qualitative Analysis, Version B.05.00 Build 5.0.519.0. Eicosanoids were identified by comparing their retention times and mass spectra with those of authentic standards.

Lõhelaid H, & Samel N. (2018). Eicosanoid Diversity of Stony Corals. Marine Drugs, 16(1), 10.

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Agilent LC-MS/MS method for tetrodotoxin

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An Agilent 1290 Infinity LC system (Agilent Technologies Deutschland GmbH, Waldbronn, Germany) was used for the liquid chromatographic separation. This separation is described in Leão et al. [18 (link)]. The chromatographic conditions used in the analysis of TTX are summarized in Table S6 (Supplementary Materials).
A 6460A Triple Quadrupole mass/massMS/MS (QQQ) equipped with a Jet Stream ESI source (Agilent Technologies Deutschland GmbH Waldbronn, Germany) was used for the analysis of TTX in MRM (multiple reaction monitoring) mode by detecting m/z transitions in tandem mass spectrometry, which are included in Table S7 of Supplementary Materials [20 ].
The optimized conditions are summarized in Table S8 of Supplementary Materials [18 (link)].

López Cabo M., Romalde J.L., Simal-Gandara J., Gago Martínez A., Giráldez Fernández J., Bernárdez Costas M., Pascual del Hierro S., Pousa Ortega Á., Manaia C.M., Abreu Silva J, & Rodríguez Herrera J. (2020). Identification of Emerging Hazards in Mussels by the Galician Emerging Food Safety Risks Network (RISEGAL). A First Approach. Foods, 9(11), 1641.

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