Db 5 capillary column

Manufactured by Agilent Technologies

Sourced in United States, Japan, Germany, Italy

The DB-5 capillary column is a widely used gas chromatography (GC) column designed for the separation and analysis of a wide range of organic compounds. It features a nonpolar stationary phase, making it suitable for the separation of a variety of analytes. The column is constructed with a fused silica capillary and a bonded phenyl-methyl siloxane stationary phase. This combination provides efficient separation and reliable performance for a diverse range of applications.

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DB-5 column (97 citations) DB-5 fused silica capillary column (18 citations) 30-m DB-5 capillary column (6 citations) DB-5 ms Agilent fused silica capillary column (4 citations) 30-m DB-35MS 5-m Duraguard capillary column (3 citations) DB‐5 quartz capillary column (2 citations)

162 protocols using db 5 capillary column

Alkaloid Extraction and Analysis by GC-FID and GC-MS

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The extraction of alkaloids was analyzed on a Younglin Acm 600 instrument with an FID detector operated with a split/splitless injector (Younglin, Korea) and DB-5 capillary column, 30 m × 0.25 mm i.d., 0.25 μm film thicknesses (Agilent, USA).
Carrier gas: He, linear velocity (u): 30 cm/sec, flow: 0.8 ml/min Injection temperature: 290 °C. Injection volume: 1.0 μl. Injection mode: Split (1:50). Temperature program: 50 °C for min, rising at 3 °C /min to 240 °C, then rising at 15 °C/min to 300 °C, held at 300 °C for 3 min. FID (290 °C): H₂ flow: 50 ml/min; air flow: 400 ml/min.
GC/MS analysis was performed on an Agilent 6890/5973 N instrument and DB-5 capillary column (30 m × 0.25 mm i.d., 0.25 μm film thickness). Carrier gas: He, Linear velocity (u): 32.4 cm/sec, flow: 0.8 ml/min. Injection temperature: 290 °C. Injection volume: 1.0 μl. Injection mode: split (1:10). Temperature program: 50 °C, for 5 min, rising at 3 °C/min to 240 °C, then rising at 15 °C/min to 300 °C, held at 300 °C for 3 min. MS interface temperature: 290 °C, MS mode: EI, Ionization voltage: 70 eV; mass range: 40-500 u; scan speed: 3.18 scans/sec; interval: 0.50 sec (2 Hz). Data handling was conducted using a Chem. Station (Agilent).

Kianbakht S, & Hashem Dabaghian F. (2016). Sophora alopecuroides L. var. alopecuroides alleviates morphine withdrawal syndrome in mice: involvement of alkaloid fraction and matrine. Iranian Journal of Basic Medical Sciences, 19(10), 1090-1095.

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Antennal Electrophysiological Response of Nicrophorus vespilloides to Methyl Geranate

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The electrophysiological response of N. vespilloides male antennae towards methyl geranate was tested using GC-EAD. The same GC-EAD set-up as in ref. 81 was used, but the GC was equipped with a DB-5 capillary column (30 m × 0.25 mm i.d., 25 μm film thickness, J & W Scientific, Folsom, CA, USA) and the oven temperature was raised from 50 °C to 220 °C at a rate of 10 °C per min. The final temperature was held for 2 min. The responses of three male antennae were tested to 50 ng of synthetic methyl geranate (100 ng μl⁻¹ pentane, 1 μl injected, Split FID:EAD=1:1, mixture of isomers, Santa Cruz Biothechnology, USA). The antennae were cut at their base and at the tip. The excised antenna was mounted between two glass electrodes filled with insect Ringer solution (see ref. 81 for further details).

Engel K.C., Stökl J., Schweizer R., Vogel H., Ayasse M., Ruther J, & Steiger S. (2016). A hormone-related female anti-aphrodisiac signals temporary infertility and causes sexual abstinence to synchronize parental care. Nature Communications, 7, 11035.

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Quantification of Target PCBs using GC-μECD

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Target PCBs were determined using gas chromatograph (Agilent 6890 Series, Agilent, Palo Alto, CA) equipped with micro electron capture detector. Separation was performed on a DB-5 capillary column (60 m length, 0.25 mm i.d., 0.25 μm film thickness, J&W Scientific, USA). The initial oven temperatures was set at 80 °C for 2 min then increased to 185 °C at a rate of 30 °C min⁻¹ (3 min), then at 1.5 °C min⁻¹ to 230 °C (held for 15 min) and finally to 270 °C at a rate of 5 °C min⁻¹ (held for 25 min). Nitrogen as carrier gas was used at 1.8 mL min⁻¹ flow rate. Injector was set at 270 °C and split-less mode. Detector was set at 300 °C. Confirmation of target PCB congeners were carried out using a Varian Gas Chromatograph CP-3800 (Varian, CA, USA) coupled to ion trap mass spectrometry detector (Varian Saturn 2000, Varian, CA, USA). The mass spectrums were acquired in MS/MS mode. 2 μL of each sample was injected in a programmable temperature vaporizing (PTV) injector (split-less mode). A VF-5MS capillary column (55 m length, 0.25 mm i.d. and 0.25 μm film thicknesses, Factor Four ®, Varian, Palo Alto, CA, USA) was employed. The GC oven was programmed as: initial temperature 100 °C for 2 min then to 200 °C (held for 3 ° min) at a rate of 30 °C min⁻¹ and then to 230 °C (held for 15 min) at a rate of 3 °C min⁻¹ and finally to 270 °C (held for 15 min) at a 5 °C min⁻¹ [18 (link), 19 (link)].

Ahmadkhaniha R., Nodehi R.N., Rastkari N, & Aghamirloo H.M. (2017). Polychlorinated biphenyls (PCBs) residues in commercial pasteurized cows’ milk in Tehran, Iran. Journal of Environmental Health Science and Engineering, 15, 15.

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GC-MS Metabolite Derivatization Protocol

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Derivatization of GC-MS samples was conducted following a previous report (Peng et al., 2015 ) with minor modifications. In brief, each sample was added with 80μL of methoxyamine (15 mg/mL, in pyridine). The mixture was then vortexed for 2 minutes and incubated for 90 minutes at 37 °C. 20 μL of n-hexane and 80 μL of bis(trimethylsilyl) trifluoroacetamide (BSTFA) (with 1% of trimethylchlorosilane) were then added. The solution was vortexed for 2 minutes, then reacted for 60 minutes at 70 °C, and finally incubated at room temperature for 30 minutes before GC-MS.
1 μL derivatized solution was then injected into the GC-MS system (Agilent 7890A-5975C, USA) using splitless mode. A non-polar DB-5 capillary column (J&W Scientific, 30 m × 250 μm I.D.) was used to perform the separation with a constant flow rate of 1.0 mL/min with carrier gas of high purity helium. The programmed GC temperatures were as follows: 15°C/min, 50°C-125°C; 5°C/min, 125°C-210°C; 10°C/min, 210°C-270°C; 20°C/min, 270°C-305°C with a final maintenance for 5 minutes at 305°C. Full scan mode (mass-to-charge ratio range of 50 to 600) with the acquisition rate set at 20 spectrum/second was used. The filament bias was set as −70 V and the electron impact (EI) ion source was held at 230°C.

Xu Y., Wang W., Zhou J., Chen M., Huang X., Zhu Y., Xie X., Li W., Zhang Y., Kan H, & Ying Z. (2019). Metabolomics Analysis of a Mouse Model for Chronic Exposure to Ambient PM2.5. Environmental pollution (Barking, Essex : 1987), 247, 953-963.

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GC-MS Analysis of Extracted Samples

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Samples weighting 60 mg were extracted as previously described (Chen et al., 2017 (link)). After extraction, 1 μL prepared sample solution was injected into the Agilent 7890A-5975C gas chromatography-mass spectrometry (GC-MS) system (Agilent Corporation, United States). GC-MS analysis was carried out on a non-polar DB-5 capillary column (30 m × 250 μm I.D., J&W Scientific, Folsom, CA). High purity of helium was used as the carrier gas at a constant flow rate of 1.0 mL/min. The temperatures of injection and ion source were set to 305 and 230°C, respectively. Electron impact ionization (-70 eV) at full scan mode (m/z 30-600) was used, with an acquisition rate of 20 spectrum/second in the MS setting. QC sample was prepared by mixing aliquots of tissues samples to be a pooled sample.
Acquired data were analyzed by ChromaTOF software (v 4.34, LECO, St Joseph, MI). Internal standards and any known pseudo positive peaks were removed from the obtained data set. Data set was normalized using sum intensity of peaks in each sample. Obtained three-dimensional data sets included sample information, retention time, and peak intensities.

Chen Q., Jin Y., Zhang Z., Cao M., Wei G., Guo X., Zhang J., Lu X, & Tang Z. (2021). Ionomic and Metabolomic Analyses Reveal Different Response Mechanisms to Saline–Alkali Stress Between Suaeda salsa Community and Puccinellia tenuiflora Community. Frontiers in Plant Science, 12, 774284.

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Volatile Compounds Analysis by GC-MS

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The analysis of volatile compounds was conducted using the Agilent 6890 GC System equipped with the Agilent 5975 MS (Agilent Technologies, Santa Clara, CA, USA) and fitted with the DB-5 capillary column (30 m × 0.25 mm ID, 0.25 μm film thickness; J&W Scientific, Folsom, CA, USA). Ultrahigh purity helium (≥99.999%) was employed as the carrier gas with a constant flow rate of 1 mL·min⁻¹, and 1 μl extraction of sample was injected at 250°C in splitless mode. The oven temperature was programmed at 40°C for 2 min and then ramped to 220°C at a rate of 6 °C·min⁻¹ and held at 220°C for 5 min. The mass spectrometer was operated in the electron impact ionization mode at a voltage of 70 eV and ion source temperature at 230°C. Mass spectra were taken over an m/z range of 30–400. Retention indices were calculated after analyzing C₈–C₂₀n-alkane series under the same chromatographic conditions.

Zhang L., Mi S., Liu R.B., Sang Y.X, & Wang X.H. (2020). Evaluation of Volatile Compounds during the Fermentation Process of Yogurts by Streptococcus thermophilus Based on Odor Activity Value and Heat Map Analysis. International Journal of Analytical Chemistry, 2020, 3242854.

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GC-MS Analytical Protocol for Metabolite Characterization

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Each 2 μL aliquot of the derivatized solution was injected into the Agilent 7890A-5975C Gas Chromatograph-Mass Spectrometer (Agilent, California, USA) in splitless mode. Separation was carried out on a nonpolar DB-5 capillary column (30 m × 250 μm ID; J&W Scientific, Folsom, CA, USA), with high purity helium as the carrier gas at a constant flow rate of 1.0 mL/min. The GC temperature program began at 80°C, and the temperature was increased by increments of 10°C/min in an oven to 180°C. This was followed by incremental increases of 5°C/min to 240°C and 25°C/min to 290°C and then a final 11-minute maintenance at 290°C. The electron impact ion source was maintained at 260°C with a filament bias of −70 V. Full scan mode (m/z 50–600) was used with an acquisition rate of 20 spectra/s in the MS setting. A QC sample was prepared by mixing aliquots of all of the samples into a pooled sample, and this was then analyzed using the same method as the analytical samples. The QCs were injected at regular intervals (every ten samples) throughout the analytical process to provide a set of data from which repeatability could be assessed.

Zhao K., Zhang J., Xu Z., Xu Y., Xu A., Chen W., Miao C., Liu S., Wang Z, & Jia R. (2018). Metabolomic Profiling of Human Spermatozoa in Idiopathic Asthenozoospermia Patients Using Gas Chromatography-Mass Spectrometry. BioMed Research International, 2018, 8327506.

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Gas Chromatography-Mass Spectrometry Analysis

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The volatile composition analysis of the oil was carried out using a gas chromatography–mass spectrometry (GC-MS, 6890N GC and 5975 MS, Agilent Technologies, Santa Clara, CA, United States) system equipped with a split/splitless injector and a DB-5 capillary column (60-m length, 0.25-mm diameter, and 1-μm film thickness; J&W Scientific, Folsom, CA, United States). The injector and detector temperature was 260°C. The injection volume was 1 μl with a split ratio of 40:1. The oven conditions were 40°C (0.5 min) and then 4°C⋅min^–1 to 225°C (held for 13.25 min) for a total run time of 60 min. Helium was used as carrier gas at flow rate of 1.5 ml⋅min^–1. Inlet, ionizing source, and transfer line were kept at 250, 230, and 280°C, respectively. The mass spectrometry data were recorded in the scan mode from 40 to 400 m/z at 2 scans⋅s^–1 with an ionization energy of 70 eV.

Sun X., Yang H., Zhao W., Bourcier E., Baldwin E.A., Plotto A., Irey M, & Bai J. (2021). Huanglongbing and Foliar Spray Programs Affect the Chemical Profile of “Valencia” Orange Peel Oil. Frontiers in Plant Science, 12, 611449.

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GC-MS Analysis of Volatile Compounds

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The volatile components were injected at 240°C into the injection port of GC/MS (GCMS-QP2010, Shimadzu Co., Ltd., Kyoto, Japan) for 5 min. The volatile components were sampled for 1 min using a splitless mode, and then separated using a DB-5 capillary column (30 m×0.25 mm i.d.×0.25 μm, J&W Scientific, Inc., Folsom, CA, USA). Helium was used as the mobile phase gas. Analysis conditions were set to 0.8 mL/min of flow rate, 32.2 cm/min of linear velocity, and 31.9 KPa of column pressure. Oven temperature was increased to 220°C at 2.5°C/min after keeping for 5 min at 35°C, and kept for 31 min. For the MS analysis conditions, the temperatures of capillary direct interface and ion source were 220°C. The electron multiplier voltage and MS ionization voltage were 1,000 V and 70 eV. The mass range and scan rate were 40~ 350 m/z and 1/s, respectively. The volatile flavor component was determined by the consistency of the retention indices (RI) between the volatile components and reference material (15 (link)), or the comparison of mass spectra between volatile components and Wiley 8 (399,383 spectrum) mass spectral database (Shimadzu Co., Ltd.). RI was carried out under the same conditions as the samples using C₆~C₂₄ (n-alkane, Aldrich, Milwaukee, MN, USA).

Kim J.H., Cha J.Y., Shin T.S, & Chun S.S. (2018). Volatile Flavor Components of Blended Tea with Fermented Tea and Herbs. Preventive Nutrition and Food Science, 23(3), 245-253.

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GC-MS Analysis of Honeysuckle Seed Oil

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The fatty acid composition in blue honeysuckle seed oil was analyzed using GC–MS. To increase the volatility of the seed oil, pre-column derivatization was used. In a 10 mL screw-cap glass tube, 30 µL of seed oil was dissolved in a mixed solution of n-hexane and benzene (1:1, v/v) and gently shaken. Then, 2 mL of 0.5 mol/L KOH in methanol was added, and the mixture was allowed to stand for 30 min. Subsequently, 5 mL of distilled water was added to separate the organic phase solution (n-hexane), causing it to rise to the upper layer of the tube. The top layer solution was diluted 20 times and analyzed by GC–MS on an Agilent 6890 GC system equipped with a 5973 N mass selective detector (Agilent Technologies Inc., Wilmington, DE, USA) using a DB-5 capillary column (60 m × 0.25 mm id, 0.25 µm film thickness, J&W Scientific, CA, USA). The GC injection port temperature was 250 °C, and helium was used as the carrier gas at a flow rate of 1.0 mL/min. The injection volume was 1.0 µL, and the injection split ratio was 1:10. The oven temperature was started at 180 °C and held for 5 min, and then ramped to 240 °C at 3 °C/min and held for 8 min. The total program time was 32 min. The ion source temperature was 230 °C. The MS detection was operated at 70 eV with a scan range of 50–550 m/z.

Sun J., Li D., Huyan W., Hong X., He S., Huo J., Jiang L, & Zhang Y. (2024). Blue honeysuckle seeds and seed oil: Composition, physicochemical properties, fatty acid profile, volatile components, and antioxidant capacity. Food Chemistry: X, 21, 101176.

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