Extracellular 13C-labeling dynamics were analyzed by gas chromatography mass spectrometry (GC-MS). The GC-MS measurements were carried out on a GC (HP 6890, Hewlett Packard, Paolo Alto, CA, USA) equipped with an HP5MS capillary column (5% phenyl-methyl-siloxane diphenylpolysiloxane, 30 m × 0.25 mm × 0.25 μm, Agilent Technologies, Waldbronn, Germany), electron impact ionization at 70 eV, and a quadrupole detector (Agilent Technologies). The injection volume was 1 μl (7683B Autosampler, Agilent, Waldbronn, Germany; PTV-Injektor, Gerstel, Mühlheim a. d. Ruhr, Germany). Helium was used as carrier gas at a flow rate of 1.1 ml/min for analysis of lactate and amino acids or 0.7 ml/min for pyruvate analysis. The following temperature gradient was applied for lactate and amino acid analysis: 135°C for 7 min, 10°C/min up to 162°C, 7°C/min up to 170°C, 10°C/min up to 325°C, 325°C for 2.5 min; inlet temperature: 140°C and heating with 720°C/min up to 320°C; interface temperature 320°C; quadrupole temperature 150°C. The temperature gradient for pyruvate analysis was as follows: 70°C for 1 min, 1°C/min up to 75°C, 5°C/min up to 315°C, 25°C/min up to 340°C, 340°C for 5 min; inlet temperature: 70°C and heating with 360°C/min up to 360°C; interface temperature 320°C; quadrupole temperature 280°C.
7683b autosampler
Manufactured by Agilent Technologies
Sourced in United States, Germany
The 7683B autosampler is a laboratory instrument designed to automatically inject liquid samples into an analytical instrument, such as a gas chromatograph or liquid chromatograph, for analysis. The 7683B autosampler can hold and process multiple samples in a sequential manner, enabling efficient and consistent sample introduction without the need for manual intervention.
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Lab products found in correlation
Hp 5ms capillary column, by Agilent Technologies (1 mentions)
Hp 6890, by Hewlett-Packard (1 mentions)
7890 gas chromatograph, by Agilent Technologies (1 mentions)
Gc ms system, by Agilent Technologies (1 mentions)
Zb 1ms, by Phenomenex (1 mentions)
Chemstation software, by Agilent Technologies (1 mentions)
Gc 6890, by Agilent Technologies (1 mentions)
5975c msd, by Agilent Technologies (1 mentions)
Fused silica capillary column, by Agilent Technologies (1 mentions)
Hp 5ms column, by Agilent Technologies (1 mentions)
Db 5ms column, by Agilent Technologies (1 mentions)
Gc ms, by Agilent Technologies (1 mentions)
Micromass gct premier, by Waters Corporation (1 mentions)
Msd chemstation, by Agilent Technologies (1 mentions)
7890a network gc system, by Agilent Technologies (1 mentions)
5975c network mass selective detector, by Agilent Technologies (1 mentions)
Methanol, by Merck Group (1 mentions)
1 propanol, by Merck Group (1 mentions)
Isobutyl alcohol, by Merck Group (1 mentions)
Isoamyl alcohol, by Merck Group (1 mentions)
12 protocols using 7683b autosampler
1
GC-MS Analysis of Extracellular 13C-Labeling
2
GC/MS Analysis of Fatty Acid Esters
3
GC-MS Analysis of Purified Fractions
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The partially purified active fractions were analyzed by GC–MS. The analyses of the compounds in the active fractions were run on a GC–MS system (Agilent GC: 6890, with a 7683B Autosampler). The fused-silica Rxi-5Sil MS capillary column (30 m 0.25 mm ID, film thickness of 0.25 mm) was directly coupled to an Agilent variant. Oven temperature was programmed (35 °C for 5 min, then 35–300 °C at 10 °C/min) and subsequently, held isothermal for 20 min. The injector port; was 250 °C, the transfer line: 290 °C, splitless. Volume injected: 0.2 ml and the column flow rate was 1 ml/min of 1 mg/ml solution (diluted in chloroform). The peaks of components in gas chromatography were subjected to mass-spectral analysis.
The MS was a LECO Pegasus 4D recording with a EI-source at − 70 eV; the solvent delay was 9 min. Scan time 1.5 s; acquisition rate 10 spectra/second; mass range 50–1000 amu; detector voltage 1800 V, and Ion source temperature: 250 °C. Data were recorded in TIC mode. The software adopted to handle the mass spectra and chromatograms was an Agilent chemstation software. The constituents were identified after comparing with available data in the GC–MS library in the literatures.
The MS was a LECO Pegasus 4D recording with a EI-source at − 70 eV; the solvent delay was 9 min. Scan time 1.5 s; acquisition rate 10 spectra/second; mass range 50–1000 amu; detector voltage 1800 V, and Ion source temperature: 250 °C. Data were recorded in TIC mode. The software adopted to handle the mass spectra and chromatograms was an Agilent chemstation software. The constituents were identified after comparing with available data in the GC–MS library in the literatures.
4
Quantification of PAH Degradation in Compost
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Residual PAH-concentration from compost samples was recovered by extraction using automated Soxhlet technique based on EPA method 3541 [26 ] and dichloromethane (≥99.5%, Sigma-Aldrich) as solvent [20 (link)]. Briefly, 10 g of compost samples was transferred into a cellulose thimble and subjected to Soxhlet extraction. For each sample, a triplicate was prepared and extracted. The PAHs present in the extracts was quantified on GC/MS Agilent 7860GC system and 5975C MSD, equipped with a 7683B autosampler (Agilent Technologies Inc., California, USA). The column used was Agilent HP-5 MS ultra-inert 30 m × 0.25 mm x 0.25 μm film thickness (Agilent Technologies Inc., California, USA) and the GC-MSD conditions used for quantification were based on the optimised method described by Agilent Application Note (https://www.agilent.com/cs/library/applications/application-optimized-gc-ms-analysis-for-PAHs-in-challenging-matrices-8890-5977b-single-quadrupole-gc-ms-5994-0499en-agilent.pdf ). Prior samples analysis, the GC-MSD was calibrated with 100 μg/mL PAH standards (Sigma Aldrich Ltd).
The percent PAH degradation was calculated as follows:
The percent PAH degradation was calculated as follows:
5
GC-MS Analysis of Biological Extracts
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GC-MS analysis of the biological extracts was performed on a Agilent 7890A GC split/split less injector (280°C) linked to a Agilent 5975C MSD (electron voltage 70eV, source temperature 230°C, quad temperature 150°C multiplier voltage 1200V, interface temperature 310°C). The acquisition was controlled by a HP Compaq computer using Chemstation software, initially in full scan mode (50–600 amu/sec) or in selected ion mode (30ions 0.7cps 35ms dwell) for greater sensitivity. The sample (1ul) in diethyl ether was injected by an Agilent7683B auto sampler and the split opened after 1 minute. Separation was performed on an Agilent fused silica capillary column (30m x 0.25mm i.d) coated with 0.25um dimethyl polysiloxane (HP-5) phase. The GC was temperature programmed from 50–310°C at 5°C min and held at final temperature for 10 minutes with Helium as the carrier gas (flow rate of 1ml/min, initial pressure of 50kPa, split at 30 mls/min). Peaks were identified and labelled after comparison of their mass spectra with those of the NIST05 library if > 90% fit or from their elution order from biochemical literature.
6
GC-ECD Analysis of Organic Compounds
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The residue analysis was performed using the Agilent 6890N GC (Wilmington, NC, USA) equipped with an Agilent 7683B autosampler (Shanghai, China) and an electron capture detector (ECD) (Wilmington, NC, USA). Separation of the analytes was conducted using an HP-5 ((5%-Phenyl)-methylpolysiloxane) capillary column (30 m × 250 µm id and 0.25 µm film thickness). The following oven temperature program was used: 100 °C (1.7 min)–30 °C min−1–210 °C (0 min)–5 °C min−1–300 °C (5 min). The injection volume was 5 µL, helium was used as carrier gas at a flow rate of 3.2 mL/min, nitrogen was used as makeup gas at a flow rate of 60 mL/min and the detector temperature was set to 330 °C.
7
Chia Seed Oil Extraction and Fatty Acid Profile
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The oil extraction was carried out according to the Folch extraction method [25 (link)]. Aliquots of 5 g of ground seeds were weighed and a mixture of chloroform:methanol 2:1 v/v) was added (20:1 v/w). The mixture was stirred and then vacuum filtered to remove the defatted seed meal. Then, the solvent was removed from the filtrate in a rotary evaporator at 40 °C, and the resulting chia oil was kept at −20 °C under an inert nitrogen atmosphere until further analysis.
The fatty acid profile was determined by gas–liquid chromatography coupled with a flame ionization detection (GC-FID) described by Rincón-Cervera et al. [26 (link)] using the Agilent 6890N equipment and a 7683B autosampler (Agilent Technologies, Santa Clara, CA, USA). Fatty acid identification was carried out according to the respective retention times through the capillary column Supelco SP-2560 (100 m × 0.25 mm × 0.2 μm film) (Sigma-Aldrich, St. Louis, MO, USA) compared to analytical standards (37 FAME Mix components from Supelco, Sigma-Aldrich, St. Louis, MO, USA).
The fatty acid profile was determined by gas–liquid chromatography coupled with a flame ionization detection (GC-FID) described by Rincón-Cervera et al. [26 (link)] using the Agilent 6890N equipment and a 7683B autosampler (Agilent Technologies, Santa Clara, CA, USA). Fatty acid identification was carried out according to the respective retention times through the capillary column Supelco SP-2560 (100 m × 0.25 mm × 0.2 μm film) (Sigma-Aldrich, St. Louis, MO, USA) compared to analytical standards (37 FAME Mix components from Supelco, Sigma-Aldrich, St. Louis, MO, USA).
8
Quantifying DCM Residues in Gelatin Scaffolds
9
GC-MS Analysis of Phytochemicals
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Analysis of PAs was carried out on an Agilent GC-MS (CA, USA) consisting of a 7890 GC, a 7683B autosampler and a 5975C MS. The conditions are the same with those in our previous work.31 (link) The GC separation was achieved on an Agilent DB-5MS column (30 m × 0.25 mm × 0.25 μm). The oven temperature was programmed from 100 °C to 210 °C at a rate of 6 °C min−1, then to 300 °C at a rate of 12 °C min−1 and held for 5 min. Helium (99.9995%) was used as the carrier gas at a flow rate of 1.0 mL min−1. The injection volume was 1.0 μL in splitless mode. The temperature of injection port, transfer-line, ion source and quadrupole were held at 300, 300, 230 and 150 °C, respectively. The solvent delay was 8.5 min. The detection was performed in full scan mode with a m/z range from 50 to 550.
10
GC-MS Metabolite Quantification Protocol
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Samples were thawed, diluted with extraction mix to 1.7 mL, and 5 µL IS were added. Aliquots of 0.7 mL per sample were processed as described above. Derivatization was achieved in a reduced volume of 25 µL methoxymation solution and 25 µL silylation solution. Samples with low IS signal were re-derivatized with MSTFA as described above. GC–MS measurements were conducted using a 6890 N GC equipped with a 7683B auto sampler (Agilent) coupled to a Micromass GCT Premier (Waters) mass spectrometer. The GC was operated with glass liners (Agilent, 4 × 6.3 × 78.5 mm inner × outer diameter × length), split 1, and 250 °C injector temperature. The MS was used with 300 °C source temperature and dynamic range extension mode. Resolution was > 6.000 at m/z 501.97.
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