Simultaneous analysis and peroxynitrite-scavenging activity of galloylated flavonoid glycosides and ellagic acid in Euphorbia supina

Nugroho, Agung,Rhim, Tae-Jin,Choi, Moo-Young,Choi, Jae Sue,Kim, Youn-Chul,Kim, Min-Sun,Park, Hee-Juhn 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.7

The herbs of Euphorbia supina (Euphorbiaceae) have been used to treat hemorrhage, chronic bronchitis, hepatitis, jaundice, diarrhea, gastritis, and hemorrhoids as a medicinal herb. This work is aimed to qualitatively and quantitatively analyze the polyphenols with peroxynitrite-scavenging activities. The eight compounds: gallic acid, methyl gallate, avicularin, astragalin, juglanin, isoquercitrin 6"-gallate, astragalin 6"-gallate, and ellagic acid, were isolated from E. supina and used for HPLC analysis and peroxynitrite ($ONOO^-$)-scavenging assay. Simultaneous analysis of the eight compounds was performed on MeOH extract and its fractions. The contents in MeOH extract and peroxynitrite-scavenging activities of the dimer of gallic acid, ellagic acid (15.64 mg/g; $IC_{50}$ $0.89{\mu}M$), and two galloylated flavonoid glycosides, astragalin 6"-gallate (13.72 mg/g; $IC_{50}$ $1.43{\mu}M$) and isoquercitrin 6"-gallate (16.99 mg/g; $IC_{50}$ $1.75{\mu}M$),were high, compared to other compounds. The legendary uses of E. supina could be attributed to the high content of polyphenols, particularly ellagic acid, isoquercitrin 6"-gallate, and astragalin 6"-gallate as active principles.

Phytochemical Studies of the Phenolic Substances in Aster glehni Extract and its Sedative and Anticonvulsant Activity

Agung Nugroho,박희준,Myung-Hoe Kim,최종원,최재수,정원태,이경태 대한약학회 2012 Archives of Pharmacal Research Vol.35 No.3

On high performance liquid chromatography, the caffeoylquinic acid (CQ) occupying the highest proportion of the water-ethanol (7:3) extract of Aster glehni (Compositae) leaves was 3-Op-coumaroylquinic acid (46.10 ± 4.22 mg/g of dried weight) among CQs tested. The IC50 of the water-ethanol (7:3) extract was 4.23 ± 0.24 μg/mL in the peroxynitrite (ONOO−)-scavenging assay. Phytochemical isolation from A. glehni extract yielded three kaempferol glycosides. The water-ethanol (7:3) extract and both p-coumaric acid and caffeic acid, phenylpropanoid moieties of CQs, had sedative effects in pentobarbital-induced sleeping time in mice and anticonvulsant effects in pentylenetetrazole-induced mice. Furthermore, the phenolic substance-rich W-E (7:3) extract of A. glehni could be used to treat anxiety or convulsion partly due to its peroxynitrite-scavenging mechanism.

Simultaneous analysis and peroxynitrite-scavenging activity of galloylated flavonoid glycosides and ellagic acid in Euphorbia supina

Agung Nugroho,임태진,최무영,최재수,김윤철,김민선,박희준 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.7

The herbs of Euphorbia supina (Euphorbiaceae)have been used to treat hemorrhage, chronic bronchitis, hepatitis,jaundice, diarrhea, gastritis, and hemorrhoids as amedicinal herb. This work is aimed to qualitatively andquantitatively analyze the polyphenols with peroxynitritescavengingactivities. The eight compounds: gallic acid,methyl gallate, avicularin, astragalin, juglanin, isoquercitrin600-gallate, astragalin 600-gallate, and ellagic acid,were isolatedfrom E. supina and used for HPLC analysis and peroxynitrite(ONOO-)-scavenging assay. Simultaneous analysis of theeight compounds was performed on MeOH extract and itsfractions. The contents in MeOH extract and peroxynitritescavengingactivities of the dimer of gallic acid, ellagic acid(15.64 mg/g; IC50 0.89 lM), and two galloylated flavonoidglycosides, astragalin 600-gallate (13.72 mg/g; IC50 1.43 lM)and isoquercitrin 600-gallate (16.99 mg/g; IC50 1.75 lM),werehigh, compared to other compounds. The legendary uses of E. supina could be attributed to the high content of polyphenols,particularly ellagic acid, isoquercitrin 600-gallate, and astragalin600-gallate as active principles.

In Vivo Sedative and Gastroprotective Activities of Salvia plebeia Extract and Its Composition of Polyphenols

Agung Nugroho,박희준,Myung-Hoe Kim,최종원,백남인 대한약학회 2012 Archives of Pharmacal Research Vol.35 No.8

Animal experiments were performed to develop Salvia plebeia (Labiatae) as a medicinal herb with sedative and gastroprotective activities; the former activity was measured using a pentobarbital-induced assay and the latter activity was measured in two gastric lesion-induced assays (HCl/EtOH-induced and indomethacin/bethanechol-induced assays) in mice. The MeOH extract and its EtOAc fraction were effective, although the former was less active than the latter. Rosmarinic acid (RA) isolated from S. plebeia was active in the same method at 10 and 20 mg/kg (p.o.). HPLC quantification demonstrated that RA comprised the largest proportion (28.5% of the MeOH extract, 33.0% of EtOAc extract; 4.46% of dry weight) of S. plebeia. The contents of five other compounds were much less than that of RA, although the contents of the three glycosides, 6-hydroxyluteolin 7-O-glucoside (0.28% of dry weight), cynaroside (0.35%) and nepitrin (0.43%) were higher than those of the two aglycones, quercetin (0.024%) and eupatilin (0.058%). The HPLC method was validated in terms of linearity, precision, accuracy and reproducibility. These results suggest that the main polyphenol, RA, plays a major role in the sedative and gastroprotective effects of S. plebeia.

Identification and quantification of the sedative and anticonvulsant flavone glycoside from Chrysanthemum boreale

Agung Nugroho,임상철,최종원,박희준 대한약학회 2013 Archives of Pharmacal Research Vol.36 No.1

The flowers or leaves of Chrysanthemumboreale (Compositae) have been traditionally used as herbtea to reduce anxiety, insomnia, and stress. Sedative andanticonvulsant activities were evaluated in mice usingpentobarbital-induced sleeping assay and pentylenetetrazole(PTZ)-induced convulsion assay. The flower extractexhibited more potent activities than the extracts of theleaves and stems, and chromatographic isolation yieldedthe five compounds acacetin, linarin, acacetin 7-O-b-Dglucopyranosyl-(1 ? 2)[a-L-rhamnopyranosyl-(1 ? 6)]-b-D-glucopyranoside, chlorogenic acid, and 3,5-di-O-caffeoylquinic acid. These compounds were simultaneouslyanalyzed by HPLC, and the method was validated. Thecontents of linarin, which were shown to be most abundant inC. boreale, were observed in the order of leaf (11.93 mg/g)[flower (8.50 mg/g)[stem (5.60 mg/g). Linarin and itsaglycone, acacetin, exhibited sedative and anticonvulsantactivities in the present in vivo assays. It can be consideredthat linarin is one of the active compounds effective againstanxiety, insomnia, and stress, with acacetin as its activemoiety.

Isolation and Quantitative Analysis of Peroxynitrite Scavengers from Artemisia princeps var. orientalis

Agung Nugroho,Kang Ro Lee,Md. Badrul Alam,최재수,박희준 대한약학회 2010 Archives of Pharmacal Research Vol.33 No.5

Young and mature Artemisia princeps var. orientalis (APO, Compositae) are used as a health food and a medicinal plant, respectively, in Korea. Here, we identified the in vitro potent peroxynitrite (ONOO-)-scavenging effect (IC50, 0.26 μg/mL) of the components from the EtOAc fraction. Octadecylsilane column chromatography on the EtOAc fraction yielded two caffeoylquinic acid compounds, 3,5-di-O-caffeoyl-muco-quinic acid (1) and methyl 4,5-di-O-caffeoylquinate (2) by NMR spectroscopic data, which have not been reported before from APO. The IC50 values of compounds 1 and 2 were 0.18 ± 0.01 μg/mL and 0.12 ± 0.00 μg/mL, respectively, lower than that of the positive control (L-penicillamine). HPLC data indicated that young APO (1: 30.3 mg/g dried weight, 2: 27.7 mg/g) contained considerably higher quantities of the two caffeoylquinic acids than mature APO (1: 1.77 mg/g dried weight, 2: 4.10 mg/g).

Analysis of Essential oil, Quantification of Six Glycosides, and Nitric Oxide Synthase Inhibition Activity in Caryopteris incana

Agung Nugroho,이상국,김동화,최재수,박경식,송병민,박희준 한국생약학회 2018 Natural Product Sciences Vol.24 No.3

Caryopteris incana (Verbenaceae) has been used to treat cough, arthritis, and eczema in Oriental medicine. The two fractions (CHCl3- and BuOH fractions) and the essential oil of the plant material were subjected to the inducible nitric oxide synthase (iNOS) assay. The IC50 of the CHCl3 fraction and the essential oil on LPS-induced macrophage RAW 264.7 cells were 16.4 mg/mL and 23.08 mg/mL, respectively. On gas chromatography (GC)-mass spectroscopy (MS) analysis, twenty-five components representing 85.5% amount of total essential oil were identified. On the chromatogram, three main substances, trans-pinocarveol, cis-citral, and pinocarvone, occupied 18.8%, 13.5% and 18.37% of total peak area. Furthermore, by HPLC-UV analysis, six compounds including one iridoid (8-O-acetylharpagide)- and five phenylethanoid glycosides (caryopteroside, acteoside, phlinoside A, 6-O-caffeoylphlinoside, and leucosceptoside A) isolated from the BuOH fraction were quantified. The content of six compounds were shown as the following order: caryopteroside (162.35 mg/g) > 8-O-acetylharpagide (93.28 mg/g) > 6-O-caffeoylphlinoside (28.15 mg/g) > phlinoside (22.60 mg/g) > leucosceptoside A (16.87 mg) > acteoside (7.05 mg/g).

Qualitative and Quantitative Determination of the Caffeoylquinic Acids on the Korean Mountainous Vegetables Used for Chwinamul and Their Peroxynitrite-Scavenging Effect

Agung Nugroho,김기현,Kang Ro Lee,Md. Badrul Alam,최재수,Won-Bae Kim,박희준 대한약학회 2009 Archives of Pharmacal Research Vol.32 No.10

Mountainous vegetables called chwinamul are used in Korea to promote health. Chwinamul was obtained from several plants belonging to the Compositae - e.g., Kalimeris yomena, Aster scaber, Solidago virga var. gigantea, Solidago viragaurea var. asiatica, Saussurea grandifolia,Ainsliaea acerifolia - were used for our experiments. Analytical methods for simultaneous determination of the caffeoylquinic acids (3,4-di-O-caffeoylquinic acid, 3,5-di-O-dicaffeoyl-epiquinic acid, 3,5-di-O-caffeoylquinic acid, 4,5-di-O-caffeoylquinic acid, 5-O-caffeoylquinic acid,3-O-caffeoylquinic acid, 3-O-p-coumaroyl-caffeoylquinic acids) were established for chwinamul. The kinds of constituents were identified from HPLC chromatograms and it was possible to calculate the percentage (w/w) of seven of these compounds in the dried plants and in the extracts. The proportion of caffeoylquinic acids in the extracts ranged from 20.25 to 38.35%. Since it is known that peroxynitrite (ONOO−)-scavenging is beneficial for amelioration of obesity, diabetes mellitus, atherosclerosis and even Alzheimer’s disease, assays for peroxynitritescavenging activity were performed on the seven chwinamul plants. Of the tested extracts, theMeOH extract of A. acerifolia had the most potent effect (IC50 1.49 ± 0.68 μg/mL). These results suggest that chwinamul vegetables can be used for treatment or prevention of peroxynitrite-related diseases.

The Inhibitory Effect of the Caffeoylquinic acid-Rich Extract of Ligularia stenocephala Leaves on Obesity in the High Fat Diet-Induced Rat

Agung Nugroho,Moch Saiful Bachri,최종원,최재수,Won-Bae Kim,Byung-Il Lee,김종대,박희준 한국생약학회 2010 Natural Product Sciences Vol.16 No.2

High performance liquid chromatography (HPLC) analysis on the MeOH extract of Ligularia stenocephala leaves identified six caffeoylquinic acids, viz. 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoyl-muco-quinic acid, 3,5-di-O-caffeoylquinic acid, 4,5-di-O-caffeoylquinic acid, 5-O-caffeoylquinic acid, and 3-O-caffeoylquinic acid using standard compounds, and determined the quantity of each extract. Percentage of total caffeoylquinic acids of the MeOH extract and its BuOH fraction were calculated as 67.83 ? 3.79% and 94.52 ? 1.84%, respectively. Since the caffeoylquinic acid-rich MeOH extract exhibited a potent peroxynitrite-scavenging effect in vitro (IC50 = 0.87 ? 0.33 mg/ml (mean ? SEM)), the experiment was designed to identify whether or not that extract has an anti-obesity effect on rat obesity induced by high fat diet. Oral administration of the MeOH extract and its BuOH fraction abundant in caffeoylquinic acid decreased the rat body weight to the level of untreated group and decreased abdominal fat pad weight. The atherogenic index and thiobarbituric acid-reactive substance (TBARS) values were restored by treatment, indicating that the caffeoylquinic acid-rich extract probably inhibited hyperlipidemia and oxidative stress caused by high fat diet. These results suggest that L. stenocephala in vegetable form or its caffeoylquinic acid-rich fraction (BuOH fraction) as an agent can be used for treatment or prevention of obesity.

Phytochemical Analysis of the Phenolic Fat-Suppressing Substances in the Leaves of Lactuca raddeana in 3T3-L1 Adipocytes

Agung Nugroho,최재수,안효진,박희준 한국생약학회 2015 Natural Product Sciences Vol.21 No.1

Lactuca raddeana (Compositae) is used to treat obesity and complications due to diabetes. The five phenolic compounds including chlorogenic acid, chicoric acid, luteolin 7-O-glucoside, luteolin 7-O-glucuronide, luteolin were qualitatively identified by LC-ESI-MS analysis. The contents were quantitatively determined by HPLC, under the condition of a Capcell Pak C18 column (5 mm, 250 mm ? 4.6 mm i.d.) and a gradient elution of 0.05% trifluoroacetic acid (TFA) and 0.05% TFA in MeOH-H2O (60 : 40). The contents of chicoric acid (100.99 mg/g extract) and luteolin 7-O-glucoside (101. 69 mg/g extract) were high, while those of other three phenolic substances were very low. The 3T3-L1 adipocyte cells treated with chicoric acid and luteolin 7-O-glucuronide significantly suppressed the accumulation of fat, suggesting they are effective against obesity. Since high level of peroxynitrite (ONOO) causes cardiovascular disease in obese patients, its scavenging activity was also studied.