Identification and HPLC Quantification of a Phytoecdysone and Three Phenolic Glycosides in Lamium takesimense Nakai

The herbs of Lamium takesimense Nakai (Lamiaceae) is used to treat spasmodic and inflammatory disease. The four polar compounds, ecdysterone, isoacteoside, rutin and lamiuside C, were isolated and identified from the BuOH fraction of the L. takesimense MeOH extract. HPLC quantification was performed on a Capcell Pak C18 column (5 µm, 4.6 mm × 250 mm) with a gradient elution of H₂O and 0.05% acetic acid in MeOH. The HPLC method was validated in terms of linearity, sensitivity, stability, precision, and accuracy. The quantitative level in plant material was determined as the following order: lamiuside C (4, 3.75 mg/g dry weight) > ecdysterone (1, 1.93 mg/g) > isoacteoside (2, 1.32 mg/g) > rutin (3, 0.97 mg/g).

Isolation of Flavonoid Glycosides with Cholinesterase Inhibition Activity and Quantification from Stachys japonica

The three flavone glycosides, 4′-O-methylisoscutellarein 7-O-(6‴-O-acetyl)-β-D-allopyranosyl(1→2)-β-D-glucopyranoside (1), isoscutellarein 7-O-(6‴-O-acetyl)-β-D-allopyranosyl(1→2)-β-D-glucopyranoside (3), and isoscutellarein 7-O-β-D-allopyranosyl(1→2)-β-D-glucopyranoside (4) in addition to a flavonol glycoside, kaempferol 3-O-β-D-glucopyranoside (astragalin, 2), were isolated from Stachys japonica (Lamiaceae). In cholinesterase inhibition assay, compound 1 significantly inhibited aceylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities (IC₅₀s, 39.94 µg/ml for AChE and 86.98 µg/ml for BChE). The content of isolated compounds were evaluated in this plant extract by HPLC analysis. Our experimental results suggest that the flavonoid glycosides of S. japonica could prevent the memory impairment of Alzheimer's disease.

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

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 µg/mL and 23.08 µg/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 (28.15mg/g) > phlinoside (22.60mg/g) > leucosceptoside A (16.87 mg) > acteoside (7.05 mg/g).

Identification and quantification of flavonoids in Carica papaya leaf and peroxynitrite-scavenging activity

Objective To characterize the types, contents, and peroxynitrite-scavenging activities of flavonoids in the leaf of Carica papaya (C. papaya). Methods Chromatographic and spectroscopic techniques along with high performance liquid chromatography quantitative analysis and peroxynitrite-scavenging assay were performed to isolate and quantify flavonoid compounds in the flavonoid-rich fraction (BuOH fraction) derived from MeOH extract of C. papaya leaves and evaluate their peroxynitrite-scavenging activities. Results Seven flavonoids were isolated from the leaves of C. papaya, including quercetin 3-(2

Anti-acetylcholinesterase activity of the aglycones of phenolic glycosides isolated from Leonurus japonicus

Objective To find the genuine structure with anti-acetylcholinesterase (anti-AChE) from the phenolic glycosides abundant in Leonurus japonicus (Lamiaceae). The assay for anti-AChE activity is often used to lead anti-Alzheimer's drugs. Methods The five phenolic glycosides, tiliroside, leonurusoside C, 2‴-syringoylrutin, rutin, and lavanduliofolioside were isolated from L. japonicus. The activities of the glycosides were relatively low. Seven compounds including p-coumaric acid, caffeic acid, hydroxytyrosol, salidroside, syringic acid, kaempferol, and quercetin, which are produced by the hydrolysis of the five glycosides, were also assayed for anti-AChE activity. Results Of those seven compounds, the five compounds other than salidroside and syringic acid exhibited potent anti-AChE activities. In particular, the IC

Comparative Study on the Content and Cytotoxicity of Pseudolaric Acid B in the Five Plant Parts of Pseudolarix kaempferi

Pseudolaric acids of Pseudolarix kaempferi (Pinaceae) have been known as diterpenoids with potent anti-fungal-, anti-microbial, and cytotoxic activities. In the present study, the five MeOH extracts were prepared from the five plant part (root bark, stem bark, leaf, the inner part of root, and cone) to find the relation between the concentration of pseudolaric acids and cytotoxicity. Pseudolaric acids B and C were isolated from the root bark of P. kaempferi to use them as standard compounds. The five extracts were tested on cytotoxicity against six cancer cell lines, A549 (lung), HCT116 (colon), MDA-MB-231 (breast), SNU638 (stomach), and SK-hep-1 (liver) by SRB assay, but against K562 (leukemia) by SRB- or MTT assay. HPLC quantification were performed on a Shisheido Capcell PAK C18 column (5 µm, 4.6 mm × 250 mm) using 254 nm wavelength. The cytotoxicity (IC₅₀, 0.36 µg/ml on K562 cell lines) of the root bark extract was potent and the content (101.1 mg/g extract) of pseudolaric acid B was very high in the root bark. These results suggest that the MeOH extract obtained from the root bark could be developed as the anti-cancer agent with a high quantity of pseudolaric acid B.

Analysis of Flavonoid Composition of Korean Herbs in the Family of Compositae and their Utilization for Health

Compositional differences in flavonoids are varied in the big family of Compositae. By summarizing our previous analytical studies and other scientific evidences, new strategy will be possible to further analyze flavonoids and utilize them for human health. The HPLC analytical method has been established in terms of linearity, sensitivity, accuracy, and precision. Herbs of the family of Compositae have considerable amounts of peroxynitrite (ONOO-)-scavenging effects and their phenolic substances. These effects may contribute to the prevention of disease associated with excess production of ONOO-, depending on the high content of flavonoid substances.

HPLC analysis of Phenolic Substances and Anti-Alzheimer's Activity of Korean Quercus Species

This study aimed to establish the quantitative method to analyze the content of peroxynitrite-scavengers belonging to polyphenols in six Korean Quercus species (Quercus mongolica, Q. dentata, Q. acutissima, Q. alienta, Q. serrata, and Q. variabilis) by HPLC. The twelve peroxynitrite-scavengers, flavanols (catechins: (+)-catechin, (−)-epicatechin, and (−)-epigallocatechin), flavonols (kaempferol and quercetin), flavonol glycosides (astragalin, quercitrin, and isoquercitrin), flavonol acylated glycosides (astragalin 6″-gallate and isoquercitrin 6″-gallate), gallic acid and its dimer (ellagic acid) were analyzed by HPLC. Further, anti-Alzheimer's activity was assayed in a passive avoidance testusing mice by measuring the retention latency (sec), the concentration of acetylcholine (ACh), and acetylcholinesterase (AChE) activity. Simultaneous analysis of the extracts of the six Quercus leaves was achieved on a Capcell C18 column (5 µm, 250 mm × 4.6 mm i.d.) with a gradient elution of 0.05% HAc and 0.05% HAc in CH₃CN. In the extract of Q. mongolica leaves, the content of gallic acid (32.53 mg/g), (+)-catechin (28.78 mg/g), (−)-epicatehin (22.03 mg/g), astragalin 6″-gallate (20.94 mg/g), and isoquercitrin 6″-gallate (44.11 mg/g) and peroxynitrite-scavenging activity (IC₅₀, 0.831 µg/ml) were high. This extract delayed the retention latency and inhibited acetylcholinesterase activity in scopolamine-induced memory impairment of mice, suggesting that it has anti-Alzheimer's activity.

HPLC and GC-MS Analysis of Phenolic Substances in Acer tegmentosum

The stem barks, heartwoods, and leaves of Acer tegmentosum (Aceraceae) are widely used in Korea to treat hepatic or cerebral disorders mainly due to alcohol poisoning. This study was aimed to analyze phenolic substances in A. tegmentosum. Quantitative analysis of the three phenolic substances (salidroside, (+)-catechin and scopoletin) was performed by HPLC and the identification of volatile phenolic substances were done by GC-MS. The contents of the three compounds in the three MeOH extracts were higher in the stem bark (salidroside: 80.22 mg/g, (+)-catechin: 23.31 mg/g, and scopoletin: 9.45 mg/g) compared to the heartwoods and leaves. And GC-MS analysis of the stem bark extract demonstrated that p-tyrosol is a main substance of twenty-one compounds identified.

Quantitative Determination of Five Phenolic Peroxynitrite-scavengers in Nine Korean Native Compositae herbs

Peroxynitrite (ONOO-)-scavenging activities of nine Compositae herbs consisting of three Ixeris, two Youngia, two Cirsium and one of each Lactuca and Taraxacum species were evaluated. The contents of their ONOO- scavengers in the extracts were also determined on a HPLC using seven standard compounds, chlorogenic acid (CGA), chicoric acid (CA), luteolin 7-glucoside (luteolin-7-glc), luteolin 7-glucuronide (luteolin-7-glcU), luteolin, linarin and pectolinarin. Five of those compounds exhibited potent ONOO--scavenging activities: IC50, CA (0.76 microM), CGA (1.34 microM), luteolin (0.81 microM), luteolin-7-glc (0.86 microM) and luteolin-7-glcU (3.13 microM). Both CA and luteolin-7-glc were highly contained in I. dentata (19.71 mg/g and 13.58 mg/g, respectively), I. dentata var. albiflora (17.58 mg/g and 23.83 mg/g, respectively) and I. sonchifolia (65.71 mg/g and 6.99 mg/g, respectively). Among the nine herbs, those three Ixeris species had very low IC50 values over the range of 0.48 - 1.74 microg/mL, suggesting that they could be potential therapeutic vegetables, particularly for preventing diabetic complications or obesity, which can be caused by an excess production of ONOO-.

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

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 microm, 250 mm x 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.