5977a mass spectrometer

Manufactured by Agilent Technologies

Sourced in United States, United Kingdom

The 5977A mass spectrometer is a laboratory instrument designed for high-performance mass analysis. It combines a robust electron ionization (EI) ion source with a high-efficiency quadrupole mass analyzer to provide accurate and reliable mass measurements. The 5977A is capable of detecting and identifying a wide range of chemical compounds with high sensitivity and selectivity.

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Lab products found in correlation

45 protocols using 5977a mass spectrometer

GC-FID and GC-MS Analysis of Terpenes

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Analyses were performed on an HP 6890 GC system (Agilent Technologies, Santa Clara, CA, USA) with flame ionization detection. An HP-5 capillary column (30 m × 0.25 mm × 0.25 µm) was used to separate the terpenes. The injection and detector temperatures were maintained at 250 °C for the BP extract. The injection volume was 1 µL with a split ratio (1:10). Helium, hydrogen, and air were used as carrier gases. The oven temperature was maintained at 50 °C for 2 min and then increased at a heating rate of 4 °C for 5 min until a final temperature 250 °C of was reached.
GC–MS analysis was performed on a 7890B GC system (Agilent Technologies, Santa Clara, CA, USA) with a 5977A mass spectrometer (Agilent Technologies, Santa Clara, CA, USA). An HP-5 capillary column (30 m × 0.25 mm × 0.25 µm) was used to separate the terpenes. The injection and detector temperature were 250 °C for BP extracts. The injection volume was 1 µL with a split ratio (1:10). Helium, hydrogen, and air were used as carrier gases. The oven temperature was maintained at 60 °C for 2 min and then increased at a heating rate of 20 °C/min until it reached a final temperature of 280 °C.

Lee H.Y, & Ko M.J. (2021). Thermal decomposition and oxidation of β-caryophyllene in black pepper during subcritical water extraction. Food Science and Biotechnology, 30(12), 1527-1533.

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GC-MS Analysis of Amino Acids and Fatty Acids

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GC/MS analysis was performed on an Agilent 7890B GC system equipped with a DB-5MS capillary column (30 m, 0.25 mm i.d., 0.25 µm-phase thickness; Agilent J&W Scientific), connected to an Agilent 5977A Mass Spectrometer operating under ionization by electron impact (EI) at 70 eV. Helium flow was maintained at 1 mL/min. The source temperature was maintained at 230°C, the MS quad temperature at 150°C, the interface temperature at 280°C, and the inlet temperature at 250°C. For GC/MS analysis of amino acids, 1 µL was injected at 1:40 split ratio. The column was started at 80°C for 2 min, increased to 280°C at 7°C/min, and held for 20 min. For GC/MS analysis of fatty acid methyl esters (FAME) and sugar derivatives, 1 µL was injected splitless. The column was started at 80°C for 2 min, increased to 280°C at 10°C/min, and held for 12 min.

Long C.P, & Antoniewicz M.R. (2014). Quantifying Biomass Composition by Gas Chromatography/Mass Spectrometry. Analytical chemistry, 86(19), 9423-9427.

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Cannabis Compound Identification by GC-MS

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GC-MS analyses were carried out using an Agilent 7890B gas chromatograph coupled to a 5977A mass spectrometer (electron multiplier potential 2 kV, filament current 0.35 mA, electron energy 70 eV, and the spectra were recorded over the range of m/z 40 to 500). An Agilent 7683 autosampler was used for sample introduction. A 1 μL aliquot of each sample was injected into the GC–MS using a 1:10 split-ratio injection mode. Helium was used as the carrier gas at a constant flow of 1.1 mL s⁻¹. An isothermal hold at 50 °C was maintained for 2 min, followed by a heating gradient of 6 °C min⁻¹ to 300 °C, and the final temperature was held for 4 min. A 3 min solvent delay was applied. A 30 m, 0.25 mm ID, 5% cross-linked phenylmethyl siloxane capillary column (HP-5MS) with 0.25 μm film thickness was used for separation, and the injection port temperature was 220 °C. The MS interface temperature was 280 °C.
Peak assignments were performed with a spectral library (NIST 14.0) and compared with MS data obtained from the injection of standards, purchased from LGC Standards (Teddington, UK). For identification and partial quantification, 10 µg of the aforementioned phytocannabinoid and terpenoid standards were injected to the GC–MS.

Namdar D., Voet H., Ajjampura V., Nadarajan S., Mayzlish-Gati E., Mazuz M., Shalev N, & Koltai H. (2019). Terpenoids and Phytocannabinoids Co-Produced in Cannabis Sativa Strains Show Specific Interaction for Cell Cytotoxic Activity. Molecules, 24(17), 3031.

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Derivatization of Dried Residues for GC-MS Analysis

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the dried residues were derivatized with methoxylamine hydrochloride and propionic anhydride sequentially adapted from a previously described method (12 (link)). 25 μL of methoxylamine hydrochloride (2% (w/v) in pyridine) was added to the dried residues and incubated for 60 minutes at 70°C. After centrifugation for 2 minutes at 12,000 g, 50 μL propionic anhydride was added and incubated for 30 minutes at 60°C prior to another round of drying under nitrogen. The dried residues were resuspended with 55ul pure ethyl acetate and transferred to GC vials for analysis. GC/MS analysis was conducted using the Agilent 7890B GC system and Agilent 5977A Mass Spectrometer. Specifically, 1 μL of the derivatized sample was injected into the GC column. GC temperature gradient started at 90°C, increased to 260°C at the speed of 9°C per minute and further increased to 290 °C at the speed of 30°C per minute. Then temperature was held at 290°C for 5 minutes with a run time of 24.9 minutes. Ionization was conducted by EI at 70 eV with Helium flow at 1 mL/min. Temperatures of the source, the MS quad, the interface, and the inlet were maintained at 230°C, 150°C, 280°C, and 250°C, respectively. Mass spectra were recorded in mass scan mode from m/z 50 to 700.

Tong W., Hannou S.A., Wang Y., Astapova I., Sargsyan A., Monn R., Thiriveedi V., Li D., McCann J.R., Rawls J.F., Roper J., Zhang G.F, & Herman M.A. (2022). “The Intestine is a Major Contributor to Circulating Succinate in Mice”. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 36(10), e22546.

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GC-MS Analysis of Amino Acids and Glucose Labeling

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GC-MS analysis was performed on an Agilent 7890B GC system equipped with a DB-5MS capillary column (30 m, 0.25 mm i.d., 0.25 μm-phase thickness; Agilent J&W Scientific), connected to an Agilent 5977A Mass Spectrometer operating under ionization by electron impact (EI) at 70 eV. Helium flow was maintained at 1 mL/min. The source temperature was maintained at 230°C, the MS quad temperature at 150°C, the interface temperature at 280°C, and the inlet temperature at 250°C. GC-MS analysis of tert-butyldimethylsilyl (TBDMS) derivatized proteinogenic amino acids was performed as described in (Long and Antoniewicz, 2014b (link)). Labeling of glucose was determined using the aldonitrile propionate derivatization method described in (Antoniewicz et al., 2011 (link)).

Crown S.B., Long C.P, & Antoniewicz M.R. (2016). Optimal tracers for parallel labeling experiments and 13C metabolic flux analysis: A new precision and synergy scoring system. Metabolic engineering, 38, 10-18.

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Isotopic Labeling Analysis by GC-MS

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GC-MS analysis was performed on an Agilent 7890B GC system equipped with a DB-5MS capillary column (30 m, 0.25 mm inner diameter, 0.25 μm-phase thickness; Agilent J&W Scientific), connected to an Agilent 5977A Mass Spectrometer operating under ionization by electron impact (EI) at 70 eV. Helium flow was maintained at 1 ml min⁻¹. The source temperature was maintained at 230 °C, the MS quad temperature at 150 °C, the interface temperature at 280 °C and the inlet temperature at 250 °C. GC-MS analysis of tert-butyldimethylsilyl (TBDMS) derivatized proteinogenic acids was performed to measure isotopic labelling30 (link). Mass isotopomer distributions were obtained by integration31 (link) and corrected for natural isotope abundances32 (link).

Long C.P., Au J., Sandoval N.R., Gebreselassie N.A, & Antoniewicz M.R. (2017). Enzyme I facilitates reverse flux from pyruvate to phosphoenolpyruvate in Escherichia coli. Nature Communications, 8, 14316.

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GC-MS Analysis of Volatile Organic Compounds

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The analytical system was composed of an Agilent 7890 B gas chromatograph coupled with an Agilent 5977 A mass spectrometer with a MPS, a thermal desorption unit (TDU) and a cooled injection system (CIS) (Gerstel). The data acquisition software MSD Chemstation F.01.00 (Agilent Technologies, Les Ulis, France) was used to program the GC-MS. The gas chromatograph was fitted with a VF-WAXms fused silica capillary column (30 m × 0.25 mm × 0.25 µm, Agilent) and was used with helium as carrier gas at 0.8 mL·min⁻¹.
The tube was desorbed in the TDU at 220 °C (suitable temperature for both bars, recommended by the supplier) for 5 min. After desorption, VOCs were focalized on the CIS at −10 °C during 2 min, ramped to 250 °C at a heating rate of 12 °C per second, and finally held for 2.5 min in splitless mode to ensure complete desorption of analytes. The column temperature was initially kept at 40 °C for 7 min and then increased from 40 °C to 240 °C at 8 °C·min⁻¹, the temperature was maintained during 3 min. Then, the sample was introduced into the ion source of the Agilent 5977 A mass spectrometer. The transfer line temperature was set at 250 °C and ion source temperature at 230 °C. Ions were generated by a 70 eV electron beam. Masses were acquired from m/z 33–500 amu.

Berrou K., Dunyach-Remy C., Lavigne J.P., Roig B, & Cadiere A. (2019). Comparison of Stir Bar Sorptive Extraction and Solid Phase Microextraction of Volatile and Semi-Volatile Metabolite Profile of Staphylococcus aureus. Molecules, 25(1), 55.

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GC-MS Analysis of Volatile Metabolites

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The analysis of volatile compounds was carried out on an Agilent 7890A gas chromatograph (GC) equipped with a split–splitless injector. The GC was coupled to an Agilent 5977A mass spectrometer (MS) with a quadrupole type mass filter and an electronic impact ionization mode. The GC was equipped with an upstream Gerstel MPS2-XL sampler and with a liquid injection module allowing the analysis of samples. The parameters of the sample changer were as follows: an injection volume of 1 μL carried out in splitless mode at 250 °C. The volatile metabolites were separated on a DB-5 MS capillary column (30 m × 0.25 mm × 0.25 μm; Agilent J&W). The initial temperature of the oven was 140 °C, maintained for 5 min. The temperature was increased by steps of 4 °C/min until reaching 240 °C, then was maintained for 20 min; the transfer line was at 280 °C. The MS was used in scan mode with a solvent delay of 4.6 min; the mass range used to identify the metabolites was between 29 and 450 m/z.

Blais A., Lan A., Boluktas A., Grauso-Culetto M., Chaumontet C., Blachier F, & Davila A.M. (2022). Lactoferrin Supplementation during Gestation and Lactation Is Efficient for Boosting Rat Pup Development. Nutrients, 14(14), 2814.

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Metabolite Derivatization and GC-MS Analysis

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Dried residues were derivatized with methoxylamine hydrochloride and tertbutyldimetheylchloros (TBDMS) sequentially as previously described [3]. Specifically, 25 μL of methoxylamine hydrochloride (2% (w/v) in pyridine) was added to the dried residues and incubated for 90 minutes at 40°C before the addition of 35 μL of TBDMS and incubation for 30 minutes at 60°C. The samples were then centrifuged for 2 minutes at 12,000 g and the supernatants of derivatized samples were transferred to GC vials for further analysis. GC/MS analysis was conducted as previously described using an Agilent 7890B GC system and Agilent 5977A Mass Spectrometer [3]. Specifically, 1 μL of the derivatized sample was injected into the GC column. GC temperature gradient started at 80°C for 2 minutes, increased to 280°C at the speed of 7°C per minute, and held at 280°C for until the completion of a run time of 40 minutes. Ionization was conducted by electron impact (EI) at 70 eV with Helium flow at 1 mL/min. Temperatures of the source, the MS quadropole, the interface, and the inlet were maintained at 230°C, 150°C, 280°C, and 250°C respectively. Mass spectra were recorded in mass scan mode from m/z 50 to 700.

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Derivatization and GC-MS Analysis of Metabolites

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Dried sample residues were derivatized with methoxylamine hydrochloride and trimethylsilyl (TMS) sequentially. Specifically, 25 μL of methoxylamine hydrochloride (2% (w/v) in pyridine) was added to the dried residues and incubated for 90 minutes at 40°C before the addition of 35 μL of TMS and incubation for 30 minutes at 60°C. The samples were then centrifuged for 2 minutes at 12,000 g and the supernatants of derivatized samples were transferred to GC vials for further analysis. GC/MS analysis was conducted using Agilent 7890B GC system and Agilent 5977A Mass Spectrometer. To be specific, 1 ul of the derivatized sample was injected into the GC column. GC temperature gradient start at 80°C for 2 minutes, increased to 280°C at the speed of 7°C per minute, and held at 280°C for until the completion of a run time of 40 minutes. Ionization was conducted EI at 70 eV with Helium flow at 1 mL/min. Temperatures of the source, the MS quadropole, the interface, and the inlet were maintained at 230°C, 150°C, 280°C, and 250°C respectively. Mass spectra were recorded in mass scan mode from m/z 50 to 700.

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