Ten grams of fermented grains were mixed with 90 mL of distilled water and ultrasonically treated at 0°C for 30 min. The suspension was centrifuged at 2,000 g for 5 min, and the supernatant was pipetted to analyzed the contents of reducing sugar and ethanol. Reducing sugar was analyzed using DNS method (Miller, 1959 (link)). Ethanol content analysis was carried out on an Agilent 7820A GC system equipped with a DB-Wax column (30 m × 0.25 mm × 0.25 μm, J&W Scientific, CA, United States). The oven temperature was held at 60°C for 5 min and then raised to 230°C at a rate of 10°C min–1 before being held at 230°C for 5 min. The carrier gas was helium with a flow-rate of 1 mL min–1. Ethanol contents were quantified by comparing the peak areas with authentic standards. Water content in the fermented grains was measured by drying the samples at 105°C for 24 h. The sampling site temperatures were measured using a thermometer at the time of sampling. Acidity and starch content were assayed using the method described by Wu et al. (2013) (link).
Agilent 7820a gc system
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 7820A GC system is a gas chromatography instrument designed for analytical laboratory applications. It features automated sample injection, temperature-controlled oven, and detector options for separating and analyzing complex chemical mixtures.
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Lab products found in correlation
Db ffap capillary column, by Agilent Technologies (2 mentions)
Db wax column, by Agilent Technologies (1 mentions)
Flame ionization detector, by Agilent Technologies (1 mentions)
Slb il111 capillary column, by Merck Group (1 mentions)
Grain fatty acid methyl ester mix, by Merck Group (1 mentions)
Milli q rg system, by Merck Group (1 mentions)
Fiveeasy f 20 ph meter, by Mettler Toledo (1 mentions)
Hp 5ms quartz capillary column, by Agilent Technologies (1 mentions)
Mikro 220r centrifuge, by Hettich (1 mentions)
12 protocols using agilent 7820a gc system
1
Analysis of Fermented Grains
2
Characterizing Cecum Carbohydrates and Volatile Fatty Acids
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The cellulose content in the cecum was determined by Cellulose Content Kit (Nanjing Jiancheng Bioengineering Institute CO., Ltd., China). The amylose content in the cecum was determined by Megazyme Total Starch Kit (K-TSTA 04/2009, Biostest Co., Ltd., China). The acetate, propionate, butyrate, valerate, isobutyrate, and isovalerate concentrations of hindgut digesta were determined by Agilent 7820A GC system (Agilent, CA, United States) (Li et al., 2014b (link)). About 0.5 g digesta was weighed and diluted with 1 mL ultrapure water. Then, a high-speed homogenizer Scientz-48 (Ningbo Scientz Bioscience Co., Inc., China) was used to make sure the combination was mixed completely. After being centrifuged by centrifugal 5810R (Eppendorf, United States) at 13,500 rpm about 10 min, 1.5 mL supernatant of diluted digesta was moved into 300 μL 25%w/v metaphosphoric solution for further purification. After repeating the previous centrifugation step, 1 mL supernatant was moved into 200 μL 25% crotonic acid, and the mixture was collected into an EP tube passed through a 0.5 μm filter to be measured.
3
Analysis of Phenylacetaldehyde and 2,5-Dimethylpyrazine
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The determination of phenylacetaldehyde and 2,5-dimethylpyrazine levels, which are two other important intermediate compounds of HAs, primarily refers to [31 (link)]. In detail, phenylacetaldehyde extraction involved the mixing of 5 mL of water and 20 mL of ethyl acetate with the sample solutions in separate funnels. Following separation, the ethyl acetate layers underwent filtration before injection into the gas chromatography (GC) system. GC detection was conducted using an Agilent 7820A GC system (Agilent Technologies, Inc., Santa Clara, CA, USA). The headspace autosampler and GC parameters were the same as in our previous study.
Moreover, solid-phase microextraction coupled with gas chromatography-mass spectrometry were used to test 2,5-dimethylpyrazine levels, the injector temperature and the detector temperature, and the flow rates of the fuel gas and oxidant gas were the same as those used in the detection of phenylacetaldehyde.
Moreover, solid-phase microextraction coupled with gas chromatography-mass spectrometry were used to test 2,5-dimethylpyrazine levels, the injector temperature and the detector temperature, and the flow rates of the fuel gas and oxidant gas were the same as those used in the detection of phenylacetaldehyde.
4
SCFA Quantification by GC-FID
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The SCFA contents in the supernatant were detected by a GC process. The liquid sample was centrifuged at 12 000 rpm for 1 min, and 600 μL of the supernatant was acidized with 200 μL 20% H2SO4 (v/v) and vortexed adequately for 1 min. Then, 500 μL n-butanol was added to the acidized fluid for liquid–liquid extraction, and the mixture was vortexed adequately for 2 min and centrifuged at 10 000 rpm for 1 min. Finally, the upper organic phase was filtered through a 0.22 μm filter membrane for further GC detection.
An Agilent 7820A GC system (Agilent Technologies, Santa Clara, CA, USA) was equipped with a flame ionization detector (FID) and an automatic sampler, a DB-FFAP capillary column (Agilent, 30 m × 0.25 mm × 0.25 μm) was used for component separation. The temperature operating conditions of the GC system are as follows: initial column temperature, 80 °C, held for 1 min; then 80 °C to 160 °C at a rate of 3 °C min−1; and finally, 160 °C to 230 °C, held for 1 min. Other parameters were set as follows: injection port temperature, 300 °C; FID temperature, 320 °C; and injection volume, 1 μL. The flow rates of dry air, hydrogen and nitrogen were 300, 30, and 20 mL min−1, respectively. Each experiment was performed in triplicate.
An Agilent 7820A GC system (Agilent Technologies, Santa Clara, CA, USA) was equipped with a flame ionization detector (FID) and an automatic sampler, a DB-FFAP capillary column (Agilent, 30 m × 0.25 mm × 0.25 μm) was used for component separation. The temperature operating conditions of the GC system are as follows: initial column temperature, 80 °C, held for 1 min; then 80 °C to 160 °C at a rate of 3 °C min−1; and finally, 160 °C to 230 °C, held for 1 min. Other parameters were set as follows: injection port temperature, 300 °C; FID temperature, 320 °C; and injection volume, 1 μL. The flow rates of dry air, hydrogen and nitrogen were 300, 30, and 20 mL min−1, respectively. Each experiment was performed in triplicate.
5
Gas Chromatography Analysis of Organic Compounds
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An Agilent
7820A GC system (Agilent Technologies, Santa Clara, CA) was equipped
with a flame ionization detector (FID), and a DB-FFAP capillary column
(Agilent, 30 m × 0.25 mm × 0.25 μm) was used for component
separation. The operating temperature conditions of the GC system
were as follows: initial column temperature was 80 °C (held for
1 min), then increased from 80 to 160 °C at a rate of 3 °C/min,
and finally increased from 160 to 230 °C, which was held for
1 min. Other parameters were set as follows: injection port temperature,
300 °C; FID temperature, 320 °C; and injection volume, 1
μL. The flow rates of dry air, hydrogen, and nitrogen were 300,
30, and 20 mL/min, respectively. Each experiment was performed in
triplicate.
7820A GC system (Agilent Technologies, Santa Clara, CA) was equipped
with a flame ionization detector (FID), and a DB-FFAP capillary column
(Agilent, 30 m × 0.25 mm × 0.25 μm) was used for component
separation. The operating temperature conditions of the GC system
were as follows: initial column temperature was 80 °C (held for
1 min), then increased from 80 to 160 °C at a rate of 3 °C/min,
and finally increased from 160 to 230 °C, which was held for
1 min. Other parameters were set as follows: injection port temperature,
300 °C; FID temperature, 320 °C; and injection volume, 1
μL. The flow rates of dry air, hydrogen, and nitrogen were 300,
30, and 20 mL/min, respectively. Each experiment was performed in
triplicate.
6
Analysis of Phenylacetaldehyde and 2,5-Dimethylpyrazine
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The determination of phenylacetaldehyde and 2,5-dimethylpyrazine levels, which are two other important intermediate compounds of HAs, primarily refers to [31 (link)]. In detail, phenylacetaldehyde extraction involved the mixing of 5 mL of water and 20 mL of ethyl acetate with the sample solutions in separate funnels. Following separation, the ethyl acetate layers underwent filtration before injection into the gas chromatography (GC) system. GC detection was conducted using an Agilent 7820A GC system (Agilent Technologies, Inc., Santa Clara, CA, USA). The headspace autosampler and GC parameters were the same as in our previous study.
Moreover, solid-phase microextraction coupled with gas chromatography-mass spectrometry were used to test 2,5-dimethylpyrazine levels, the injector temperature and the detector temperature, and the flow rates of the fuel gas and oxidant gas were the same as those used in the detection of phenylacetaldehyde.
Moreover, solid-phase microextraction coupled with gas chromatography-mass spectrometry were used to test 2,5-dimethylpyrazine levels, the injector temperature and the detector temperature, and the flow rates of the fuel gas and oxidant gas were the same as those used in the detection of phenylacetaldehyde.
7
Fatty Acid Profiling of Bread Samples
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Fatty acid profile was obtained by gas chromatography (Agilent 7820A GC System equipped with Flame Ionization Detector, Agilent Technologies, Inc., Santa Clara, CA, USA) according to the ISO 12966-2:2017. Analysis was carried out in duplicate for two samples of each bread.
8
Quantifying Volatile Fatty Acids in Biological Samples
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For the VFA measurements, the rumen fluid and mouse intestinal contents (dissolve each 0.5 g in 1 mL sterile water until completely dissolved) were centrifuged at 13,000 × g for 10 min, and then detected as previously described8 (link). In brief, each 4 mL supernatant was mixed with 1 mL metaphosphoric acid (250 g/L), then centrifuged for 15 min at 10,000 × g at 4 °C. Two milliliters of the supernatant was mixed with 200 µL of crotonic acid (10 g/L) and then filtered through a 0.45 µm filter. The VFAs were separated and quantified with an Agilent 7820 A GC system equipped with a polar capillary column (AE-FFAP, 30 m × 0.25 mm × 0.33 μm) and a flame ionization detector (FID).
9
Fatty Acid Composition Analysis by GC-FID
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The fatty acid composition was determined according to AOCS Official Method Ce 1h-05 [45 ]. Briefly, oil was dissolved in hexane and transesterified with sodium methoxide. Fatty acid methyl esters (FAME) were analyzed using an Agilent 7820A GC system (Agilent Technologies, Santa Clara, CA, USA) equipped with a SLB-IL111 capillary column (100 m, 0.25 mm, 0.20 μm, Supelco, Bellefonte, PA, USA) and a flame ionization detector (FID). The FAME mixture was separated under the following conditions: the initial oven temperature was 150 °C, and it increased to 200 °C at 1.5 °C/min; the injector and detector temperature were 250 °C; split 1:10; the helium flow rate was 1 mL/min. The FAME were identified by comparison with commercially available standards–grain fatty acid methyl ester mix (Supelco). The results were expressed as the percentage of total fatty acids (see Supplementary Materials ).
10
Lipid and Fatty Acid Analysis of CA and IC
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Lipid quantification in CA and IC was performed by Soxhlet extraction as described in AOAC Official Method 945.16. using petroleum ether. Results were obtained by weighing the dry product containing the lipid fraction and were expressed as % dry matter.
Fatty acid profile was determined by gas chromatography according to ISO 12966-2:2017; using a flame ionization detector (Agilent 7820A GC system, Agilent Technologies, Inc., Santa Clara, CA, USA) [22 (link)]. Analysis was carried out by the Analysis Services Unit facilities of the Institute of Food Science, Technology and Nutrition (ICTAN, CSIC, Madrid, Spain). Analysis was carried out in duplicate and results were expressed as grams per 100 g of sample.
Both determinations were performed as previously described [23 (link)].
Fatty acid profile was determined by gas chromatography according to ISO 12966-2:2017; using a flame ionization detector (Agilent 7820A GC system, Agilent Technologies, Inc., Santa Clara, CA, USA) [22 (link)]. Analysis was carried out by the Analysis Services Unit facilities of the Institute of Food Science, Technology and Nutrition (ICTAN, CSIC, Madrid, Spain). Analysis was carried out in duplicate and results were expressed as grams per 100 g of sample.
Both determinations were performed as previously described [23 (link)].
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