Hesi probe

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany

The HESI probe is a core component of mass spectrometry instrumentation, primarily used for the ionization of analytes prior to detection. It utilizes the heated electrospray ionization (HESI) technique to generate ions from liquid samples, enabling efficient and sensitive analysis of a wide range of compounds.

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12 protocols using hesi probe

Endophyte Isolation and Quantification

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Mass spectrometry (MS) analysis was carried out on a TSQ Quantum Access MAX triple stage quadrupole mass spectrometer with a heated electrospray ionization (HESI) probe (Thermo Scientific, San Jose, CA). The MS conditions were as follows: ion source polarity: negative or positive; spray voltage: −2200 V or 2800 V; vaporizer temperature: 300°C; skimmer offset: 3 V; sheath gas pressure (N₂): 30 (arbitrary units); auxiliary gas pressure (N₂): 5 (arbitrary units); collision pressure: 1.2 m Torr; ion transfer tube temperature: 300°C; scan type: selective reaction monitoring.
In order to elucidate the exact contribution of endophyte on the effect of phytohormones in plants, the growth dynamics of inoculated bacteria were investigated. Therefore, endophytic bacteria were simultaneously isolated from the roots, stems and leaves of the inoculated tissue culture seedlings of D. officinale every other week. After washing the seedlings using sterile tap water, 1 g each of root, stem and leaf samples was pulverized and diluted in dd H₂O, and spread on the NA medium. The resulting bacterial populations were calculated using the plate counting method.

Yang S., Zhang X., Cao Z., Zhao K., Wang S., Chen M, & Hu X. (2014). Growth-promoting Sphingomonas paucimobilis ZJSH1 associated with Dendrobium officinale through phytohormone production and nitrogen fixation. Microbial Biotechnology, 7(6), 611-620.

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Quantitative Analysis of Cefotaxime

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Cefotaxime concentrations in plasma were quantified with a UPLC–MS/MS system consisting of a Dionex Ultimate UPLC system connected to a triple quadrupole mass spectrometer (Thermo TSQ Vantage with HESI-probe, Thermo Scientific) [10 ]. The lower and upper levels of quantification of this method were 0.13 and 12.5 mg/l, respectively.

Hartman S.J., Boeddha N.P., Ekinci E., Koch B.C., Donders R., Hazelzet J.A., Driessen G.J, & de Wildt S.N. (2019). Target attainment of cefotaxime in critically ill children with meningococcal septic shock as a model for cefotaxime dosing in severe pediatric sepsis. European Journal of Clinical Microbiology & Infectious Diseases, 38(7), 1255-1260.

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Negative Ionization Mass Spectrometry

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After InsPs extraction with TiO₂, a total of 150 μL of the reconstituted eluate was separated, and 10 μL was injected into a Q Exactive Orbitrap high-resolution mass spectrometer that had a heated electrospray ionization (HESI) probe (Thermo Fisher Scientific, Waltham, USA), which was operated in negative ionization mode. An example of Mass spectrum is provided in Fig 1.

Grases F., Costa-Bauzá A., Berga F., Gomila R.M., Martorell G, & Martínez-Cignoni M.R. (2019). Intake of myo-inositol hexaphosphate and urinary excretion of inositol phosphates in Wistar rats: Gavage vs. oral administration with sugar. PLoS ONE, 14(10), e0223959.

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Quantitative HILIC-MS Metabolite Analysis

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Scan type: full MS in the positive-and-negative mode (alternating); scan range, 69–1000 m/z; resolution, 70,000; AGC-target, 3E6; maximum injection time, 200 ms; sheath gas, 30; auxiliary gas, 10; sweep gas, 3; spray voltage, 3.6 kV (positive mode) or 2.5 kV (negative mode); capillary temperature, 320 °C; S-lens RF level, 55.0; and auxiliary gas heater temperature, 120 °C. Annotation and data evaluation: Peaks corresponding to the calculated monoisotopic masses (MIM ± H⁺ ± 2 mMU) were integrated using TraceFinder software (Thermo Scientific, Bremen, Germany). Materials: Ultrapure water was obtained from a Millipore water purification system (Milli-Q Merck Millipore, Darmstadt, Germany). HPLC–MS solvents, LC–MS NH₄OAc and lamivudine were purchased from Merck. RP18-SPE Columns: 50 mg Strata C18-E (55 µm) in 1 mL tubes (Phenomenex, Aschaffenburg, Germany). Sonifier: Branson Ultrasonics 250 equipped with a 13 mm-Disintegrator-Sonotrode (Thermo Scientific). LC/MS-system: Thermo Scientific Dionex UltiMate 3000 UHPLC system hyphenated with a Q Exactive mass spectrometer (QE-MS) equipped with a HESI probe (Thermo Scientific). Particle Filter: Javelin Filter with an ID of 2.1 mm (Thermo Scientific). UPLC-precolumn: SeQuant ZIC-HILIC column (5 μm particles, 20 × 2 mm) (Merck). UPLC column: SeQuant ZIC-HILIC column (3.5 μm particles, 100 × 2.1 mm) (Merck).

Schmitz W., Koderer C., El-Mesery M., Gubik S., Sampers R., Straub A., Kübler A.C, & Seher A. (2021). Metabolic Fingerprinting of Murine L929 Fibroblasts as a Cell-Based Tumour Suppressor Model System for Methionine Restriction. International Journal of Molecular Sciences, 22(6), 3039.

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Comprehensive Lipid Profiling by LC-MS/MS

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A Thermo Vanquish UPLC system coupled with Q-Exactive Orbitrap mass spectrometer equipped with a heated electrospray ionization (HESI) probe (Thermo Fisher, CA, USA) was used in this study. An Acquity UPLC CSH C18 1.7μm 2.1×100mm column (Waters Corp, Milford, MA, USA) was applied for lipid separation in both negative and positive ionization modes with separate injections. Mobile phase A was 100% water, containing 5 mM ammonium acetate and 0.1% formic acid, and mobile phase B was acetonitrile and H₂O at a proportion of 95/5 (v/v), containing 5 mM ammonium acetate and 0.1% formic acid. A linear gradient elution program was set as 35% B from 0–1 min, increasing to 95% B over 20 min; after another 10 min at 95% B, the mobile phase composition was then returned to 35% B. The total run time was 35 min. The flow rate was 0.4 mL/min and the column temperature was 40°C. The MS/MS data were acquired at the ranges of m/z 100–1500. The resolution for data collection in full scan and fragment spectra were 140000 and 17500, respectively. AGC target was 3e⁶ and the maximum IT was 50ms for full scan mode. While the MS/MS mode parameters were set as automatic gain control (AGC) target at 1e⁵, maximum IT of 50ms, loop count at 10, isolation window of 1.2 m/z, and normalized collision energy (NCE) of 20, 50, and 80 eV.

Chen L., Zhang S., Sun X., McDonald J.D., Bruno R.S, & Zhu J. (2021). Application of comparative lipidomics to elucidate postprandial metabolic excursions following dairy milk ingestion in individuals with prediabetes. Journal of proteome research, 20(5), 2583-2595.

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UPLC-MS/MS Analytical Protocol

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A Dionex Ultimate UPLC system consisting of an Ultimate 3,000 RS UPLC pump, an Ultimate 3,000 RS autosampler and an Ultimate 3,000 RS Column Compartment was used as the equipment. The UPLC was connected to a Thermo TSQ Vantage triple quadrupole MS with HESI probe (Thermo Scientific, Waltman, MA, USA). The software programs Chromeleon (version 6.8, Dionex, Thermo Scientific), Xcalibur (version 2.1, Thermo Scientific) and LCquan (version 2.6, Thermo Scientific) were used to control the system and analyze the data.

Flint R.B., Bahmany S., van der Nagel B.C, & Koch B.C. (2018). Simultaneous quantification of fentanyl, sufentanil, cefazolin, doxapram and keto‐doxapram in plasma using liquid chromatography–tandem mass spectrometry. Biomedical Chromatography, 32(10), e4290.

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High-Resolution Mass Spectrometry Workflow

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Diluted samples (1:1000) were injected into a Q Exactive Orbitrap high-resolution mass spectrometer equipped with a heated electrospray ionization (HESI) probe (Thermo Fisher Scientific, Waltham, USA), which was operated in a negative ionization mode. The temperature of the ion transfer capillary was set to 320 °C, the spray voltage was set to 2.9 kV in negative mode and the S-lens RF level was 50 AU. Direct injection in the full scan acquisition mode over a range of 150 to 700 m/z was performed with a resolution of 140,000.

Grases F., Costa-Bauzá A., Calvó P., Julià F., Dietrich J., Gomila R.M., Martorell G, & Sanchis P. (2022). Phytate Dephosphorylation Products Also Act as Potent Inhibitors of Calcium Oxalate Crystallization. Molecules, 27(17), 5463.

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LC-MS Analysis of Unknown Compounds

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LC/MS analysis was performed using an Accela UHPLC system (Thermo Fisher Scientific, Bremen, Germany) coupled to an LTQ OrbiTrap mass spectrometer equipped with a heated electrospray ionization (HESI) probe (Thermo Fisher Scientific, Bremen, Germany). A Syncronis C18 column (100 × 2.1 mm, 1.7 µm particle size) from Thermo Fisher Scientific was used as the analytical column for separation. The chromatographic and MS settings were the same as previously described in the literature [5 (link)]. Tentative identification of unknown compounds was conducted based on their monoisotopic mass and MS³ fragmentation, and confirmed using previously reported MS fragmentation or data [6 (link),7 (link),8 (link),9 (link),10 (link),11 (link)]. ChemDraw software (version 12.0, CambridgeSoft, Cambridge, MA, USA) was used to calculate the accurate mass of compounds of interest. Xcalibur software (version 2.1, Thermo Fisher Scientific, Waltham, MA, USA) was used for instrument control, data acquisition, and data analysis.

Milutinović M., Nakarada Đ., Božunović J., Todorović M., Gašić U., Živković S., Skorić M., Ivković Đ., Savić J., Devrnja N., Aničić N., Banjanac T., Mojović M, & Mišić D. (2023). Solanum dulcamara L. Berries: A Convenient Model System to Study Redox Processes in Relation to Fruit Ripening. Antioxidants, 12(2), 346.

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Enzymatic Analysis of Porcine Pancreas α-Amylase

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All chemicals were obtained from commercial suppliers and used without further purification. ¹H (600 MHz) and ¹³C (deduced from HSQC and HMBC spectra) NMR spectra were recorded using a Bruker AVANCE III, with CDCl₃ as the solvent and TMS as the internal standard. Melting points were measured using a Buchi melting point apparatus, model B-545. ESI-HRMS spectra were recorded with a platform consisting of an HTC PAL^® 4 autosampler (CTC Analytics AG, Zwingen, Switzerland), an Accela U-HPLC system with quaternary pumps, and an Exactive Orbitrap mass spectrometer equipped with a heated electrospray ionization (H-ESI) probe (both from Thermo Fisher Scientific, Bremen, Germany). α-Amylase was extracted from porcine pancreas (Sigma Aldrich 6255, 1151 Umg⁻¹ of protein, Sigma Aldrich, Saint Louis, MO).

Baccari W., Saidi I., Filali I., Znati M., Lazrag H., Tounsi M., Marchal A., Waffo-Teguo P, & Ben Jannet H. (2024). Semi-synthesis, α-amylase inhibition, and kinetic and molecular docking studies of arylidene-based sesquiterpene coumarins isolated from Ferula tunetana Pomel ex Batt. RSC Advances, 14(7), 4654-4665.

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UHPLC-MS Analysis Workflow

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The equipment used for LC/MS analysis was a Thermo Scientific Dionex Ultimate 3000 UHPLC system hyphenated with a Q Exactive Mass Spectrometer (QE-MS) equipped with a heated electrospray ionization (HESI) probe (Thermo Scientific, Bremen, Germany).

Bohnert S., Reinert C., Trella S., Schmitz W., Ondruschka B, & Bohnert M. (2020). Metabolomics in postmortem cerebrospinal fluid diagnostics: a state-of-the-art method to interpret central nervous system–related pathological processes. International Journal of Legal Medicine, 135(1), 183-191.

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