Determination of carotenoids was performed according to the protocol reported by O’Connell et al. (2007) (link) with modifications as follows. Freeze-dried samples (0.2 g) were saponified in 2 N ethanolic KOH (50 mL) with 10% (w/v) ascorbic acid (10 mL) by boiling in a water bath for 30 min. Saponified samples were then extracted by adding hexane (70 mL) and shaking for 2 min at room temperature. Solvents was removed by evaporation, and residue was re-dissolved in mobile phase solvents [acetronitrile: methanol : dichloromethane (80:11:9, v/v/v) with 0.01% (v/v) triethylamine and 0.01% (w/v) ammonium acetate] (3 mL). Extracts were filtered through 0.2μm PTFE syringe filters and were analyzed by HPLC (Agilent 1100 series HPLC with a photodiode array detector, Agilent Technologies). HPLC was utilized using the VYDAC 201TP54-C18 column (4.6 mm×250 mm, 5μm, Grace, Columbia, MD, USA) with a constant flow rate of 0.7 mL/min at ambient temperature. The presence of carotenoids was visualized at 450 nm. Standards including capsantin, lutein, zeathantin, β-cryptoxanthin, lycopene, α-carotene, trans-β-carotene, and cis-β-carotene (Sigma-Aldrich Co.) were quantitated by comparing retention times and UV spectrum between unknown peaks of sample and authentic standards.
Trans β carotene
Manufactured by Merck Group
Sourced in United States
Trans-β-carotene is a chemical compound that is commonly used in laboratory settings. It is a member of the carotenoid family and is a type of provitamin A. Trans-β-carotene is a natural pigment that can be extracted from various plants and is often used as a colorant or a dietary supplement. In the laboratory, it may be used in various applications, such as research on antioxidant properties or as a reference standard in analytical methods.
Automatically generated - may contain errors
Lab products found in correlation
Lutein, by Merck Group (2 mentions)
α carotene, by Merck Group (2 mentions)
β cryptoxanthin, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Lycopene, by Merck Group (1 mentions)
Agilent 1100 series hplc, by Agilent Technologies (1 mentions)
Zeaxanthin, by Extrasynthese (1 mentions)
Cryptoxanthin, by Merck Group (1 mentions)
Synthetic human plasma, by Merck Group (1 mentions)
Zeaxanthin, by Merck Group (1 mentions)
5 cis lycopene, by CaroteNature (1 mentions)
Nitroblue tetrazolium, by Merck Group (1 mentions)
Absolute ethanol, by Merck Group (1 mentions)
Anhydrous sodium carbonate, by Merck Group (1 mentions)
1 1 diphenyl 2 picrylhydrazyl radical dpph, by Merck Group (1 mentions)
Linoleic acid, by Merck Group (1 mentions)
Folin ciocalteau reagent, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Quercetin, by Merck Group (1 mentions)
α tocopherol, by Merck Group (1 mentions)
6 protocols using trans β carotene
1
Carotenoid Quantification in Foods
2
HPLC-UV-DAD Analysis of Carotenoids
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Carotenoids were extracted by liquid–liquid extraction from plasma samples collected at 0 h and 24 h [32 (link)]. Chromatographic analysis of carotenoids was performed by HPLC-UV-DAD, using an HP 1100 HPLC system (Hewlett-105 Packard, Waldbronn, DE) containing a quaternary pump coupled to a DAD G1315B. The separation was carried out with Milli-Q water, methanol (MeOH) and methyl-tert-butyl ether (MTBE) (Panreac Quimica S.A., Barcelona, Spain), according to a procedure previously validated in our group [32 (link)]. A Waters RP column YMC Carotenoid S-5 µm (250 mm × 4.6 mm) and a precolumn YMC Guard Cartridge Carotenoid S-5 µm (20 mm × 4.0 mαm) were used.
Zeaxanthin (Extrasynthese, Genay, France), lutein, cryptoxanthin, α-carotene, β-carotene 9- and 13-cis-β-carotene (Sigma-Aldrich, St. Louis, MO, USA), lycopene (Fluka, Bucks, Switzerland), and 5-cis-lycopene (CaroteNature GmbH, Münsingen, Switzerland) were used as standards. These were pooled and prepared in synthetic human plasma (Sigma-Aldrich, St. Louis, MO, USA).
The sensitivity of each analyte was 0.703 µmol/L (lutein), 0.352 µmol/L (Zeaxanthin), 0.362 µmol/L (cryptoxanthin), 0.480 µmol/L (trans-β-apo-8’-carotenal), 0.745 µmol/L (13-cis-β-carotene), 0.373 µmol/L (9-cis-β-carotene and trans-β-carotene), and 0.186 µmol/L (trans and cis-lycopenes) [32 (link)].
Zeaxanthin (Extrasynthese, Genay, France), lutein, cryptoxanthin, α-carotene, β-carotene 9- and 13-cis-β-carotene (Sigma-Aldrich, St. Louis, MO, USA), lycopene (Fluka, Bucks, Switzerland), and 5-cis-lycopene (CaroteNature GmbH, Münsingen, Switzerland) were used as standards. These were pooled and prepared in synthetic human plasma (Sigma-Aldrich, St. Louis, MO, USA).
The sensitivity of each analyte was 0.703 µmol/L (lutein), 0.352 µmol/L (Zeaxanthin), 0.362 µmol/L (cryptoxanthin), 0.480 µmol/L (trans-β-apo-8’-carotenal), 0.745 µmol/L (13-cis-β-carotene), 0.373 µmol/L (9-cis-β-carotene and trans-β-carotene), and 0.186 µmol/L (trans and cis-lycopenes) [32 (link)].
3
Antioxidant Activity Assay Protocol
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Analytical grade hydrochloric acid (37%), hexane, chloroform, methanol, absolute ethanol and spectroscopic grade methanol were purchased from Merck, Darmstadt, Germany. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, trans-β-carotene, linoleic acid, tween 40, butylated hydroxytoluene (BHT), allopurinol, quercetin (QN), (+)-α-tocopherol (TOC), Folin-Ciocalteau reagent, phenazine methosulfate (PMS), nitroblue tetrazolium (NBT), β-nicotinamide-adenine dinucleotide (β-NADH, disodium salt), sodium phosphate dibasic 12 hydrate, anhydrous potassium phosphate monobasic, anhydrous sodium carbonate, xanthine and xanthine oxidase (XO) from bovine milk (Grade IV, ammonium sulphate suspension, 0.3unit/mg protein) were all purchased from Sigma Aldrich Chemical Co. (USA).
4
Carotenoid Standards Quantification Protocol
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Methanol, acetonitrile and dichloromethane were purchased from J.T. Baker (Tokyo, Japan). Ethanol, hexane and sodium sulfate were acquired from Neon Analytical Reagents (Suzano, Brazil). Deionized water was made using an Elix water purification system (Millipore, Billerica, MA, USA). The reference standards trans-canthaxanthin (≥95%), trans-astaxanthin (≥97%), β-cryptoxanthin (≥97%), trans-lutein (≥97%), trans-fucoxanthin (≥95%), trans-β-apo-8′-carotene (apocarotenal - ≥ 96%; internal standard), trans-β-carotene (≥95%) and trans-α-carotene (≥95%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Polytetrafluoroethylene sterile membrane (PTFE – 0.22 μm × 25 mm) from Nova Analítica (São Paulo, Brazil).
5
Carotenoid Reference Standard Preparation
6
Development of Carotenoid-Loaded Polymeric Nanocarriers
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Trans-β-carotene (MM 536.87 g/mol, 93% purity) and poly-ε-caprolactone (MM 70.000-90.000 g/mol) were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). The aqueous surfactant Pluronic F68 ® , from BASF Corporation, was kindly donated by Foscher Solutions Provider (London, GB). The oils employed as carriers of the active compound were sunflower oil (Bunge Foods, Santa Catarina, Brazil), caprylic/capric triglycerides (Palm-oil -Malasya) and glycerol (CRQ -Chromate Chemicals Ltda. São Paulo -Brazil). Fatfree, food-grade soybean lecithin with 70% phosphatidylcholine (Lipoid S75) from Lipoid GmbH (Ludwigshafen, Germany) was obtained from Lipid Ingredients & Technologies (São Paulo, Brazil). The surfactant polyethylene glycol sorbitan monolaurate, Tween ® 20 was obtained from Lafan. The lipophilic surfactant sorbitan monooleate, Span™ 80, was obtained from Sigma-Aldrich (St. Louis, MO, USA). Ethyl acetate, acetonitrile, and methanol, all HPLC grade, were obtained from Sigma-Aldrich (St. Louis, MO, USA).
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