Isq single quadrupole mass spectrometer

Manufactured by Thermo Fisher Scientific

Sourced in United States, Austria

The ISQ single quadrupole mass spectrometer is a laboratory instrument designed for qualitative and quantitative analysis of chemical compounds. It uses a quadrupole mass analyzer to separate and detect ions based on their mass-to-charge ratio. The core function of the ISQ single quadrupole mass spectrometer is to provide accurate mass measurements and identification of sample components.

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37 protocols using isq single quadrupole mass spectrometer

Fatty Acid Profiling by GC-MS

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The recovered fatty acid methyl esters were chromatographically analyzed using GC-MS [76 (link)]. TRACE^® GC Ultra Gas Chromatograph (Thermo Scientific Corp., Berkeley, MO, USA) was used in conjunction with a Thermo MS detector (ISQ^® Single Quadrupole Mass Spectrometer, Thermo Fisher Scientific, Berkeley, MO, USA). The system included a TR-5 MS column (30 m × 0.32 mm i.d., 0.25 m film thickness).
The system was set up to analyze 1 L diluted samples (1:10 hexane, v/v), helium as the carrier gas, and the injector and detector at 210 °C. The flow rate was set to 1.0 mL/min with a split ratio of 1:10. The temperature program was 60 °C for 1 min, then rose at 4.0 °C/min to 240 °C for 1 min. Electron ionization (EI) at 70 eV yielded mass spectra with a spectral range of m/z 40–450. Finally, the obtained MS data were de-convoluted using AMDIS software (www.amdis.net, accessed on 20 October 2021) and identified by retention indices (relative to n-alkanes C8-C22), mass spectrum matching to authentic standards (when available), and Wiley spectral library collection and NIST library database.

Elmaidomy A.H., Mohamed E.M., Aly H.F., Younis E.A., Shams S.G., Altemani F.H., Alzubaidi M.A., Almaghrabi M., Harbi A.A., Alsenani F., Sayed A.M, & Abdelmohsen U.R. (2022). Anti-Inflammatory and Antioxidant Properties of Malapterurus electricus Skin Fish Methanolic Extract in Arthritic Rats: Therapeutic and Protective Effects. Marine Drugs, 20(10), 639.

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Soil Microbial Community Profiling by PLFA

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Soil microbial community structure was determined by PLFA profiling. PLFAs were extracted according to Kaiser et al. (2010b) (link) and analyzed with gas chromatography (GC) (Trace 1300, Thermo Scientific, Austria) coupled to an ISQ single quadrupole mass spectrometer (Thermo Scientific, Germany). We used branched PLFAs (a15:0, i15:0, i16:0, a17:0, and i17:0) as indicator for gram-positive bacteria, cyclopropyl and mono-unsaturated PLFAs (16:1ω7, 16:1ω9, cy17:0, and cy19:0) for gram-negative bacteria, saturated PLFAs (15:0 and 17:0) for uncategorized bacteria, poly-unsatured PLFAs (18:1ω9 and 18:2ω6,9) for fungi, 10Me-PLFAs (10Me17:0, 10Me18:0, 10Me19:0) for actinomycetes, and unspecific PLFAs (14:0, 16:0,18:0, 20:0) for viable biomass (Hill et al., 2000 ; Kaiser et al., 2010a (link); Inglett et al., 2011 ). Fungi to bacteria ratios (F/B) were calculated by dividing fungal PLFAs by the sum of bacterial PLFAs. The fungal biomarker PLFA 18:1w9 however also occurs in some gram-positive bacteria, eventually causing an overestimation of fungal biomass and fungi to bacteria ratios in agricultural soils (Frostegard et al., 2011 ).

Hu Y., Zheng Q., Zhang S., Noll L, & Wanek W. (2018). Significant release and microbial utilization of amino sugars and D-amino acid enantiomers from microbial cell wall decomposition in soils. Soil biology & biochemistry, 123, 115-125.

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Analytical Characterization of Organic Compounds

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Mass spectra were measured with Thermo scientific, ISQ single quadrupole mass spectrometer (San Jose, CA, USA). NMR analysis was measured on ¹H-NMR (300 MHz), ¹³C-NMR (75 MHz): Varian Mercury-VX-300 spectrophotometer. Thin layer chromatography (TLC) was performed on precoated silica gel plates using solvent systems S₁: n-c₆H₁₄: Ethyl acetate (EtOAc) (95:5), S₂: N-C₆H₆: EtOAc (9:1), and S₃: n-c₆H₁₄: EtOAc (8:2), the spots were detected by spraying with p-anisaldehyde-sulfuric acid spray reagent.

Abdel-Monem A.R, & Abdelrahman E.H. (2016). New Abietane Diterpenes from Euphorbia Pseudocactus Berger (Euphorbiaceae) and Their Antimicrobial Activity. Pharmacognosy Magazine, 12(Suppl 3), S346-S349.

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Gas Chromatography-Mass Spectrometry Analysis

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The non-polar extract of the fungal mycelium was analyzed with a TRACE GC Ultra Gas Chromatograph instrument (Thermo Fisher Scientific, Waltham, MA, USA) equipped with both a thermal mass spectrometer detector (ISQ Single Quadrupole Mass Spectrometer) and a TR-5 MS column (30 m × 0.32 mm i.d., 0.25 µm film thickness). The experiment was conducted with helium serving as the carrier gas at a flow rate of 1.0 mL/min and a split ratio of 1:10. After setting it at 60 °C for one min, the temperature was ramped at a rate of 4 °C/min to 240 °C and then maintained for one minute. Both the injector and the detector were kept at a temperature of 210 °C. The sample was diluted 1:10 with hexane (v/v before being injected as 1.0 µL aliquot. The mass spectral range was set to m/z 40–450; electron ionization (EI) was performed at 70 eV. The identification of the non-polar metabolites was performed using the AMDIS program (www.amdis.net, accessed on 23 October 2022), retention indices (relative to n-alkanes C8-C22), and mass spectrum, which were matched to authentic standards (once it becomes accessible) as well as NIST databases and Wiley spectral library.

El-Sayed H., Osman M.E., Abdelsalam A., Boroujerdi A., Sonbol H, & Elsaba Y.M. (2022). Morphological, Molecular and Metabolic Characterization of the Pigmented Fungus Subramaniula asteroids. Journal of Fungi, 8(11), 1149.

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Gas Chromatographic Analysis of Fermentation Samples

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Fermentation samples with a volume of 2 ml were analyzed in 15-ml vials with a Thermoscience Trace gas chromatograph equipped with a ResTek Stabilowax polyethylene glycol column with a 0.25-µm diameter. Split injection (1:25), with a resting flow rate of 2 ml/min of He, was performed after 10 min of equilibration at 60°C with a Thermoscience TriPlus RSH autosampler. The oven was kept at 40°C for 2 min, heated to 240°C with a ramping of 15°C/min, and kept at 240°C for 2 min. The detection was carried out with a flame ionization detector (FID), using 20 ml/min of N₂, 350 ml/min high-grade compressed air, and 30 ml/min H₂ provided from a VWR H₂ generator. For the industrial strains, quantification was done under identical chromatography conditions, except that detection was performed with a Thermo Scientific ISQ single quadrupole mass spectrometer. Concentrated standards were kept at −20°C in gas chromatography (GC)-grade absolute ethanol and brought to the final concentration in 5% ethanol in volumetric dilutions, except for wine and saké fermentations, for which 15% ethanol was used in the standards. Fermentation samples were kept at 4°C prior to analysis.

Holt S., Trindade de Carvalho B., Foulquié-Moreno M.R, & Thevelein J.M. (2018). Polygenic Analysis in Absence of Major Effector ATF1 Unveils Novel Components in Yeast Flavor Ester Biosynthesis. mBio, 9(4), e01279-18.

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GC-MS Characterization of Milk Fats and Oils

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The GC-MS analysis of cow, camel, goat, and human milk fats, as well as rice bran, canola, and coconut oils, was carried out using gas chromatography–mass spectrometry instrument stands with the following specifications: a TRACE GC Ultra Gas Chromatographs (THERMO Scientific Corp., Waltham, MA, USA), coupled with a thermal mass spectrometer detector (ISQ Single Quadrupole Mass Spectrometer). The GC-MS system was equipped with a TR-5 MS column (30 m × 0.32 mm i.d., 0.25 μm film thickness). Analyses were carried out using helium as carrier gas at a flow rate of 1.0 mL/min and a split ratio of 1:10 using the following temperature program: 60 °C for 1 min, rising at 4.0 °C/min to 240 °C and held for 1 min. The injector and detector were held at 210 °C. Diluted samples (1:10 hexane, v/v) of 1μL of the mixtures were continuously injected. Mass spectra were obtained by electron ionization (EI) at 70 eV, using a spectral range of m/z 40–450. The identification of the chemical constituents of the essential oil was de-convoluted using AMDIS software “www.amdis.net (accessed on 2 August 2022)” by its retention indices (relative to n-alkanes C4–C22) and mass spectrum matching to authentic standards (when available), Wiley spectral library collection, and NSIT library database.

Al-Nassir N.S, & Sakr S.S. (2024). In Vitro Digestibility Assessment of Whey from Goat and Camel Milk Fermented with Lactobacillus helveticus for Use as a Base in Formulating Follow-On Formula. Foods, 13(4), 570.

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Metabolite Derivatization and GC-MS Analysis

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The dried samples, QC sample, and processing blank sample, were derivatized using methoxyamine hydrochloride (MOX) and N,O-Bis(trimethylsilyl)trifluoroacetamide (TMS) (Sigma). Briefly, dried extracts were reconstituted in 30 μL of 11.4 mg/ml MOX in anhydrous pyridine (VWR), vortexed for 10 minutes, and heated at 60°C for 1 hour. Next, 20 μL TMS was added to each reconstituted extract, vortexed for 1 minute, and heated at 60°C for 30 min. The derivatized samples, QC samples and processing blank samples were immediately analyzed using GC/MS.
GC chromatographic separation was conducted on a Thermo Trace 1300 GC with a TraceGold TG-5SilMS column (0.25 μM film thickness; 0.25mm ID; 30 m length). 1 μL of derivatized sample, QC, or blank was injected. The GC was operated in split mode with the following settings: 20:1 split ratio; split flow: 24 μL/min, purge flow: 5 mL/min, Carrier mode: Constant Flow, Carrier flow rate: 1.2 mL/min). The GC inlet temperature was 250°C. The GC oven temperature gradient was as follows: 80°C for 3 min, ramped at 20°C/min to a maximum temperature of 280°C, which was held for 8 min. The injection syringe was washed 3 times with pyridine between each sample. Metabolites were detected using a Thermo ISQ single quadrupole mass spectrometer. The data was acquired from 3.90 to 21.00 min in EI mode (70eV) by single ion monitoring (SIM).

Buchanan J.L., Rauckhorst A.J, & Taylor E.B. (2023). 3-hydroxykynurenine is a ROS-inducing cytotoxic tryptophan metabolite that disrupts the TCA cycle. bioRxiv.

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Synthesis and Characterization of Novel Compounds

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All of the corrected melting points are in degree centigrade and measured using a Stuart SMP20 melting point apparatus (Bibby Scientific Limited, Staffordshire, UK). The infrared spectra were recorded on a PerkinElmer Alpha platinum-ATR spectrometer; the ¹H-NMR (300 MHz) and ¹³C-NMR (75 MHz) spectra were recorded on a Varian Mercury VXR-300 spectrometer (Varian Inc., Palo Alto, CA, USA) and the chemical shifts were related to that of the solvent DMSO-d₆ using tetramethylsilane (TMS) as an internal standard. MS spectra (HRMS) were acquired on TRACE GC Ultra gas chromatograph mass spectrometry, coupled with a THERMO mass spectrometer detector ISQ Single Quadrupole Mass Spectrometer (THERMO Scientific Corp., Waltham, MA, USA)and were obtained by electron ionization (EI) at 70 eV, using a spectral range of m/z 50–1000.All of the microanalyses and spectral analyses were performed by the Micro Analytical Centers of Taif University-Saudi Arabia (IR spectra, HRMS), Cairo University (¹H-NMR, ¹³C-NMR), and National Research Center-Egypt (Mass spectra).

Khalifa M.E., Gobouri A.A., Kabli F.M., Altalhi T.A., Almalki A.S, & Mohamed M.A. (2018). Synthesis, Antibacterial, and Anti HepG2 Cell Line Human Hepatocyte Carcinoma Activity of Some New Potentially Benzimidazole-5-(Aryldiazenyl)Thiazole Derivatives. Molecules, 23(12), 3285.

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GC-MS Analysis of Samples

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Samples were analyzed in random order on a TRACE Ultra gas chromatograph (Thermo Scientific) equipped with an AS3000 II auto sampler and a DB-5MS Agilent column (30 m x 0.250 mm x 0.25 µm) with a 10 m Duraguard pre-column. The GC was connected to an ISQ Single Quadrupole mass spectrometer (Thermo Scientific). For each batch of 20 samples, a new deactivated glass liner (5 mm inner diameter, 8 mm outer diameter, 105 mm length; Thermo Scientific) with glass wool was used. Liners were shipped to CS-Chromatographie Service (Langerwehe, Germany) for cleaning and deactivation (GW INNO-Sil Plus). 1 μl of sample was injected at 300 °C and analyzed in split mode 10. The oven temperature was kept at 60 °C for 1 min, then increased by 15 °C min⁻¹ to 310 °C and kept at 310 °C for 10 min. The flow rate of the helium 5.0 carrier gas was set to 1 ml min⁻¹. Ionization and fragmentation was performed by an electron impact source at 70 eV and 280 °C. Five MS scans per second were recorded with a resolution of 866 at m/z 502.20 (FWHM = 0.53).

Hirth M., Liverani S., Mahlow S., Bouget F.Y., Pohnert G, & Sasso S. (2017). Metabolic profiling identifies trehalose as an abundant and diurnally fluctuating metabolite in the microalga Ostreococcus tauri. Metabolomics, 13(6), 68.

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Spectroscopic Analysis of Papaverine

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The 1D (¹H, ¹³C, APT and DEPT 135) and 2D (HMBC, COSY and HSQC) NMR spectra were acquired using standard pulse sequences on a Bruker model AMX 400 NMR spectrometer operating at 400 MHz and 100 MHz in ¹H NMR and ¹³C NMR, respectively. Results for the chemical shift (δ) were provided in parts per million units (ppm). J values for coupling constants were provided in Hertz (Hz). The HSQC, HMBC, APT and DEPT pulse sequences followed technical standards. For EI-MS, mass spectroscopy with a Thermo Scientific ISQ single-quadrupole mass spectrometer was used. For ESI-MS, a mass spectrometer from Thermo Scientific with a triple-quadrupole Access MAX system and Xcalibur 2.1 software was used. Sephadex LH-20 and silica gel (E. Merck, 70–230 mesh) column chromatography were used for obtaining pure metabolites. Analytical-grade chemicals were used in isolation and purification. Merck TLC sheets and silica gel G 254 F sheets were used for TLC (E. Merck, Germany). Solvent systems S1, ethyl acetate–methanol–ammonia (95:5:5), and S2, EtAc-MeOH-H₂O (4:1:5, v/v), were utilized for TLC analysis. Dragendorff’s and anisaldehyde/sulfuric acid spray reagents and a UV lamp were used to visualize the TLC plates (at 254 and 365 nm). Papaverine was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Eliwa D., Kabbash A., El-Aasr M., Tawfik H.O., Batiha G.E., Mahmoud M.H., De Waard M., Eldehna W.M, & Ibrahim A.R. (2023). Papaverinol-N-Oxide: A Microbial Biotransformation Product of Papaverine with Potential Antidiabetic and Antiobesity Activity Unveiled with In Silico Screening. Molecules, 28(4), 1583.

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