Cholesterol standard (purity 98%), the internal standard 5α-cholestan-3β-ol (purity 98%), pyridine, bis(trimethylsilyl) trifluoroacetamide (BSTFA) with 1% of trimethylchlorosilane (TMCS) and potassium hydroxide were purchased from Sigma-Aldrich (Arklow, Co., Wicklow, Ireland). Chloroform was purchased from Sigma-Aldrich (Oslo, Norway). Methanol, hexane and dichloromethane were purchased from Fisher Scientific (Dublin, Ireland). Ultra-pure water (18.2 MΩ cm−1) was generated in-house using a Millipore water purification system (Millipore, Cork, Ireland). All chemicals were GC grade.
5α cholestan 3β ol
Manufactured by Merck Group
Sourced in United States, Ireland, Germany
5α-cholestan-3β-ol is a chemical compound used in laboratory research and analysis. It is a steroid alcohol derived from cholesterol. This product serves as a reference standard or analytical tool for various applications in the life sciences and chemical fields.
Automatically generated - may contain errors
Lab products found in correlation
Trimethylchlorosilane tmcs, by Merck Group (2 mentions)
Bis trimethylsilyl trifluoroacetamide bstfa, by Merck Group (2 mentions)
Zb 5ht inferno capillary column, by Phenomenex (2 mentions)
Methanol, by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
Campesterol, by Merck Group (2 mentions)
β sitosterol, by Merck Group (2 mentions)
β carotene, by Merck Group (2 mentions)
Squalene, by Merck Group (2 mentions)
Lanosterol, by Merck Group (2 mentions)
24α ethyl cholesterol, by Merck Group (2 mentions)
24α methyl cholesterol, by Merck Group (2 mentions)
Hexane, by Thermo Fisher Scientific (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
Chloroform, by Merck Group (1 mentions)
Potassium hydroxide, by Merck Group (1 mentions)
Dichloromethane, by Thermo Fisher Scientific (1 mentions)
Pyridine, by Merck Group (1 mentions)
Millipore water purification system, by Merck Group (1 mentions)
N hexane, by Merck Group (1 mentions)
14 protocols using 5α cholestan 3β ol
1
Cholesterol Quantification by GC-MS
2
Quantification of Olive Oil Sterols and Squalene
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Around 200 mg of each sampled oil was placed in a 10 mL propylene tube. Then, 200 μL of an internal standard solution (5α-cholestan-3β-ol, Sigma-Aldrich) in hexane was added. The analysis of the unsaponifiable fraction was performed by GC, without a preliminary thin-layer chromatography fractionation. This approach was used for the analysis of sterols and squalene in olive fruit and olive oil by many authors [30 (link),31 (link)]. Alkaline hydrolysis was performed by adding 2 mL of KOH 2%, then tubes were soaked in a water bath at 80 °C for 15 min, and the unsaponifiable fraction was extracted by vortexing with 1 mL hexane and 1.5 mL NaCl 1%. The upper hexane layer was transferred to 2 mL glass vials. Samples were conserved at −20 °C until analysis, within 24 h from preparation.
Analyses were performed on a GC-FID, using a ZB-5HT Inferno capillary column (15 m × 0.32 mm × 0.10 μm film thickness, Phenomenex, Torrance, CA, USA). The carrier gas was helium (column flow 1.5 mL/min), and the split ratio was 1:100. The oven temperature was programmed as follows: 0.5 min at 150 °C, from 150 °C to 240 °C at 8 °C/min and from 240 °C to 370 °C at 25 °C/min held for 5 min, at 370 °C, followed by 320 °C for the injector and 350 °C for the detector (FID). The quantification was performed by external standards of squalene and sterols purchased from Sigma-Aldrich (St. Louis, MO, USA).
Analyses were performed on a GC-FID, using a ZB-5HT Inferno capillary column (15 m × 0.32 mm × 0.10 μm film thickness, Phenomenex, Torrance, CA, USA). The carrier gas was helium (column flow 1.5 mL/min), and the split ratio was 1:100. The oven temperature was programmed as follows: 0.5 min at 150 °C, from 150 °C to 240 °C at 8 °C/min and from 240 °C to 370 °C at 25 °C/min held for 5 min, at 370 °C, followed by 320 °C for the injector and 350 °C for the detector (FID). The quantification was performed by external standards of squalene and sterols purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Olive Pulp Unsaponifiable Fraction Extraction
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Around 200 mg of ground olive pulp was placed in a 10 mL propylene tube. Two hundred microliters of an internal standard solution (5α-cholestan-3β-ol, Sigma-Aldrich) in hexane was added. The analysis of the unsaponifiable fraction by GC was previously reported in [6 (link),59 (link),63 (link)].
4
Fatty Acid Profiling via GC-MS
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A certified fatty acids methyl ester (FAME) reference standard mixture (37 fatty acids from C4 to C24) was obtained from Sigma-Aldrich (St. Louis, MO, USA). The internal standard 5α-Cholestan-3β-ol (purity ≥ 95%) was purchased from Sigma-Aldrich. The other reagents were supplied by Sinopharm (Shanghai, China). The solvents, n-hexane, methanol and acetonitrile, were of HPLC grade (Sigma-Aldrich). The analytical standards, including benzoic acid, cinnamic acid, p-hydroxyphenylacetic acid, naringin, paeoniflorin, paeonol and oxyresveratrol, were purchased from Aladdin Co., Ltd. (Shanghai, China).
5
Extraction and Analysis of Sea Buckthorn Oil
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Crude SBO was extracted from frozen dried sea-buckthorn pulp by cold pressing. It was provided by Aikang Sea Buckthorn Pharmaceutical Co., Ltd (Xian, Shanxi, China). The neutralized, bleached and deodorized SBOs were prepared in laboratory, and the detailed procedures are listed in Section 2.2.
Standards of 5α-cholestan-3β-ol, tocopherols (α-, β-, γ- and δ-tocopherols), phytosterols (campesterol, β-sitosterol, and 7-stigmasterol), and β-carotene were purchased from Sigma-Aldrich (Shanghai, China). 3-Monochloropropane-1,2-diol (3-MCPD) and 3-MCPD-d5 were purchased from CDN Isotopes Inc (Pointe-Claire, Canada). A standard mixture of PAHs was purchased from AccuStandard (New Haven, USA). BSTFA + TMCS (99 : 1) for derivatization was obtained from Sigma-Aldrich (Shanghai, China). The HPLC grade solvents, such as tetrahydrofuran, acetone, n-hexane, toluene, isopropanol, dichloromethane, acetonitrile, chloroform, were provided by CNW (Darmstadt, Germany). All other reagents and solvents were obtained from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China).
Standards of 5α-cholestan-3β-ol, tocopherols (α-, β-, γ- and δ-tocopherols), phytosterols (campesterol, β-sitosterol, and 7-stigmasterol), and β-carotene were purchased from Sigma-Aldrich (Shanghai, China). 3-Monochloropropane-1,2-diol (3-MCPD) and 3-MCPD-d5 were purchased from CDN Isotopes Inc (Pointe-Claire, Canada). A standard mixture of PAHs was purchased from AccuStandard (New Haven, USA). BSTFA + TMCS (99 : 1) for derivatization was obtained from Sigma-Aldrich (Shanghai, China). The HPLC grade solvents, such as tetrahydrofuran, acetone, n-hexane, toluene, isopropanol, dichloromethane, acetonitrile, chloroform, were provided by CNW (Darmstadt, Germany). All other reagents and solvents were obtained from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China).
6
Sterol Extraction and Identification
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All reagents were purchased from Merck (Darmstadt, Germany). GLC-463 mix was from Nu-Check (Elysian, MN, USA), while FAME 189-19, (+)-α-tocopherol, 5α-cholestan-3β-ol, campesterol, campestanol, stigmasterol, β-sitosterol, sitostanol and Δ5-avenasterol were from Sigma-Aldrich (St. Louis, MO, USA).
7
Fatty Acid Methyl Esters Characterization
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A mixture of standard fatty acid methyl esters (FAMEs), standards of syringic, tyrosol, and 5α-cholestan-3β-ol were sourced from Sigma-Aldrich (Steinheim, Germany). Diethyl ether, acetic acid (glacial), chloroform, and methanol were obtained from ISOLAB (Eschau, Germany). Sodium hydroxide, sodium carbonate, sodium thiosulfate, phosphoric acid, and ethanol were purchased from Fluka (Buchs, Switzerland). p-Anisidine, cyclohexane, and acetonitrile were obtained from Merck (Darmstadt, Germany).
8
Pomegranate Preservation and Analysis
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Commercially mature pomegranate fruits (cv. Wonderful) were procured from Sonia pack-house (33°34′851″ S, 19°00′360″ E) in Western Cape Province, South Africa. The fruits were stored in the Postharvest Technology Research Laboratory at Stellenbosch University at 7.5 ± 0.5 °C and 92% ± 3% RH prior to processing to preserve quality [59 (link)]. The chemical reagents used in the current study including Folin–Ciocalteau reagent, gallic acid, β-Carotene, heptadecanoic acid, 2,4,6-Tri(2-pyridyl)-s-triazine (TPTZ), 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), 5α-Cholestan-3β-ol and 2,2-Diphenyl-1-picrylhydrazyl (DPPH), were supplied by Sigma, South Africa. All the other solvents and reagents were of analytical grade.
9
Cholesterol Quantification Protocol
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Cholesterol standard (purity 98%), the internal standard (ISTD) 5α-cholestan-3β-ol (purity 98%), bis (trimethylsilyl) trifluoroacetamide (BSTFA) with 1% of trimethylchlorosilane (TMCS), heptane, dichloromethane (DCM) and methanol (all GC grade) were sourced from Sigma-Aldrich Co. (Arklow, Co. Wicklow, Ireland). Pyridine was supplied by Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Deionised water (DI) was sourced from a Millipore Elix 15 water purification system (Merck Millipore, Darmstadt, Germany).
10
Camellia Oils: Comprehensive Characterization
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Seventeen camellia oils were collected from different factories in 2017. Among which, nine C. oleifera seed oils and three C. sinensis seed oils were purchased from the local market in Ganzhou city, Jiangxi Province of China; three C. japonica seed oils, two C. sinensis seed oils, extra virgin olive oil, and rapeseed oil were purchased from Tokyo in Japan. Detailed information describing the oil samples is shown in Table 1, which includes the brands, place of production, processing mode, and date of expiry. All the oil samples were stored at -20℃ in a freezer to prevent oxidation before analysis.
Boron trifluoride, tocopherol standard, lanosterol, squalene and 5α-cholestan-3β-ol were purchased from Sigma-Aldrich (St. Louis, MO, USA) . The other reagents were purchased from Wako Pure Chemical Co. (Osaka, Japan) . All chemicals were of analytical reagent grade, and acetone, acetonitrile, n-hexane and 2-propanol were HPLC grade.
Boron trifluoride, tocopherol standard, lanosterol, squalene and 5α-cholestan-3β-ol were purchased from Sigma-Aldrich (St. Louis, MO, USA) . The other reagents were purchased from Wako Pure Chemical Co. (Osaka, Japan) . All chemicals were of analytical reagent grade, and acetone, acetonitrile, n-hexane and 2-propanol were HPLC grade.
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