Acetonitrile and methanol were of HPLC grade and purchased from Roth GmbH (Karlsruhe, Germany). Reference compounds (crocin, safranal, apigenin, rutin, caffeic acid, ferulic acid, chlorogenic acid, gallic acid) were purchased from ChromaDex (Santa Ana, CA), Sigma-Aldrich (Saint Louis, MO), HWI Analytik GmbH, and Roth GmbH (Karlsruhe, Germany).
Rutin
Manufactured by ChromaDex
Sourced in United States, Germany
Rutin is a natural plant compound that can be extracted and used as a laboratory reagent. It functions as an antioxidant and has the chemical formula C27H30O16.
Automatically generated - may contain errors
Lab products found in correlation
Quercetin, by ChromaDex (5 mentions)
Chlorogenic acid, by ChromaDex (3 mentions)
Kaempferol, by ChromaDex (3 mentions)
Gallic acid, by ChromaDex (2 mentions)
Quercetin, by Merck Group (2 mentions)
Ellagic acid, by ChromaDex (2 mentions)
Trifluoroacetic acid tfa, by Merck Group (2 mentions)
Myricetin, by ChromaDex (2 mentions)
Ferulic acid, by ChromaDex (1 mentions)
Caffeic acid, by ChromaDex (1 mentions)
Methanol, by Carl Roth (1 mentions)
Acetonitrile, by Carl Roth (1 mentions)
Apigenin, by ChromaDex (1 mentions)
Hplc grade methanol, by Avantor (1 mentions)
Acetic acid, by Avantor (1 mentions)
Isopropanol, by Avantor (1 mentions)
Acetonitrile, by Avantor (1 mentions)
Tetrahydrofuran, by Avantor (1 mentions)
Hplc grade phosphoric acid, by Avantor (1 mentions)
Lachrom elite, by Hitachi (1 mentions)
9 protocols using rutin
1
Analytical Standards for Phytochemicals
2
HPLC Analysis of Phenolic Compounds
3
HPLC-DAD Analysis of Phytochemicals
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HPLC-DAD analysis was conducted using the chromatograph LaChrom Elite (Hitachi, Tokyo, Japan) equipped with an L2455 DAD detector adjusted in the range of 230 to 400 nm, L2200 injector, L2130 pump, L2300 column oven, and a C18 column (150 × 4.6 mm, 5 µm, Merck, Darmstadt, Germany). Data were obtained with EZChrom Elite software, version 3.3.2 SP1 (Santa Clara, CA, USA). Analysis conditions were according to [20 (link)]. The mobile phase consisted of a gradient of 1% phosphoric acid and acetonitrile (Table 1 ) with a flow rate of 0.5 mL per minute.
Samples were obtained by diluting the extract with methanol to the concentration of 4 mg/mL followed by filtration with a 0.45 μm filter (Millex, Merck KGaA, Darmstadt, Germany). Commercial standards were used in an attempt to identify the compounds present in HEMNL by comparison of retention times and ultraviolet spectra. The standards were caffeic acid, chlorogenic acid, ferulic acid, kaempferol, gallic acid, rosmarinic acid, ellagic acid, isoquercitrin, hesperetin, quercetin, resveratrol, vitexin, isovitexin, coumarin, and myricetin acquired from Sigma-Aldrich®, hyperoside acquired from Hwi Analytik Gmbh® (Rülzheim, Germany), and rutin from Chromadex® (Los Angeles, CA, USA).
Samples were obtained by diluting the extract with methanol to the concentration of 4 mg/mL followed by filtration with a 0.45 μm filter (Millex, Merck KGaA, Darmstadt, Germany). Commercial standards were used in an attempt to identify the compounds present in HEMNL by comparison of retention times and ultraviolet spectra. The standards were caffeic acid, chlorogenic acid, ferulic acid, kaempferol, gallic acid, rosmarinic acid, ellagic acid, isoquercitrin, hesperetin, quercetin, resveratrol, vitexin, isovitexin, coumarin, and myricetin acquired from Sigma-Aldrich®, hyperoside acquired from Hwi Analytik Gmbh® (Rülzheim, Germany), and rutin from Chromadex® (Los Angeles, CA, USA).
4
UHPLC-MS Analysis of Flavonoids
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UHPLC-MS grade solvents, formic acid, and LC-MS were from Merck (Santiago, Chile). Ultrapure water was obtained from a water system of purification (Milli-Q Merck Millipore, Chile). Jack Bean urease was purchased from Sigma-Aldrich (Santiago, Chile). Flavonol and anthocyanin standards (ellagic acid, rutin, quercetin, kaempferol, cyanidin, and cyanidin-3-O-glucoside, all standards with a high purity: 95% by HPLC) were acquired from ChromaDex (Santa Ana, CA, USA), Sigma-Aldrich (Saint Louis, Mo, USA), or Extrasynthèse (Genay, France).
5
Chamomile Phytochemical Characterization
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Nineteen samples of chamomile were obtained from different manufacturers in Poland and are listed in Table 1 with their characterizations. The samples were homogenized at 20°C in a water-cooled grinder Knifetec 1095 (Foss Tecator, Höganäs, Sweden) and stored in desiccator which was protected from light.
Standards of phenolic acids—gallic, caffeic, syringic, p-coumaric, ferulic, and of flavonoids—rutin, myricetin, quercetin and kaempferol were purchased from ChromaDex (California, USA). HPLC grade methanol and acetonitrile were purchased from POCh (Gliwice, Poland) and J.T. Baker (Phillipsbusg, USA), respectively. Analytical grade ethanol, methanol, acetic acid, ferric chloride hexahydrate, 6-hydroxy-2,5,7,8-tetramethychroman-2-carboxylic acid (Trolox), 2,4,6-tris (2-pyridyl)-1,3,5-triazine (TPTZ), and 1,1-diphenyl-2-picrylhydrazol radical (DPPH) were purchased from POCh (Gliwice, Poland). Trifluoroacetic acid (TFA) was obtained from Sigma Aldrich (St. Louis, MO, USA). Redistillated water was prepared by triple distillation of water in a Destamat Bi-18 system (Heraeus Quarzglas, Hanau, Germany).
Standards of phenolic acids—gallic, caffeic, syringic, p-coumaric, ferulic, and of flavonoids—rutin, myricetin, quercetin and kaempferol were purchased from ChromaDex (California, USA). HPLC grade methanol and acetonitrile were purchased from POCh (Gliwice, Poland) and J.T. Baker (Phillipsbusg, USA), respectively. Analytical grade ethanol, methanol, acetic acid, ferric chloride hexahydrate, 6-hydroxy-2,5,7,8-tetramethychroman-2-carboxylic acid (Trolox), 2,4,6-tris (2-pyridyl)-1,3,5-triazine (TPTZ), and 1,1-diphenyl-2-picrylhydrazol radical (DPPH) were purchased from POCh (Gliwice, Poland). Trifluoroacetic acid (TFA) was obtained from Sigma Aldrich (St. Louis, MO, USA). Redistillated water was prepared by triple distillation of water in a Destamat Bi-18 system (Heraeus Quarzglas, Hanau, Germany).
6
Extraction and Analysis of Artemisia Annua Compounds
7
Quantification of Phenolic Compounds
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Standards of phenolic acids such as gallic (GA), chlorogenic (CGA), caffeic (CA), p-coumaric (pCA), ferulic (FA) and of flavonoids such as rutin (RUT), myricetin (M), quercetin (Q), and kaempferol (K) were purchased from ChromaDex (CA, USA). HPLC-grade methanol was purchased from Avantor Performance Materials Poland S.A., and trifluoroacetic acid (TFA) was from Sigma-Aldrich (St. Louis, MO, USA).
8
Isolation and Characterization of Bioactive Compounds
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Caffeic acid, p-Coumaric acid, naringinen, chlorogenic acid, quercetin, isoquercetin, umbelliferone, harmine, luteolin, caffeine, 2,4,4′-trihydroxy chalcone, naringinin-7-glucoside, pseudoephedrine, glycyrrhizin, epicatechin, vanillic acid, gallic acid, β-glycyrrhetenic acid, xanthohumol and amentoflavone were purchased from Sigma Aldrich (St. Louis, MO, USA). Bisabolene, α-ionone, (Z)-3-hexenyl acetate, allyl anthranilate, isoeugenol, α-pinene, cuminaldehyde, thymol, caryophyllene oxide, carvone, and methyl disulphide were provided from Bedoukian Research (Danbury, CT, USA). Cynarin, emodin, rutin, and sennoside A were purchased from Chromadex (Wesel, Germany). Khellin was a gift from Prof. Dieter Treutter (Tech Univ. of Munich, Germany). Sarcophine was isolated from Sarcophyton ehrenbergi soft coral whereas vitexin and chrysin were isolated from Passiflora edulis leaf in Dr. Farag laboratory, Cairo University (Supplementary Fig. 6 ).
9
Analytical Characterization of Bioactive Compounds
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Analytical- and chromatographic-grade chemicals and solvents were used for this study: acetonitrile, methanol, glacial acetic acid, erythrodiol, maslinic, oleanolic acids from Sigma-Aldrich (GmbH, Karlsruhe, Germany); uvaol, betulin, corosolic acids, ursolic acid from Carl Roth (Karlsruhe, Germany); polyphenols (gallic acid, ellagic acid, neochlorogenic acid, chlorogenic acid, oenothein B, rutin, hyperoside, isoquercitrin, quercitrin, quercetin, guaijaverin, afzelin) were purchased from ChromaDex (Santa Ana, CA, USA), Sigma-Aldrich (Saint Louis, MO, USA), and Hwi Analytik (Rülzheim, Germany); all solvents used were of HPLC grade. Water was obtained using a Mili-Q purification system (Millipore, Burlington, MA, USA). Also, other reagents were used, such as sodium carbonate (Sigma-Aldrich, Scnelldorf, Germany), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), aluminium chloride, hexaethylenetetraamine, acetic acid obtained from Sigma-Aldrich (Buchs, Switzerland); potassium persulfate from Alfa Aesar (Karlsruhe, Germany); Trolox (98%) was received from Fluka Chemika (Buchs, Switzerland).
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