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Duraguard

Manufactured by Agilent Technologies
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The DuraGuard is a protective cover designed for Agilent's analytical laboratory equipment. It is engineered to provide a durable and reliable safeguard for the instruments, helping to maintain their performance and extend their lifespan.

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Lab products found in correlation

Mtbstfa, by Merck Group (3 mentions) Agilent 7890b 5977b, by Agilent Technologies (3 mentions) Urea standard, by Merck Group (2 mentions) Pyridine, by Merck Group (2 mentions) Pegasus 4d tofms, by Leco (1 mentions) 7890b gc ms system, by Agilent Technologies (1 mentions) Masshunter quantitative analysis software, by Agilent Technologies (1 mentions)

5 protocols using duraguard

1

Comprehensive GC/MS Metabolite Profiling

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GC/MS was performed using a Pegasus 4D TOFMS (LecoCorp) equipped with an Agilent 7890 GC and a CTC CombiPAL autosampler. A DB-1 capillary column (30 m × 250 μm (i.d.) × 0.25 μm) with DuraGuard (Agilent Technologies J&W) was used. Helium flow rate was set at 1.5 mL/min and the injection volume was 1 μL with injector split ratio 1:20. For GC, the temperature was 220°C for front inlet and 280°C for transfer line. Column temperature was programed to be at 70°C for 0.2 min, ramped at 15°C/min to 270°C and then at 40°C/min to 310°C, and finally hold at 310°C for 8 min. For MS, the detector voltage was 1,600 V with an acquisition delay of 200 seconds. To facilitate metabolite identification, alkane standard mix (C10-C40; Sigma) and FAME (fatty acid methyl esters) standards (C8-C28; Sigma) were analyzed using the same settings, so that the GC retention time could be converted to two types of retention index (RI), Kovats index and Fiehn index, respectively.
Li C., Li P., Tan Y.M., Lam S.H., Chan E.C, & Gong Z. (2016). Metabolomic Characterizations of Liver Injury Caused by Acute Arsenic Toxicity in Zebrafish. PLoS ONE, 11(3), e0151225.
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2

Sucrose Extraction and Derivatization for GC-MS

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Sucrose extraction from the freeze-dried sample and its derivatization for GC/MS analysis were essentially carried out as previously described71 (link). GC-EIMS used in this study was a 7890B GC-MS system (Agilent Technologies) equipped with a DB-5 MS + Dura Guard (30 m × 0.25 mm i.d., film thickness of 0.5 μm and 10 m Dura Guard, Agilent Technologies). The injection temperature was 250 °C, and the helium gas flow rate through the column was 0.9 mL/min. The column temperature was held at 60 °C for 1 min and then was raised by 10 °C/min to 325 °C and was held there for 10 min isothermally. The retention time for sucrose-8TMS was 16.4 min under the condition.
Hiruma K., Aoki S., Takino J., Higa T., Utami Y.D., Shiina A., Okamoto M., Nakamura M., Kawamura N., Ohmori Y., Sugita R., Tanoi K., Sato T., Oikawa H., Minami A., Iwasaki W, & Saijo Y. (2023). A fungal sesquiterpene biosynthesis gene cluster critical for mutualist-pathogen transition in Colletotrichum tofieldiae. Nature Communications, 14, 5288.
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3

GC-MS Analysis of Metabolite Extraction

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Following treatment, metabolites were extracted by scraping cells in 80% methanol and vortexing. Extracts were spin-clarified, and supernatants were collected and dried by vacuum centrifugation at 4°C (Labconco). Dried supernatant pellets were resuspended in 15 ul of 10 mg/ml methoxylamine in pyridine (Sigma), incubated for 30 min at 37°C, and derivatized by silylation with 35 μl of N-Methyl-N-tert-butyldimethylsilyltrifluoroacetamide (MTBSTFA, Sigma) for 1h at 70°C. Derivatized samples were spin clarified and 1 μl per sample was injected in splitless mode into an Agilent 5977B/7890B GC-MS with a DB-5ms capillary column with DuraGuard (Agilent Technologies), using chromatographic methods as previously described61 (link). Succinate was confirmed using 13C4-succinate (m+ 4) standard, and had a retention time of 14.1 min, quantifying ion of 289 + n, and qualifying ion 331 + n. Data were analyzed in batch format using Mass Hunter Quantitative Analysis software (Agilent Technologies). Data are shown as total ion counts normalized relative to total ion abundance and corrected for natural abundance and normalized to cell number upon plating.
Mills E.L., Harmon C., Jedrychowski M.P., Xiao H., Garrity R., Tran N.V., Bradshaw G.A., Fu A., Szpyt J., Reddy A., Prendeville H., Danial N.N., Gygi S.P., Lynch L, & Chouchani E.T. (2021). UCP1 governs liver extracellular succinate and inflammatory pathogenesis. Nature metabolism, 3(5), 604-617.
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4

Polar Metabolites Analysis in Islets

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Polar metabolites were extracted in 100 µl of 80% methanol (GC-grade, Thermo Optima) added to 100 µl of spent media from human or mouse islets (60 islets per 2ml) and lysed by sonication. Extracts were spin-clarified, and supernatants were collected and dried by vacuum centrifugation at 4°C (Labconco). Dried supernatant pellets were resuspended in 10 mg per ml of methoxylamine in pyridine (Sigma), incubated for 30 min at 37°C, and derivatized by silylation with 70 µl of N-Methyl-N-tert-butyldimethylsilyltrifluoroacetamide (MTBSTFA, Sigma) for 1h at 70°C. After spin clarifying, 1 µl of derivatized sample was injected in splitless mode into an Agilent 5977B/7890B GC-MS with a DB-5ms capillary column with DuraGuard (Agilent Technologies), using chromatographic methods as previously described40 (link). Data are shown either as concentrations (ng/ml) using a urea standard (Sigma) calibration curve or as values relative to PBS-treated controls. Between sample differences were corrected for by a factor of median chromatographic peak area.
Fu A., Alvarez-Perez J.C., Avizonis D., Kin T., Ficarro S.B., Choi D.W., Karakose E., Badur M.G., Evans L., Rosselot C., Bridon G., Bird G.H., Seo H.S., Dhe-Paganon S., Kamphorst J.J., Stewart A.F., Shapiro A.M., Marto J.A., Walensky L.D., Jones R.G., Garcia-Ocana A, & Danial N.N. (2020). Glucose-dependent partitioning of arginine to the urea cycle protects β-cells from inflammation. Nature metabolism, 2(5), 432-446.
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5

Polar Metabolites Analysis in Islets

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Polar metabolites were extracted in 100 µl of 80% methanol (GC-grade, Thermo Optima) added to 100 µl of spent media from human or mouse islets (60 islets per 2ml) and lysed by sonication. Extracts were spin-clarified, and supernatants were collected and dried by vacuum centrifugation at 4°C (Labconco). Dried supernatant pellets were resuspended in 10 mg per ml of methoxylamine in pyridine (Sigma), incubated for 30 min at 37°C, and derivatized by silylation with 70 µl of N-Methyl-N-tert-butyldimethylsilyltrifluoroacetamide (MTBSTFA, Sigma) for 1h at 70°C. After spin clarifying, 1 µl of derivatized sample was injected in splitless mode into an Agilent 5977B/7890B GC-MS with a DB-5ms capillary column with DuraGuard (Agilent Technologies), using chromatographic methods as previously described40 (link). Data are shown either as concentrations (ng/ml) using a urea standard (Sigma) calibration curve or as values relative to PBS-treated controls. Between sample differences were corrected for by a factor of median chromatographic peak area.
Fu A., Alvarez-Perez J.C., Avizonis D., Kin T., Ficarro S.B., Choi D.W., Karakose E., Badur M.G., Evans L., Rosselot C., Bridon G., Bird G.H., Seo H.S., Dhe-Paganon S., Kamphorst J.J., Stewart A.F., Shapiro A.M., Marto J.A., Walensky L.D., Jones R.G., Garcia-Ocana A, & Danial N.N. (2020). Glucose-dependent partitioning of arginine to the urea cycle protects β-cells from inflammation. Nature metabolism, 2(5), 432-446.
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