Fresh barberry fruits were prepared from Birjand, Iran. For the preservation of physicochemical properties, the fruits were dried at room temperature. Rapeseed lecithin was provided by an enzymatic extraction approach without applying any organic solvent. Chitosan (shrimps shell source, deacetylation degree up to 75%) was purchased from Sigma-Aldrich (Tokyo, Japan). Other chemicals include acetic acid, hexane, chloroform, ethanol, methanol, BF3/methanol, acetonitrile, 1-(4-Trimethylammonium-phenyl)-6-phenyl-1,3,5-hexatrien (TMA-DPH), sodium citrate, citric acid, potassium chloride, and sodium acetate, which were supplied from Sigma-Aldrich (Paris, France) and Fisher Scientific (Paris, France). All chemicals were of analytical grade.
Bf3 methanol
Manufactured by Merck Group
Sourced in United States, Germany
BF3-methanol is a laboratory reagent used in organic synthesis. It functions as a Lewis acid catalyst, facilitating various chemical reactions. The product specification and technical details are available upon request.
Automatically generated - may contain errors
Lab products found in correlation
Ethanol, by Merck Group (3 mentions)
Methanol, by Merck Group (3 mentions)
Chloroform, by Merck Group (3 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Ascorbic acid, by Merck Group (2 mentions)
Sp 2380, by Merck Group (2 mentions)
Sodium citrate, by Merck Group (1 mentions)
Sodium acetate, by Merck Group (1 mentions)
Citric acid, by Merck Group (1 mentions)
Hexane, by Merck Group (1 mentions)
Chitosan, by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Pufa2, by Merck Group (1 mentions)
Chloroform, by Guangzhou Chemical (1 mentions)
N tetracosane, by Merck Group (1 mentions)
N dotriacontane, by Merck Group (1 mentions)
Hp inowax capillary column, by Agilent Technologies (1 mentions)
Anhydrous sodium sulfate, by Merck Group (1 mentions)
17 protocols using bf3 methanol
1
Barberry Fruit Preservation Protocol
2
Hepatic Lipid and Fatty Acid Profiling
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To assess hepatic TG content, neutral hepatic lipids were extracted in isopropanol and TG measured as previously published [29 (link)]. To assess hepatic fatty acid composition, total lipids were extracted using the Bligh and Dyer method [30 (link)]. An aliquot of extracted lipids was transmethylated with BF3-methanol (Sigma-Aldrich) to quantify specific methyl esters of fatty acids using GC/MS, as we previously reported [31 (link), 32 (link)], using 17 : 1 as the internal standard to quantify the amount of each fatty acid in the sample. In addition, we used a commercial sample of polyunsaturated fatty acid mixture (PUFA-2, Supelco) to identify the different fatty acids in the samples.
3
Cuticular Wax and Cutin Monomer Extraction
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Cuticular waxes and cutin monomers were extracted in series from isolated cuticular membranes according to the previous methods with minor modifications (Huang et al., 2017 (link)). The isolated cuticular membranes were completely dipped in chloroform (Guangzhou Chemical Reagent Factory, China) with a mild temperature of around 40°C to better release the soluble waxes. The extraction time was set for 2 min. Each sample was extracted three times and combined with the extracts. Then, n-tetracosane (Sigma–Aldrich, Shanghai, China) was added as an internal standard to evaluate the cuticular contents. The solvent in the extracts was evaporated by gentle nitrogen gas until they dried for further analysis. After that, the membranes that have been removed of soluble waxes were subsequently depolymerized in boron trifluoride with methanol (BF3-methanol, 10%, ~1.3 M, Sigma–Aldrich, Shanghai, China) and incubated for 16 h at 70°C. After membranes were lysed, n-dotriacontane (Sigma–Aldrich, Shanghai, China) as an internal standard was added. Saturated aqueous sodium chloride solution and chloroform were added in series to extract the cutin monomers. The organic phase was collected and evaporated to dryness under a gentle stream of nitrogen gas for further analysis.
4
Cheese Lipid Extraction and Fatty Acid Analysis
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Forty five milliliter of Folch solution (chloroform:methanol=2:1) was
added to 15 g of the chopped cheese sample by Folch et al. (1957) (link) and homogenized for 10 min. Then, the mixture
was filtered, and following centrifugation (5,000×g, 10°C, 10
min), Na2SO4 was added to the lower layer; after
filtration, chloroform was blown off with a centrifugal concentrator in order to
acquire lipids. Next, 1 mL of 0.5 N NaOH was added to the extracted lipids using
the method described byMorrison and Smith
(1964) . The mixture was then heated at 100°C for 20 min and
cooled. Following the addition of 2 mL of boron trifluoride methanol solution
(BF3 methanol, Sigma, USA), heating, and cooling, 8 mL of NaCl
solution and 1 mL of Heptane were added, and the supernatant was analyzed by gas
chromatography (Varian star 3600, USA). The column of the equipment used for the
analysis was an Omegawax 205 fused-silica bond capillary column with 30
m×0.32 mm dimensions and a 0.25 μm film thickness, as well as a 1
mL/min flow rate of the column. A flame ionization detector was used and
nitrogen gas was applied in order to carry the gas.
added to 15 g of the chopped cheese sample by Folch et al. (1957) (link) and homogenized for 10 min. Then, the mixture
was filtered, and following centrifugation (5,000×g, 10°C, 10
min), Na2SO4 was added to the lower layer; after
filtration, chloroform was blown off with a centrifugal concentrator in order to
acquire lipids. Next, 1 mL of 0.5 N NaOH was added to the extracted lipids using
the method described by
(1964)
cooled. Following the addition of 2 mL of boron trifluoride methanol solution
(BF3 methanol, Sigma, USA), heating, and cooling, 8 mL of NaCl
solution and 1 mL of Heptane were added, and the supernatant was analyzed by gas
chromatography (Varian star 3600, USA). The column of the equipment used for the
analysis was an Omegawax 205 fused-silica bond capillary column with 30
m×0.32 mm dimensions and a 0.25 μm film thickness, as well as a 1
mL/min flow rate of the column. A flame ionization detector was used and
nitrogen gas was applied in order to carry the gas.
5
Cheese Fatty Acid Profiling using GC-MS
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A gas chromatograph (GC) (6890N, Agilent Technologies, Santa Clara, CA, USA) equipped with an Agilent 5975 series mass selective detector was applied to analyze the fatty acid compositions of the cheese samples prepared with palm oil and oleogels. After the cheese was subjected to Soxhlet extraction with diethyl ether [36 (link)], fatty acid methyl ester (FAME) derivatives prepared with KOH methanol and 10% BF3-methanol (Sigma-Aldrich, St. Louis, MO, USA) were separated on an HP-INOWAX capillary column (30 m × 0.32 mm × 0.25 μm, Agilent Technologies), and the flow rate of purified helium carrier gas was 2 mL/min. The injector temperature was 300 °C, and the column temperature was maintained at 100 °C for 5 min that was then heated to 250 °C (3 °C/min) and held for 5 min. The National Institute of Standards and Technology (NIST) 11 mass spectral library (NIST 11) was used for identifying fatty acids in the mass spectral results obtained.
6
Antioxidant Capacity Evaluation Methods
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Folin-Ciocalteu phenol reagent, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ascorbic acid (AA), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), potassium persulfate, 14% boron trifluoride-methanol (BF3-methanol), anhydrous sodium sulfate, n-hexane, sodium hydroxide, fatty acid standards (palmitic, stearic, oleic, linoleic, and linolenic acids), chloroform, ethanol, and methanol were purchased from Sigma Aldrich (St. Louis, MO, USA). CS and FS were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All chemicals were of analytical grade and used without further purification.
7
Lipid Extraction and Fatty Acid Analysis
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Lipids were extracted from cells using the method of Folch-Lees [26 (link)] and from lipid droplets using acetone [23 ]. Individual lipid classes were separated by thin layer chromatography using Silica Gel 60 A plates developed in petroleum ether, ethyl ether, acetic acid (80:20:1) and visualized by rhodamine 6G. Phospholipids and triglycerides, or unesterified fatty acids were scraped from the plates and methylated using BF3 /methanol (Sigma-Aldrich,St. Louis, MO) as described by Morrison and Smith [27 (link)]. The methylated fatty acids were extracted and analyzed by gas chromatography. Gas chromatographic analyses were carried out on an Agilent 7890A gas chromatograph equipped with flame ionization detectors and a capillary column (SP2380, 0.25 mm x 30 m, 0.25 μm film, Sigma-Aldrich). Helium was used as the carrier gas. The oven temperature was programmed from 160°C to 230°C at 4°C/min. Fatty acid methyl esters were identified by comparing the retention times to those of known standards. Inclusion of lipid standards (dipentadecanoyl phosphatidylcholine (C15:0), trieicosenoin (C20:1ω9), and pentadecanoic acid (C15:0)) permitted quantitation of the lipid classes.
8
Fatty Acid Methyl Ester Analysis by GC-FID
9
Metabolome and Azelaic Acid Analysis via GC-MS
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For total metabolome and azelaic acid analysis on GC-MS, a fraction of the dried residual samples were subjected to BF3 methanol derivatization prior to injection as follows: 2 mL of 10% BF3 methanol solution (Sigma, St Louis, MO, USA) was added to approximately 1 mg of the dried sample and incubated in a boiling water bath for 60 min. Subsequently, 2 mL of n-hexane was added and washed with milli-Q water (Millipore) at room temperature. The supernatant solvent was then separated by centrifugation.
10
Lipid Analysis of Thigh Meat
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Lipids were analyzed from thigh meat according to the method of Folch et al.
[16 (link)]. Thigh meat (30 g) was blended
with 150 mL of solution (chloroform : methanol = 2 : 1) and added 0.88%
KOH. The solution was kept at room temperature for 2 hours and the upper phase
was extracted. Then chloroform was evaporated using 99.9% N2 gas and
cooling. 1 mL of BF3-Methanol (Sigma Chemical, St. Louis, MO, USA) was added to
the sample and heated at 70 °C for 30 min. After samples were cooled, 2
mL of Hexan (HPLC grade) and 5 mL of distilled water were added to the solution.
The upper phase is rolled out after vortexing the samples. Fatty acid methyl
ester dissolved in hexane was transferred to a GC vial and the fatty acids were
analyzed using a capillary column (30 mm × 0.32 mm × 0.25
μm film thickness, Omegawax 320, Supelco, Bellefonte, PA, USA) of GC
(Shimadzu Gas Chromatography 17-A, Tokyo, Japan).
The oven temperature was set to 200 °C and helium was divided by a 100:1
split ratio to serce as a carrier gas at linear flow of 0.79 mL/min. Fatty acids
were determined by comparison with standard retention times and the relative
amounts were identicated in weight percent of total fatty acids.
[16 (link)]. Thigh meat (30 g) was blended
with 150 mL of solution (chloroform : methanol = 2 : 1) and added 0.88%
KOH. The solution was kept at room temperature for 2 hours and the upper phase
was extracted. Then chloroform was evaporated using 99.9% N2 gas and
cooling. 1 mL of BF3-Methanol (Sigma Chemical, St. Louis, MO, USA) was added to
the sample and heated at 70 °C for 30 min. After samples were cooled, 2
mL of Hexan (HPLC grade) and 5 mL of distilled water were added to the solution.
The upper phase is rolled out after vortexing the samples. Fatty acid methyl
ester dissolved in hexane was transferred to a GC vial and the fatty acids were
analyzed using a capillary column (30 mm × 0.32 mm × 0.25
μm film thickness, Omegawax 320, Supelco, Bellefonte, PA, USA) of GC
(Shimadzu Gas Chromatography 17-A, Tokyo, Japan).
The oven temperature was set to 200 °C and helium was divided by a 100:1
split ratio to serce as a carrier gas at linear flow of 0.79 mL/min. Fatty acids
were determined by comparison with standard retention times and the relative
amounts were identicated in weight percent of total fatty acids.
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