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2 protocols using corosolic acid

1

Comprehensive Analytical Standards Protocol

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Methanol (MeOH) and hexane (HEX), both AnalaR
Normapur, dichloromethane (DCM) and ethyl acetate (EtOAc), both GPR
Rectapur, and acetonitrile (ACN) HiPerSolv Chromanorm for HPLC–CAD–MS
analysis were obtained from VWR (Vienna, Austria). Formic acid (purity
>98.0%) and conc. ammonia (24%) were purchased from Gatt-Koller
(Absam,
Austria). Water, for extraction and phytochemical analysis, was deionized
and distilled. Betulinic acid (purity 95%), corosolic acid (purity
90%), maslinic acid (purity 89%), oleanolic acid (purity 94%) and
ursolic acid (purity 94%), cyanidin-3-O-sambubioside
(purity 99%), cyanidin-3-O-glucoside (purity 88%),
delphinidin-3-O-glucoside (purity 87%), malvidin-3-O-glucoside (purity 99%), peonidin-3-O-glucoside
(purity 94%), and petunidin-3-O-glucoside (purity
99%) were provided from Phytolab (Vestenbergsgreuth, Germany), hyperoside
(purity 99%), and rutin (purity 99%) from Extrasynthèse (Lyon,
France), and palmitic acid (purity 99%) and linoleic acid (purity
99%) from Sigma-Aldrich (St. Louis, MO).
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2

Quantification of Triterpene Acids

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Maslinic acid, corosolic acid, oleanolic acid, and ursolic acid were obtained from Phytolab (Vestenbergsgreuth, Germany), lithocholic acid (LCA), chenodeoxycholic acid (CDCA), oleic acid, and palmitic acid from Sigma Aldrich (St. Louis, MO, United States), stearic acid and 4-methoxycinnamic acid from Fluka (Buchs, Switzerland), chlorogenic acid from Carl Roth (Karlsruhe, Germany), and linoleic acid from Acros (Geel, Belgium). Eugenol was isolated from clove and the identity and purity tested by ATR-IR spectroscopy. All reference and standard compounds were obtained in a purity of ≥90%. They were dissolved to 1.0 mg/mL in MeOH. The triterpene acids (TTAs) used for the quantification were mixed and diluted to give a standard solution containing 100 μg/mL of each corosolic acid, Maslinic acid, oleanolic acid, and ursolic acid followed by a serial dilution down to 2 μg/mL.
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