After grinding the mouse liver tissue with liquid nitrogen, add 3 mL of n-hexane, invert and mix at 50 °C for 30 min, add 3 mL of methanol solution, and derivatize by shaking at 50 °C for 30 min. Add 1 mL of water and mix well. Leave the layers to stand still and take the upper layer for detection, (GC-MS 7890B-5977A, Agilent, Santa Clara, CA, USA).
Gc ms 7890b 5977a
Manufactured by Agilent Technologies
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The GC-MS 7890B-5977A is a gas chromatography-mass spectrometry (GC-MS) system manufactured by Agilent Technologies. It is designed to perform qualitative and quantitative analysis of complex chemical mixtures. The system combines a gas chromatograph for separation of the sample components and a mass spectrometer for the identification and quantification of the separated analytes.
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Lab products found in correlation
Hp 5ms capillary column, by Agilent Technologies (3 mentions)
5500 q trap lc ms ms, by AB Sciex (2 mentions)
Lc 20ad, by Shimadzu (2 mentions)
Fp 528, by Leco (2 mentions)
Db 35 ms ui capillary column, by Agilent Technologies (2 mentions)
Db 5ms column, by Agilent Technologies (2 mentions)
Dvb car pdms, by Merck Group (1 mentions)
Thermo ultimate 3000 system, by Thermo Fisher Scientific (1 mentions)
Thermo q exactive focus, by Thermo Fisher Scientific (1 mentions)
Milli q ultrapure water purification system, by Merck Group (1 mentions)
Hp 5ms column, by Agilent Technologies (1 mentions)
Easy nlc 1200, by Thermo Fisher Scientific (1 mentions)
Q exactive hf x, by Thermo Fisher Scientific (1 mentions)
Q exactive ms, by Thermo Fisher Scientific (1 mentions)
Hp ffap, by Agilent Technologies (1 mentions)
30 protocols using gc ms 7890b 5977a
1
Mouse Liver Lipid Extraction Protocol
2
Volatile Compound Analysis of Persimmon Wines
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HS‐SPME (50/30 µm DVB/CAR/PDMS, Supelco USA) was used to extract the volatile compounds in the inoculated persimmon wine (IPW) and spontaneous persimmon wine (SPW). GC coupled with MS (GC‐MS, 7890B– 5977A, Agilent, USA) was used for separation and identification. A volume of 25 µl of the internal standard (3‐octanol, 300 mg/ml), 1g sodium chloride, and 7.5 µml of the persimmon fermentation were added to a 20‐ml headspace vial. The sample was statically incubated at 40°C for 15 min, followed by a 45‐min extraction of the volatile compounds by an SPME fiber.
The GC conditions were set as follows: Column, HP‐Innowax (60 m × 0.25 mm × 0.25 µm); carrier gas (e.g. He) velocity, 1.4 ml/min; injection temperature, 240°C; splitless mode; oven temperature program at 50°C (maintained 2 min) to 80°C by 3°C/min, then raised to 230°C by 5°C/min; and the final temperature stage retained for 6 min. The MS conditions were set as follows: electron impact (EI) mode at 70 eV, temperature of 230°C, and total ion current scanning range of 33–550 m/z.
The GC conditions were set as follows: Column, HP‐Innowax (60 m × 0.25 mm × 0.25 µm); carrier gas (e.g. He) velocity, 1.4 ml/min; injection temperature, 240°C; splitless mode; oven temperature program at 50°C (maintained 2 min) to 80°C by 3°C/min, then raised to 230°C by 5°C/min; and the final temperature stage retained for 6 min. The MS conditions were set as follows: electron impact (EI) mode at 70 eV, temperature of 230°C, and total ion current scanning range of 33–550 m/z.
3
Fatty Acid Quantification via GC-MS
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Fatty acids were detected using gas chromatography/mass spectrometry (GC–MS 7890B-5977A, Agilent, USA), and the samples were processed as described in our previous experiments [38 (link)].
4
Fatty Acid and Amino Acid Analysis
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Prior to FAA analysis, 80 mg of tissue was homogenized, and 1000 ul pre-treatment solution (acetonitrile: water 1:1) was added. The suspensions were then shaken for 60 min. Next, the samples were centrifuged at 13200 rpm/min for 10 min, and the supernatants were collected. The supernatant fluids were used to determine FAA composition using liquid chromatography-mass spectrometry (Liquid phase: LC-20AD, Shimadzu, Japan; Mass Spectrometry: 5500 Q TRAP LC-MS/MS, AB SCIEX, USA). Pretreatment of fatty acid samples was accomplished by homogenizing 100 mg of tissue. Next, 2 ml of n-hexane was added, and shaken at 50 °C for 30 min, at which point 3 ml of KOH methanol solution (0.4 mol/L) was added. The samples were then shaken for 30 min at 50 °C. Next, 1 ml of water and 2 ml of n-hexane were added, and the samples mixed. The mixture was then allowed to stand for stratification. The upper layer was collected, and fatty acids were detected using gas chromatography-mass spectrometry (GC-MS 7890B-5977A, Agilent, USA).
5
Fatty Acid Composition Analysis Protocol
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The fatty acid composition profiles of diets and fish tissues (liver and muscle) were determined as described by Zuo et al. [45 ] with minor modifications after tests to ensure that all fatty acids were esterified using the following procedures. Briefly, freeze-dried samples (liver samples ~80 mg and muscle samples ~120 mg) were added to a 12 ml volumetric glass tube with a screw top containing a teflon gasket. Three ml 1 M potassium hydroxide in methanol was added and the mixture incubated at 72°C for 20 min in a water bath. After cooling, 3 ml 2 M HCL in methanol was added and the mixture incubated at 72°C for a further 20 min. Finally, 1 ml hexane was added to the mixture, shaken vigorously for 1 min, and then allowed to separate into two layers. Fatty acid methyl esters were separated and measured by GC-MS (Agilent Technologies GC-MS 7890B-5977A, USA) with results presented as percentages of total fatty acids.
6
Fatty Acid Profiling of Crab Feeds
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Fatty acid compositions of feeds and hepatopancreas of swimming crabs were determined essentially as described previously (Luo et al., 2021 (link)). Briefly, around 120 mg of freeze-dried samples were placed into glass tubes and 1 mL of methyl tricosanoate (C23:0) solution (1.0 mg/mL hexane) added as an internal standard for quantification, and total lipid extracted by chloroform/methanol (2:1 by vol.). The extracted total lipids were dried under nitrogen and 3 mL methanolic sulfuric acid added and the samples heated at 80 °C in a water bath for 4 h to prepare fatty acid methyl esters (FAME). Finally, extracted FAME was analyzed by GC–MS using an Agilent Technologies GC–MS (7890B-5977A, USA). The content of each fatty acid was calculated using the following equation:
7
Adipose Tissue Fatty Acid Analysis
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The fatty acid composition test was performed according to the previously reported method (21 (link)). Gas chromatography-mass spectrometry (GC-MS 7890B-5977A, Agilent, United States) was used to detect the fatty acid composition. Then, 100 mg of adipose tissue was homogenized, 2 ml of n-hexane added, and it was shaken for 30 min at 50°C. Next, 3 ml of methanol solution (0.4 moL/L) was added and then shaken for 30 min at 50°C. Lastly, 1 ml of water was mixed with 2 ml of n-hexane and shaken for 20 min at (22 ± 3)°C. Afterward, the solution was allowed to stand for stratification, and the upper layer was separated for gas injection detection. Chromatographic column: DB-23 (30 m × 320 μm × 0.25 μm), the carrier gas was helium, inlet temperature was 250°C; the split ratio was 1/5; injection volume was 1 μl, detector temperature was 230°C, the column oven temperature was 50°C for 1 min, 25°C/min to 175°C, 4°C/min to 230°C for 24.75 min.
8
Amino Acid and Fatty Acid Composition Analysis
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Approximately 30 mg of freeze-dried samples were hydrolyzed in 6 M hydrochloric acid at 110 °C for 24 h. The suspension was diluted with water, and 1 mL of sample was filtered using a 0.2 µm filter [17 (link)]. The samples were analyzed using an HPLC-based automatic amino acid analyzer (Ultimate3000-API 3200 Q TRAP; ThermoFisher Scientific, Waltham, MA, USA), according to the manufacturer’s instructions. The contents of amino acids in the muscle were expressed as mg/g of dried tissue.
Approximately 50 mg of freeze-dried powder samples were mixed with 3 mL of chloroform–methanol–water (volume ratio: 8:4:3). The mixture was shaken for 1 min, followed by an ultrasound in an ice bath for 3 to 5 times, and then centrifuged at 1500 rpm for 15 min. The chloroform layer was transferred to another 10 mL glass centrifuge tube and dried in a vacuum drying oven to obtain fatty acid glyceryl ester. The fatty acid glyceryl ester was analyzed using an automatic fatty acid analyzer (GC-MS 7890B-5977A, Agilent, Santa Clara, CA, USA), as previously described [17 (link)].
Approximately 50 mg of freeze-dried powder samples were mixed with 3 mL of chloroform–methanol–water (volume ratio: 8:4:3). The mixture was shaken for 1 min, followed by an ultrasound in an ice bath for 3 to 5 times, and then centrifuged at 1500 rpm for 15 min. The chloroform layer was transferred to another 10 mL glass centrifuge tube and dried in a vacuum drying oven to obtain fatty acid glyceryl ester. The fatty acid glyceryl ester was analyzed using an automatic fatty acid analyzer (GC-MS 7890B-5977A, Agilent, Santa Clara, CA, USA), as previously described [17 (link)].
9
Meat Quality Traits Evaluation
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The longissimus dorsi and semimembranosus muscles were cut to measure the meat quality traits according to previous reports (3 (link), 16 (link)). Briefly, the pH values were measured at 24 h (pH24h) after slaughter. Meat color measurements cover indicators L* (lightness), a* (redness), and b* (yellowness) after slaughter 24 h (L24h, a24h, and b24h). Drip loss, cooking loss, crude protein, and ash content were determined as described previously (3 (link), 17 (link)). Amino acid profile and fatty acid composition were determined using gas chromatography–mass spectrometry (GC–MS 7890B-5977A, Agilent, Palo Alto, CA, United States) and liquid chromatography–mass spectrometry (Liquid phase was performed on Thermo Ultimate 3,000 system, Thermo Fisher Scientific Inc., Waltham, MA, United States; Mass Spectrometry was performed on Thermo Q Exactive Focus mass spectrometer, Thermo Fisher Scientific Inc., Waltham, MA, United States), respectively.
10
Proximate Composition and Fatty Acid Profiles of Crabs
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Proximate composition and fatty acids. The crude protein, crude lipid, moisture and ash content of diets, muscle and hepatopancreas of the crabs were determined according to the method of the Association of Official Analytical Chemists (26) . The moisture content was determined by drying the samples to a constant weight at 105°C. The crude protein contents (N × 6•25) were assayed by the Dumas combustion method with a protein analyzer (FP-528, LECO). Crude lipid was measured via the petroleum ether extraction method using a Soxtec System HT (SX360, OPSIS), and the ash content was determined after incineration in a muffle furnace at 550°C for 8 h.
Fatty acid compositions of diets, hepatopancreas and muscle were analysed as described in detail previously (27) . In brief, total lipid was extracted with chloroform-methanol (2:1, v/v) and fatty acid methyl esters were then produced from total lipid by methanolic sulphuric acid with butylated hydroxytoluene as an antioxidant. Methyl tricosanoate (23 : 0; Sigma Aldridge Trading Co. Ltd) was used as an internal standard at 1•0 mg/ ml hexane. GC (Agilent Technologies GC-MS 7890B-5977A) was used to analyse fatty acid methyl esters with fatty acids identified by reference to known standards and presented as percentages of area.
Fatty acid compositions of diets, hepatopancreas and muscle were analysed as described in detail previously (27) . In brief, total lipid was extracted with chloroform-methanol (2:1, v/v) and fatty acid methyl esters were then produced from total lipid by methanolic sulphuric acid with butylated hydroxytoluene as an antioxidant. Methyl tricosanoate (23 : 0; Sigma Aldridge Trading Co. Ltd) was used as an internal standard at 1•0 mg/ ml hexane. GC (Agilent Technologies GC-MS 7890B-5977A) was used to analyse fatty acid methyl esters with fatty acids identified by reference to known standards and presented as percentages of area.
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