Amarogentin

Manufactured by ChromaDex

Sourced in United States

Amarogentin is a chemical compound that serves as a key analytical reference standard. It is a colorless crystalline solid used to identify and quantify the presence of specific compounds in various analytical applications.

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5 protocols using amarogentin

Bitter Compound Extraction and Sourcing

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Absinthin and Parthenolide were available from previous studies (Brockhoff et al., 2007 (link)). Other reagents were purchased as follows: Amarogentin from ChromaDex; Limonin from Apin Chemical; Quassin from CPS Chemie and all other bitter tastants from Sigma-Aldrich.

Kumar P., Redel U., Lang T., Korsching S.I, & Behrens M. (2023). Bitter taste receptors of the zebra finch (Taeniopygia guttata). Frontiers in Physiology, 14, 1233711.

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Pharmacological Evaluation of Bitter Compounds

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Six structurally diverse bitter substances, found at low concentrations in various beverages, and known to exhibit pharmacological properties at high concentrations, were used to probe phenotypic and molecular responses (S1 Fig). These included absinthin (a dimeric sesquiterpene lactone), Amarogentin (a secoiridoid glycoside), cascarillin (a diterpene lactone), grosheimin (a sesquiterpene lactone), quassin (a triterpene lactone), and quinine (a quinoline alkaloid). Absinthin, cascarillin, and grosheimin were isolated from crude vegetable material as detailed in previous studies [31 (link), 56 (link)]. Amarogentin and quassin were purchased from Chromadex Inc. (Irvine, CA, USA), quinine hydrochloride from Sigma-Aldrich Co. (St. Louis, MO, USA). Salicin, sodium chloride, citric acid, and sucrose (Sigma-Aldrich Co.) were used as reference tastants for assessing bitter, salty, sour, and sweet perception, respectively.

Roudnitzky N., Behrens M., Engel A., Kohl S., Thalmann S., Hübner S., Lossow K., Wooding S.P, & Meyerhof W. (2015). Receptor Polymorphism and Genomic Structure Interact to Shape Bitter Taste Perception. PLoS Genetics, 11(9), e1005530.

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Amarogentin-Induced Signaling Pathways

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Amarogentin (88.9%) was purchased from ChromaDex (Irvine, CA). Arachidonic acid (AA), collagen (type I), 9,11-dideoxy-11α,9α-epoxymethanoprostaglandin (U46619), luciferin-luciferase, thrombin, SQ22536, phorbol-12,13-dibutyrate (PDBu), and 1H-[1,2,4]qxadiazolo[4,3-a]quinoxalin-1-one (ODQ) were purchased from Sigma (St. Louis, MO). The anti-phospho-c-Jun N-terminal kinase (JNK) (Thr¹⁸³/Tyr¹⁸⁵) and anti-phospho-p38 mitogen-activated protein kinase (MAPK) monoclonal antibodies (mAbs), and the anti-phospho-p44/p42 extracellular signal-regulated kinase (ERK) (Thr²⁰²/Tyr²⁰⁴), anti-phospholipase Cγ2 (PLCγ2), and anti-phospho (Tyr⁷⁵⁹) PLCγ2 polyclonal antibodies (pAbs) were purchased from Cell Signaling (Beverly, MA). The anti-phospho-p38 MAPK Ser¹⁸² mAb was purchased from Santa Cruz (Santa Cruz, CA). The anti-α-tubulin mAb was purchased from NeoMarkers (Fremont, CA). The anti-phospho-Akt (Ser⁴⁷³) and anti-Akt mAbs were purchased from Biovision (Mountain View, CA). The horseradish peroxidase- (HRP-) conjugated donkey anti-rabbit immunoglobulin G (IgG), the Hybond-P polyvinylidene difluoride (PVDF) membrane, the sheep anti-mouse IgG, and the enhanced chemiluminescence western blotting detection reagent were purchased from Amersham (Buckinghamshire, UK). The Amarogentin was dissolved in DMSO and stored at 4°C.

Yen T.L., Lu W.J., Lien L.M., Thomas P.A., Lee T.Y., Chiu H.C., Sheu J.R, & Lin K.H. (2014). Amarogentin, a Secoiridoid Glycoside, Abrogates Platelet Activation through PLCγ2-PKC and MAPK Pathways. BioMed Research International, 2014, 728019.

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Phytochemical Profiling and Antioxidant Analysis

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2,2-diphenyl-1-picrylhydrazyl (DPPH), sodium carbonate, ascorbic acid and folin–ciocalteu phenol reagent was purchased from Merck Ltd, India; gallic acid from Moly Chem, Mumbai, quercetin from Sigma-Aldrich and Aluminium TLC plates of 250 microns were purchased from Whattmann, Germany. The marker compound, Amarogentin was obtained from Chromadex USA; Mangiferin and Swertiamarin were obtained from ZeLang Pharma, Nanjing, China. All the reagents and chemicals including the solvents used in this experiment were of analytical grade.

Khanal S., Shakya N., Thapa K, & Pant D.R. (2015). Phytochemical investigation of crude methanol extracts of different species of Swertia from Nepal. BMC Research Notes, 8, 821.

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Immunohistochemical Analysis of CerS3 Expression

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The polyclonal rabbit-anti-human CerS3 antibody was from Antikörper-online (Aachen, Germany; 1:50). The secondary antibodies were the swine anti-rabbit-FITC antibody (Dako, Hamburg, Germany; 1:30) and the donkey anti-rabbit-Alexa-Fluor 555 antibody (Thermo Fisher Scientific, Darmstadt, Germany). DAPI was from Sigma-Aldrich GmbH (Taufkirchen, Germany). The following specific inhibitors were used: PPARγ antagonist (GW9962, Enzo, Lörrach, Germany), PI3K inhibitor (LY294002; Sigma-Aldrich GmbH), ERK inhibitor (PD98059, New England Biolabs GmbH, Frankfurt, Germany), p38 MAPK inhibitor (SB203580, Sigma-Aldrich GmbH) and NFκB inhibitor (GIV 3727, Merck Millipore, Darmstadt, Germany). Methanolic sodium hydroxide solution, hexane standards, loganic acid, and loganin were from Carl Roth GmbH (Karlsruhe, Germany). Boron trifluoride–methanol complex was from Merck KGaA (Darmstadt, Germany), and standard methyl heptadecanoate was from Sigma-Aldrich GmbH. Orto-phosphoric acid, 85%, Ph. Eur. p.a. grade was supplied by VWR. Amarogentin and gentiopicroside were purchased from Chromadex Inc. (Santa Ana, CA, USA).

Wölfle U., Haarhaus B., Seiwerth J., Cawelius A., Schwabe K., Quirin K.W, & Schempp C.M. (2017). The Herbal Bitter Drug Gentiana lutea Modulates Lipid Synthesis in Human Keratinocytes In Vitro and In Vivo. International Journal of Molecular Sciences, 18(8), 1814.

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