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).
Caffeic acid
Manufactured by ChromaDex
Sourced in Germany, United States
Caffeic acid is a natural compound that serves as a key laboratory analytical standard. It is used to quantify and identify the presence of this compound in various samples during chemical analysis.
Automatically generated - may contain errors
Lab products found in correlation
Chlorogenic acid, by ChromaDex (7 mentions)
Ferulic acid, by ChromaDex (4 mentions)
Gallic acid, by ChromaDex (3 mentions)
P coumaric acid, by ChromaDex (3 mentions)
Methanol, by Carl Roth (2 mentions)
Acetonitrile, by Carl Roth (2 mentions)
Cinnamic acid, by ChromaDex (2 mentions)
Kaempferol, by ChromaDex (2 mentions)
Milli q plus water purification system, by Merck Group (2 mentions)
Rutin, by ChromaDex (1 mentions)
Apigenin, by ChromaDex (1 mentions)
Ellagic acid, by ChromaDex (1 mentions)
Astragalin, by ChromaDex (1 mentions)
Mangiferin, by ChromaDex (1 mentions)
Isoorientin, by ChromaDex (1 mentions)
Biochanin a, by ChromaDex (1 mentions)
Sinensetin, by ChromaDex (1 mentions)
Rosmarinic acid, by ChromaDex (1 mentions)
Eupatorin, by ChromaDex (1 mentions)
Lc ms grade acetonitrile, by Merck Group (1 mentions)
9 protocols using caffeic acid
1
Analytical Standards for Phytochemicals
2
HPLC Analysis of Bioactive Compounds
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In order to determine the bioactive compound present in the plant extract, a standardized HPLC method was utilized, which has been described in the USP 30-NF25 Pharmacopoeia [49 ]. The HPLC system used consisted of an Agilent 1200 chromatogram equipped with a quaternary pump, DAD, thermostat, degas system, and autosampler (Agilent Tehnologies, Boblingen. Germany). A C18 Zorbax XDB column (250 mm × 4.6 mm; 5 µm) was employed, and the eluents were composed of 0.1% phosphoric acid (A) and acetonitrile (B), with a linear gradient of 10% B for 13 min, 22% B for 1 min, 40% B for 3 min, and 10% B for 1 min. The flow rate was set to 1.5 mL/min and the column temperature was maintained at 35 °C. The DAD system was set to detect wavelengths of 310 nm, 335 nm, and 360 nm simultaneously. The standards used in the study were purchased from Chromadex (Wesel, Germany) and included E-resveratrol, Z-resveratrol, caffeic acid, chlorogenic acid, cinnamic acid, ellagic acid, vanillin, gallic acid, ferulic acid, astragalin, isorhamnetin, kaempferol, scutellarin, rutoside, and quercetin. All assays were run in triplicate, and the results were expressed as mean ± SD.
3
HPLC Analysis of Phenolic Compounds
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The acetonitrile and methanol were of HPLC grade and were purchased from Roth GmbH (Karlsruhe, Germany). The reference compounds (chlorogenic acid, caffeic acid, ferulic acid, t-cinnamic acid, mangiferin, kaempferol, tectoridin, iristectorigenin B, nigricin, irigenin, isoorientin, genistein-7-glucoside, and biochanin A) were purchased from ChromaDex (Santa Ana, CA, USA), Sigma-Aldrich (Saint Louis, MO, USA), HWI ANALYTIK GmbH, and Roth GmbH (Karlsruhe, Germany). All other chemicals were of analytical grade.
4
Analytical UHPLC of Botanical Compounds
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Formic acid, methanol and LCMS grade acetonitrile were purchased from MERCK (Malaysia). HPLC grade water was prepared from distilled water using a Milli-Q-system (Millipore, MA,USA) and was used during analytical UHPLC analysis. Standards used were rosmarinic acid (Chromadex, CA, USA), caffeic acid, chlorogenic acid, eupatorin and sinensetin (Extrasynthase, Genay, France). All of other solvents and chemicals used in this study were analytical grade. Stock and working standards were prepared by dissolving these analytes in 100% methanol. The standard solutions stored at 4°C were stable for at least 3 months.
5
Cytochrome P450 Inhibition Kinetics Assay
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Inhibition and kinetic reactions were carried out in Black costar 96-well plates bought from Thermo Fischer Scientific (Pittsburgh, PA, USA). α-naphthoflavone, furafylline, sulphaphenazole, and miconazole were purchased from Sigma-Aldrich (St. Louis, MO, USA). Vivid® Blue screening kits and vivid® substrates, 7-benzyl-oxymethyloxy-3-cyanocoumarin, (BOMCC-cat. no. P2975) and 7-ethoxy-methloxy-3-cyanocoumarin (EOMCC-cat. no. P3024) were obtained from Life Technologies, (Grand Island, NY, USA). CYP2C19 (cat. no. P2864), CYP2C9 (cat. no. P2861), and CYP1A2 (cat. no. P2863) blue screening kit included baculosome (respective isozymes and NADPH-P450 reductase); a regeneration system (glucose-6-phosphate, glucose-6-phosphate dehydrogenase) and NADP+ were used for the study. Standards for epicatechin, catechin, chlorogenic acid, caffeic acid, and p-coumaric acid for identification and quantification of constituents of herbal extracts were purchased from Chromadex (Wesel, Germany). Ultra-Pure double distilled and deionized water was obtained from Milli-Q plus water purification system (Millipore, Bedford, MA, USA). All reagents were of analytical grade.
6
Quantification of Phenolic Acids in Plants
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Methanol, acetonitrile, and acetic acid, all HPLC grade were purchased from J.T. Baker (Phillispsburg, NJ). Chlorogenic acid and caffeic acid standards were purchased from Chromadex (Irvine, CA). Neo‐chlorogenic and crypto‐Chlorogenic acids were purchased from PhytoLab (Vestenbergsgreuth, Germany). All standards were prepared as stock solutions at 1.00 mg/mL in methanol and stored at −20°C until use. HPLC‐grade water was obtained using a Millipore system (Billerica, MA).
7
Analytical Characterization of Bioactive Compounds
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Formic acid, acetonitrile, methanol, and anhydrous sodium sulphate were purchased from J.T. Baker (Baker Mallinckrodt, Mexico), while diethyl ether, methyl tert-butyl ether (MTBE), β-carotene (97%), gallic acid and rutin (94%) were procured from Sigma-Aldrich (St. Louis, MO). The HPTLC plates Si 60 F254 (10 × 20 cm) were purchased from E. Merck (Darmstadt, Germany). The myricetin (98%), trans-ferulic acid (99%), caffeic acid (98%), and kaempferol (98%) were purchased from Chromadex (Life Technology, Bangalore, India). Solvents used for chromatography were of HPLC grade, and distilled water was obtained by a Milli-Q plus water purification system (Millipore Corp., Bedford, MA). Edoxin (cyclophosphamide injection) was procured from Apollo Pharmacy.
8
Assay for Cytochrome P450 Activity
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Miconazole nitrate was purchased from Sigma-Aldrich (St. Louis, MO, USA). Black costar 96 well plates were obtained from Thermo Fischer Scientific (Pittsburgh, PA, USA). Vivid® CYP450 Blue Screening Kit and Vivid® Substrate, 7-benzyl-oxymethyloxy-3-cyanocoumarin, (BOMCC) were purchased from Life Technologies, (Grand Island, NY, USA). Standard for epicatechin, catechin, chlorogenic acid, caffeic acid and p-coumaric acid for identification of constituents of herbal extracts were purchased from Chromadex (Wesel, Germany). Purified water (double-distilled and deionized) was obtained from Millipore (Bedford, MA, USA).
9
Quantitative Analysis of Phytochemicals
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Chemicals and reagents: Syringin, eleutheroside E, isofraxidin, hyperoside, rutin, kaempferol, oleanolic acid, and L-phenylalanine were purchased from the Chinese National Institute of Control of Pharmaceutical and Biological Products (Beijing, China). Syringic acid, vanillic acid, p-hydroxybenzoic, protocatechuic acid, cinnamic acid, gallic acid, ferulic acid, chlorogenic acid, caffeic acid, p-coumaric acid, luteolin, genistein, and quercetin were purchased from ChromaDex Inc. (Santa Ana, CA, USA). The water used for UPLC-MS/MS was prepared by a Milli-Q (Millipore, Bedford, MA, USA). Acetonitrile (J&K Scientific Ltd., Beijing, China) was HPLC grade. All other chemicals used in the research were of analytical grade.
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