All digestive enzymes (α-amylase from porcine pancreas, pepsin from hog stomach, pancreatin from porcine pancreas, and bile extract porcine), magnesium hydroxide carbonate, calcium chloride dehydrate, magnesium chloride hexahydrate (99%), magnesium sulphate hexahydrate, sodium chloride, sodium bicarbonate and sodium phosphate were purchased from Sigma-Aldrich (St Louis, MO, USA). Potassium chloride was obtained from Panreac (Barcelona, Spain). Monopotassium phosphate was purchased from Acros Organics (New Jersey, USA). Butyl hydroxytoluene (BHT), hydrochloric acid and sodium hydroxide were acquired from Scharlau Chemie S.A. (Barcelona, Spain). Lycopene, γ-carotene, δ-carotene, β-carotene, lutein, phytofluene and phytoene were obtained from Carote-Nature (Ostermundigen, Switzerland).
Lutein
Manufactured by CaroteNature
Sourced in Switzerland
Lutein is a natural carotenoid compound found in plants. It is commonly extracted and purified for use in various laboratory applications, including analytical testing and research. Lutein exhibits specific absorbance and fluorescence properties that make it useful as a standard or reference material in various analytical techniques.
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Lab products found in correlation
β carotene, by CaroteNature (12 mentions)
Zeaxanthin, by CaroteNature (9 mentions)
α carotene, by CaroteNature (6 mentions)
Lycopene, by CaroteNature (5 mentions)
β cryptoxanthin, by CaroteNature (5 mentions)
Violaxanthin, by CaroteNature (4 mentions)
Acetic acid, by Merck Group (3 mentions)
Neoxanthin, by CaroteNature (3 mentions)
Antheraxanthin, by CaroteNature (3 mentions)
Butylated hydroxytoluene, by Merck Group (3 mentions)
β apo 8 carotenal, by CaroteNature (3 mentions)
Phytoene, by CaroteNature (2 mentions)
Phytofluene, by CaroteNature (2 mentions)
δ carotene, by CaroteNature (2 mentions)
γ carotene, by CaroteNature (2 mentions)
Pepsin, by Merck Group (2 mentions)
Gallic acid, by Merck Group (2 mentions)
Folin ciocalteu reagent, by Merck Group (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions)
Sodium carbonate, by Merck Group (2 mentions)
20 protocols using lutein
1
Comprehensive Gastrointestinal Enzyme Analysis
2
Carotenoid Quantification in Samples
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The carotenoid standards were lutein, zeaxantin, violaxantin, and β-carotene (CaroteNature, Münsingen, Switzerland). Gallic acid (842649), pepsin (P7000; ≥250 U/mg), pancreatin (P7545; 8xUSP), invertase (I4504; ≥300 U/mg), amyloglucosidase (A7095; ≥260 U/mL), Folin–Ciocalteu phenol reagent (109001), nitric acid 60% ultrapure (101518), and chemicals, at analytical or LiChrosolv grade, were purchased from Merck KGaA (Darmstadt, Germany).
3
Carotenoid Extraction and Analysis
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The carotenoid extraction and saponification processes were performed under the condition of weak or dark light to avoid decomposition. The carotenoid pigments were analyzed by reversed phase high-performance liquid chromatography (HPLC) using the modified binary gradient elution procedure originally developed by Lee et al. (2001) (link). The forms of the carotenoids were identified by the retention time of the standard sample and the characteristic spectral value. Standard samples (β-cryptoxanthin, violaxanthin, and lutein) were purchased from CaroteNature (Lupsingen, Switzerland).
4
Characterization of Antioxidant Compounds
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Carbon dioxide (Industrial grade) was obtained from Praxair S.A. (Guadalajara, Mexico) and ethanol from Reactivos Guadalajara (Guadalajara, Mexico). The α-tocopherol, thiamine, riboflavin, niacin, pyridoxine, ammonium acetate, acetic acid, fatty acid methyl esters mix (Supelco 37 component FAME Mix), Folin–Ciocalteu reagent, sodium carbonate, DPPH, sodium acetate, TPTZ, ferric chloride hexahydrate, trolox, and ABTS were obtained from Sigma-Aldrich (Toluca, Mexico). Lutein and β-carotene (analytical grade) were purchased from CaroteNature (Lupsingen, Switzerland). Methanol, acetone, hexane, sulfuric acid and water (HPLC grade) were acquired from Tedia (Monterrey, Mexico). Helium and nitrogen were purchased from Infra S.A. (Monterrey, Mexico). All reagents were ACS grade, unless otherwise stated.
5
Carotenoid Standards Acquisition and Purification
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Violaxanthin, neoxanthin, antheraxanthin, lutein, zeaxanthin, phytoene, β-cryptoxanthin, phytofluene, α-carotene, β-carotene, ζ-carotene, δ-carotene, γ-carotene, neurosporene and lycopene were purchased from CaroteNature (Lupsingen, Switzerland). Methanol was purchased from Fisher Scientific (Waltham, MA, USA). Tert-methyl butyl ether (MTBE), chloroform and dichloromethane were purchased from Avantor Performance Materials (Panoli, Gujrat, India), hexane from Sigma chemical Co. (St. Louis, MO, USA) and ethanol from Hayman Ltd. (Essex, USA). All reagents were HPLC grade or higher. A Millipore Milli-Q water purification system was used to obtain high purity of water.
6
Quantification of Bioactive Compounds
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Reagents were purchased from various suppliers as follows: Authentic standards of kuromanin (cyanidin 3-O-glucoside chloride), chlorogenic acid (3-caffeoylquinic acid), gentisic acid, rutin (quercetin 3-O-rutinoside) (Extrasynthèse, Genay, France); the standard for carotenoids (lutein, carotene, α-carotene, β-carotene and lycopene) were purchased from CaroteNature (Lupsingen, Switzerland); gallic acid, Folin-Ciocalteu's phenol reagent, Trolox [(S)-(-)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid], ABTS [2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)], fluorescein disodium, AAPH [2,2′-azobis (2-methyl-propionamide)], ascorbic acid, meta-phosphoric acid, DTT (dithiothreitol), BHT (butylated hydroxytoluene) as well as acetonitrile, ethanol, methanol, acetone, methyl tert-butyl ether, formic acid and acetic acid (all HPLC grade) (Sigma-Aldrich, St. Louis, MO, USA). In all experiments Milli-Q (Merck Millipore, Darmstadt, Germany) water was used.
7
Detailed Methodology for Phytochemical Analysis
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Methanol, formic acid, isopropanol, n-hexane, ethanol, methyl tert-butyl ether, butylated hydroxytoluene (BHT), HPLC-grade acetonitrile, and acetone were purchased from Fisher (Pittsburgh, PA, USA), all of which were HPLC-grade. Whereas, an Elix water purification system (Millipore, Billerica, MA, USA) was utilized to prepare in-house deionized water. The reference standards lutein (>96 % purity) and β-carotene (>99 % purity) were acquired from Carote Nature (Lupsingen, Switzerland) and measured by HPLC–PDA. Chlorophyll a (>92 % purity) and chlorophyll b (>92 % purity) were obtained from Sigma-Aldrich (St. Louis, MO, USA), and also measured by HPLC–PDA. The standards of delphinidin-3-O-glucoside (≥95 % purity), cyanidin (≥98 % purity), cyanidian-3,5-O-diglucoside (≥97 % purity), delphinidin (≥98 % purity), and cyanidin 3-O-galactoside (≥98 % purity) were obtained from Extrasynthese (Genay, France). All standards were of HPLC grade.
8
Carotenoid Standards Synthesis and Characterization
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α-Carotene, β-carotene, β-cryptoxanthin, lutein, zeaxanthin, antheraxanthin, and lutein-5,6-epoxide were purchased from Carote Nature GmbH (Münsingen, Switzerland). Zeinoxanthin used for standard was synthesized by Wittig condensation of C25-apocarotenal derived from commercially available α-ionone, with previously prepared C15-phosphonium salt possessing 3-hyxroxy-β-end group [33 (link)]. 1H NMR data of synthetic zeinoxanthin were identical with those reported in Reference [34 (link)].
9
Carotenoids and Tocopherols Extraction
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Solvents were commercially obtained (Sigma Aldrich, St. Luis, MO, USA), at the highest quality, and used without further purification. All other used reagents were of analytical grade. Lutein was purchased from CaroteNature (Lupsingen, Switzerland); all-trans-β-carotene was from Sigma Chemicals (St. Luis, MO, USA). α-, β-, γ- and δ-Tocopherol standards were from Merck (Darmstadt, Germany); α-, β-, γ- and δ-Tocotrienol standards were purified, as reported in Panfili et al. [25 (link)]. Purity for all standards was above 95% (as certified by the suppliers).
10
Antioxidant Capacity Assessment Protocol
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Ammoniac (NH3), ammonium acetate, boric acid, ferulic acid, hydrochloric acid (HCl), kaempferol-3-O-glucoside, methanol, natrium hydroxide (NaOH), quercetin-3-O-glucoside, sinapic acid, and tert-butyl methyl ether were obtained from Carl Roth GmbH + Co. KG (Karlsruhe, Germany). Acetic acid, 2-propanol, 2,2-diphenyl-1-picrylhydrazyl (DPPH), methylene chloride, potassium persulfate, and Trolox® (6-hydroxyl-2,5,7,8-tetramethylchroman-2carbonic acid) were purchased from Merck KGaA (Darmstadt, Germany). 2,2′-Azinobis-(3-ethylbenzthiazoline-6sulfonic acid) (ABTS) was purchased from Roche Diagnostic GmbH (Mannheim, Germany) and sulfuric acid from Grüssing GmbH (Filsum, Germany). Acetonitrile was obtained from J.T. Baker (Fisher Scientific GmbH, Grießheim, Germany) and tetrahydrofurane was from VWR International GmbH (Darmstadt, Germany). The standards of β-carotene, chlorophyll a, chlorophyll b, lutein, and zeaxanthin were purchased from CaroteNature® GmbH (Münsingen, Switzerland) and pheophytin a from LGC Standards GmbH (Wesel, Germany).
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