The cells used for olefin extraction were collected by centrifugation in glass vials with PTFE screw-caps. The cell pellets were re-suspended in residual supernatant and 100 μl acetic acid was added and thoroughly mixed. One millilitre of methanol and 4 ml of hexane [amended with 10 μg ml−1 triacontane (Tokyo Chemical Industry) as an internal standard] were added. The whole mixture was shaken overnight at 640 rpm in a KS 130B shaker (IKA Werke, Germany). To facilitate phase separation, the samples were centrifuged shortly and the upper hexane phase was used for analysis. GC–MS analysis was performed using a Shimadzu QP2020 system, equipped with a Rxi-5 ms (30 m × 0.25 mm i.d. × 0.25 μm) capillary column. The samples (1 μl) were injected in split mode (split ratio 1:10) and helium flow was maintained at 1 ml min−1. The temperature program of the GC column was 40 °C for 3 min, ramp to 250 °C at 15 °C min−1, and 250 °C for 19 min. The MS was operated under ionization by electron impact at 70 eV and 200 °C; mass spectra were recorded at m/z range of 40–600. Quantification of the olefins by GC–MS was done by direct comparison of their TIC peak areas with the peak area of triacontane of known concentration, which had been added to the hexane used for extraction.
Qp2020 system
Manufactured by Shimadzu
Sourced in Japan
The QP2020 is a gas chromatograph-mass spectrometer (GC-MS) system manufactured by Shimadzu. The system is designed for the qualitative and quantitative analysis of chemical compounds. It provides accurate and sensitive detection and identification of a wide range of substances.
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6 protocols using qp2020 system
1
Quantitative Olefin Extraction and Analysis
2
Quantifying Formaldehyde and 1,3-PDO in GCS Reactions
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Free formaldehyde in GCS reaction mixtures was derivatized with 2,4-dinitrophenylhydrazine (2,4-DNPH) to form 2,4-dinitrophenylhydrazone, before quantified by HPLC (Fig. 2 ). To this end, 0.2 mL of a reaction mixture was mixed with 0.2 mL of 10% (v/v) trifluoroacetic acid (TFA), 0.1 mL of 2,4-DNPH (1 g/L) and 0.5 mL of acetonitrile. Derivatization occurred at 40 °C for 30 min. The 2,4-dinitrophenylhydrazone derivative was analyzed using a Wondasil C18 column (4.6 mm × 150 mm, 5 μm, Shimadzu, Japan) with acetonitrile/water (50,50) containing 0.095% TFA as the mobile phase at a flow rate of 1.0 mL/min, and detected at the wavelength of 352 nm.
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HPLC analysis of formaldehyde from different samples.
3
Quantification of Fecal SCFAs by GC-MS
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Faecal SCFA concentration was determined by GC-MS (Shimadzu QP2020 system with a flame ionization detector), equipped with PAL RTC autosampler (CTC Analytics). Helium was used as the carrier gas and fused silica capillary columns 30 m x 0.25 mm coated with 0.25 µm film thickness were used. The injection port temperature was set to 250 °C. The initial oven temperature was held at 60 °C for 2 min and then raremped to 330 °C at a rate of 15 °C per minute. MS parameters were set to: ion source temperature at 200 °C, interface temperature at 280 °C, and loop time of 0.3 sec. For the GC-MS measurement, 50 µL of faecal samples with a concentration of 0.5 µg/µL and 20 µg/µL prepared in ethanol were mixed with 10 µL of acetic acid-d4 (80 µM). Using PAL RTC autosampler, 4-(4, 6-Dimethoxy-1, 3, 5-triazin-2-yl)-4methylmorpholinium (DMT-MM) and n-octylamine (10 µL of each reagent at a concentration of 80 µM) were added to each faecal samples and reacted for 9 hr prior to injection into GC-MS.
Samples were analysed and quantified using LabSolutions Insight GC-MS software (Shimadzu).
Faecal pH was measured from the supernatant from 0.1 mg/µL of faecal suspension in distilled water using a pH meter (Horiba Ltd.). From the same faecal suspension, faecal ammonia level was quantified using enzymatic ammonia ELISA assay kit (Abcam) according to the manufacture's protocol.
Samples were analysed and quantified using LabSolutions Insight GC-MS software (Shimadzu).
Faecal pH was measured from the supernatant from 0.1 mg/µL of faecal suspension in distilled water using a pH meter (Horiba Ltd.). From the same faecal suspension, faecal ammonia level was quantified using enzymatic ammonia ELISA assay kit (Abcam) according to the manufacture's protocol.
4
Quantification of Formaldehyde and 1,3-PDO
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Free formaldehyde in GCS reaction mixtures was derivatized with 2,4-dinitrophenylhydrazine (2,4-DNPH) to form 2,4-dinitrophenylhydrazone, before quanti ed by HPLC (Fig. 2). To this end, 0.2 mL of a reaction mixture was mixed with 0.2 mL of 10% (v/v) tri uoroacetic acid (TFA), 0.1 mL of 2,4-DNPH (1 g/L) and 0.5 mL of acetonitrile. Derivatization occurred at 40 °C for 30 min. The 2,4-dinitrophenylhydrazone derivative was analyzed using a Wondasil C 18 column (4.6 mm × 150 mm, 5 µm, Shimadzu, Japan) with acetonitrile/water (50:50) containing 0.095% TFA as the mobile phase at a ow rate of 1.0 mL/min, and detected at the wavelength of 352 nm.
1,3-PDO was analyzed by GC-MS as described in Wang et al. using a QP2020 system (Shimadzu, Japan) equipped with a SH-Rxi-5Sil-MS column (Shimadzu, Japan), with helium as the carrier gas. The oven temperature was programmed to be held at 100 °C for 2 min, raised at a gradient of 15 °C min - 1 to 270 °C and held for 12 min at 270 °C.
3 Results And Discussion
1,3-PDO was analyzed by GC-MS as described in Wang et al. using a QP2020 system (Shimadzu, Japan) equipped with a SH-Rxi-5Sil-MS column (Shimadzu, Japan), with helium as the carrier gas. The oven temperature was programmed to be held at 100 °C for 2 min, raised at a gradient of 15 °C min - 1 to 270 °C and held for 12 min at 270 °C.
3 Results And Discussion
5
Quantification of Formaldehyde in Reactions
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Free formaldehyde in GCS reaction mixtures was derivatized with 2,4-dinitrophenylhydrazine (2,4-DNPH) to form 2,4-dinitrophenylhydrazone, before quanti ed by HPLC (Fig. 2). To this end, 0.2 mL of a reaction mixture was mixed with 0.2 mL of 10% (v/v) tri uoroacetic acid (TFA), 0.1 mL of 2,4-DNPH (1g/L) and 0.5 mL of acetonitrile. Derivatization occurred at 40°C for 30 min. The 2,4-dinitrophenylhydrazone derivative was analyzed using a Wondasil C 18 column (4.6 mm×150 mm, 5 µm, Shimadzu, Japan) with acetonitrile/water (50:50) containing 0.095% TFA as the mobile phase at a ow rate of 1.0 mL/min, and detected at the wavelength of 352 nm. 1,3-PDO was analyzed by GC-MS as described in Wang et al. using a QP2020 system (Shimadzu, Japan) equipped with a SH-Rxi-5Sil-MS column (Shimadzu, Japan), with helium as the carrier gas. The oven temperature was programmed to be held at 100°C for 2 min, raised at a gradient of 15°C min -1 to 270°C and held for 12 min at 270°C.
6
Quantification of Formaldehyde and 1,3-PDO
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Free formaldehyde in GCS reaction mixtures was derivatized with 2,4-dinitrophenylhydrazine (2,4-DNPH) to form 2,4-dinitrophenylhydrazone, before quanti ed by HPLC (Fig. 2). To this end, 0.2 mL of a reaction mixture was mixed with 0.2 mL of 10% (v/v) tri uoroacetic acid (TFA), 0.1 mL of 2,4-DNPH (1g/L) and 0.5 mL of acetonitrile. Derivatization occurred at 40°C for 30 min. The 2,4-dinitrophenylhydrazone derivative was analyzed using a Wondasil C 18 column (4.6 mm×150 mm, 5 µm, Shimadzu, Japan) with acetonitrile/water (50:50) containing 0.095% TFA as the mobile phase at a ow rate of 1.0 mL/min, and detected at the wavelength of 352 nm.
1,3-PDO was analyzed by GC-MS as described in Wang et al. using a QP2020 system (Shimadzu, Japan) equipped with a SH-Rxi-5Sil-MS column (Shimadzu, Japan), with helium as the carrier gas. The oven temperature was programmed to be held at 100°C for 2 min, raised at a gradient of 15°C min -1 to 270°C and held for 12 min at 270°C.
1,3-PDO was analyzed by GC-MS as described in Wang et al. using a QP2020 system (Shimadzu, Japan) equipped with a SH-Rxi-5Sil-MS column (Shimadzu, Japan), with helium as the carrier gas. The oven temperature was programmed to be held at 100°C for 2 min, raised at a gradient of 15°C min -1 to 270°C and held for 12 min at 270°C.
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