• Title/Summary/Keyword: HPLC analysis methods

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Isolation and Structural Identification of Antioxidant Substances from Ethyl Acetate Extract of Conyza canadensis (망초(Conyza canadensis) Ethyl Acetate 추출물의 항산화성 물질의 분리와 동정)

  • Hyun Sook Song
    • Journal of Naturopathy
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    • v.12 no.1
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    • pp.7-15
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    • 2023
  • Background: As a result of analyzing the components of wild Conyza canadensis, it contains physiologically active ingredients, so it is necessary to identify the compound. Purposes: It was to study the compound's molecular structure; a previous study showed that C. canadensis contains antioxidant substances. Methods: The ultrasonic pulverized lysate of C. canadensis stem and leaves was first extracted with 90% methanol and then five organic solvents. Next, the extracts was fractionated by HPLC, LC/MS chromatography, and NMR analyzers identified the molecular structure. Results: 100 g of dry C. canadensis was sonicated in 90% methanol and concentrated under reduced pressure to 11.96 g of a crude extract. Then, this crude was extracted with five types of solvents to obtain 123.8 mg of n-hexane, 448.2 mg of dichloromethane, 1047.7 mg of ethyl acetate (EA), 2563.8 mg of butanol, and 7.04 g of water. The EA extracts were fractionated by LC-MS and then re-fractionated to obtain F1 to F20. Next, the F15 was further fractionated to obtain nine fine fractions. Finally, the F17 fraction was re-fractionated to obtain ten fine fractions. As a result of LC-MS and NMR spectrometer analysis of the F15-7, the structure of this compound was confirmed as 3,5-dicaffeoylquinic acid. As a result of examining the structures of the F17-4 and F17-5 fractions, Quercetin-3-o-β-galactose was identified. In addition, the form of the F17-10 was confirmed to be 1,3,4-tri-caffeoylquinic acid. Conclusions: This study demonstrated that C. canadensis contained phenolic antioxidants, and its utilization may be expected.

In Vitro Antioxidant Activity and Anticonvulsant Properties on Zebrafish PTZ-Induced Seizure Model of a Tilia viridis Aqueous Extract

  • Elina Malen Saint Martin;Ayelen Morena Sosa;Carolina Soledad Martinez;Jimena Prieto;Carla Marrassini;Cecilia Beatriz Dobrecky;Maria Rosario Alonso;Claudia Anesini
    • Journal of Pharmacopuncture
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    • v.27 no.3
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    • pp.211-222
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    • 2024
  • Objectives: Tilia viridis (Bayer) Simonk. (Malvaceae) is widely distributed in Argentina and employed for its tranquilizing properties. Other species of the genus (Tilia europaea L., Tilia cordata Mill., Tilia platyphyllos Scop.) have been traditionally used for the treatment of epilepsy. Epilepsy affects approximately 65 million people worldwide and is characterized by an imbalance between excitatory and inhibitory processes in the brain, leading to unpredictable, unprovoked, recurrent seizures. Current pharmacological interventions often present mild to moderately severe side effects. Epilepsy has been associated with oxidative and nitrative stress as well as neuroinflammation. Herbal medicine therapies may offer new treatment options with multi-target antioxidant and anticonvulsant effects for patients whose seizures remain uncontrolled, potentially providing cost-effective solutions for individuals worldwide suffering from uncontrolled epilepsy.The aim of this study was to demonstrate the anticonvulsant activity of a standardized T. viridis aqueous extract (TE). Methods: Study of the constituents of TE, TE's antioxidant and anticonvulsant activities and toxicity, and analysis of the possible relation between the potential activities and the compounds present in the extract. In order to demonstrate TE's anticonvulsant activity a zebrafish model was used. The study also assessed TE's toxicity and antioxidant activity. To standardize the extract, total polyphenols and flavonoids were quantified and specific flavonoids were identified and quantified using HPLC-MS/MS and HPLC-UV. Results: TE exhibited anticonvulsant activity at low concentrations and demonstrated antioxidant effects by scavenging free radicals, exhibiting superoxide dismutase and peroxidase-like activities, as well as inhibiting lipoperoxidation. These actions can be attributed to the presence of polyphenols, particularly flavonoids. Conclusion: TE holds promise as a complementary herbal medicine in the treatment of epilepsy and may also offer benefits for other neuropathies associated with oxidative stress, such as Parkinson's disease and Alzheimer's disease.

Comparative Study on Anti-inflammatory Effects and Compound Contents of Multi-herbal Extracts HT008 produced by Different Manufacturing Methods (한약복합추출물 HT008의 제조방법에 따른 항염증 효능 및 성분함량 비교연구)

  • Lee, Donghun;Song, Jungbin;Song, Mikyung;Gaire, Bhakta Prasad;Park, Juyeon;Jin, Aihong;Choi, Ho-Young;Kim, Hocheol
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.71-76
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    • 2013
  • Objectives : The biological activities and compound contents of herbal medicine vary depending on manufacturing processes. In this study, we compared anti-inflammatory effects and compound contents of three kinds of multi-herbal extract HT008 produced by different manufacturing processes in order to determine chemical and biological equivalence. Methods : HT008 was produced by three different manufacturing methods: 1. Freeze dried extract of Eleutherococcus senticosus, Scutellaria baicalensis and Angelica sinensis (HT008 FD), 2. Spray dried extract of E. senticosus and S. baicalensis combined with reflux extract of A. sinensis (HT008 SD), 3. Spray dried extract of E. senticosus and S. baicalensis combined with supercritial fluid extract of A. sinensis (HT008 SF). Anti-inflammatory effects were evaluated using acetic acid induced pain model and ${\lambda}$-carageenan induced paw edema model. Compound contents were evaluated by HPLC quantitative analysis of standard compounds of HT008, eleutheroside E, baicalin, z-ligustilide. Results : HT008 FD, HT008 SD and HT008 SF significantly decreased acetic acid induced pain index and ${\lambda}$-carrageenan induced paw edema volume compared with that of control group. There was no significant difference in efficacy among the HT008 FD, HT008 SD and HT008 SF. Standard compound contents of HT008 FD, HT008 SD and HT008 SF were quantified within the range of Korean pharmacopoeia or other research. Conclusions : Three different manufacturing methods of multi-herbal extracts have been developed without noticeable difference in the efficacy or compound contents. The results might be used to establish manufacturing process and industrialization of herbal extracts.

Biotransformation of natural polyacetylene in red ginseng by Chaetomium globosum

  • Wang, Bang-Yan;Yang, Xue-Qiong;Hu, Ming;Shi, Li-Jiao;Yin, Hai-Yue;Wu, Ya-Mei;Yang, Ya-Bin;Zhou, Hao;Ding, Zhong-Tao
    • Journal of Ginseng Research
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    • v.44 no.6
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    • pp.770-774
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    • 2020
  • Background: Fermentation has been shown to improve the biological properties of plants and herbs. Specifically, fermentation causes decomposition and/or biotransformation of active metabolites into high-value products. Polyacetylenes are a class of polyketides with a pleiotropic profile of bioactivity. Methods: Column chromatography was used to isolate compounds, and extensive NMR experiments were used to determine their structures. The transformation of polyacetylene in red ginseng (RG) and the production of cazaldehyde B induced by the extract of RG were identified by TLC and HPLC analyses. Results: A new metabolite was isolated from RG fermented by Chaetomium globosum, and this new metabolite can be obtained by the biotransformation of polyacetylene in RG. Panaxytriol was found to exhibit the highest antifungal activity against C. globosum compared with other major ingredients in RG. The fungus C. globosum cultured in RG extract can metabolize panaxytriol to Metabolite A to survive, with no antifungal activity against itself. Metabolites A and B showed obvious inhibition against NO production, with ratios of 42.75 ± 1.60 and 63.95 ± 1.45% at 50 µM, respectively. A higher inhibitory rate on NO production was observed for Metabolite B than for a positive drug. Conclusion: Metabolite A is a rare example of natural polyacetylene biotransformation by microbial fermentation. This biotransformation only occurred in fermented RG. The extract of RG also stimulated the production of a new natural product, cazaldehyde B, from C. globosum. The lactone in Metabolite A can decrease the cytotoxicity, which was deemed to be the intrinsic activity of polyacetylene in ginseng.

LC-MS/MS analysis and anti-inflammatory effects of crude extract from Coptidis Rhizoma (황련 추출물의 LC-MS/MS 분석 및 항염증 효과)

  • Min-Jung, Kim;Ye-Jin, Yang;Kwang-Youn, Kim;Hun Hwan, Kim;Jae Dong, Son;Ju-Hye, Yang;Dong bin, Lee;Woo Hyun, Kim;Hu-Jang, Lee;Seon Been, Bak;Kwang-Il, Park
    • Herbal Formula Science
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    • v.31 no.1
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    • pp.1-10
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    • 2023
  • Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities. Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at -4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0-10 min at 10-15% B, 10-20 min at 20% B, 20-30 min at 25%, 30-40 min at 40%, 40-50 min at 70%, 50-60 min at 95%, and 60-70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 ㎍/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS. Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.

In vitro Antioxidative Activities and Phenolic Composition of Hot Water Extract from Different Parts of Cudrania tricuspidata

  • Jeong , Chang-Ho;Choi, Gwi-Nam;Kim, Ji-Hye;Kwak, Ji-Hyun;Heo, Ho-Jin;Shim, Ki-Hwan;Cho, Bok-Rai;Bae, Young-Il;Choi, Jine-Shang
    • Preventive Nutrition and Food Science
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    • v.14 no.4
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    • pp.283-289
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    • 2009
  • We evaluated total phenolics and antioxidative activities of water extracts from different parts of Cudrania tricuspidata (specifically, the leaves, stems, roots, and fruits). The antioxidative activities of these samples were determined using five methods, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical scavenging, reducing power, ferric reducing ability of plasma (FRAP), and $\beta$-carotene/linoleic acid system. The water extract of leaves exhibited the higher DPPH, ABTS radical scavenging activities, reducing power, and FRAP than water extract of stem, roots, and fruits. Inhibition values on linoleic oxidation of water extracts from leaves, stems, roots, and fruits were calculated as 45.98%, 33.03%, 39.73%, and 25.48% at 10 mg/mL, respectively. The water extract of C. tricuspidata leaves had the highest amount of toal phenolics (73.60$\pm$0.28 mg/g). High-performance liquid chromatography (HPLC) analysis showed that quercetin is the predominant phenolic compound in water extract of leaves. Thus, our study verified that the water extract of leaves has strong antioxidant activities which are correlated with its high level of phenolic compounds, particularly quercetin. This water extract of C. tricuspidata leaves can be used as an effective and safe source of antioxidants.

Re-evaluation of physicochemical and NMR data of triol ginsenosides Re, Rf, Rg2, and 20-gluco-Rf from Panax ginseng roots

  • Cho, Jin-Gyeong;In, Seo-Ji;Jung, Ye-Jin;Cha, Byeong-Ju;Lee, Dae-Young;Kim, Yong-Bum;Yeom, Myeonghun;Baek, Nam-In
    • Journal of Ginseng Research
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    • v.38 no.2
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    • pp.116-122
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    • 2014
  • Ginseng roots were extracted with aqueous methanol, and extracts were suspended in water and extracted successively with ethyl acetate and n-butanol. Column chromatography using the n-butanol fraction yielded four purified triol ginseng saponins: the ginsenosides Re, Rf, Rg2, and 20-gluco-Rf. The physicochemical, spectroscopic, and chromatographic characteristics of the ginsenosides were measured and compared with reports from the literature. For spectroscopic analysis, two-dimensional nuclear magnetic resonance (NMR) methods such as $^1H$-$^1H$ correlation spectroscopy, nuclear Overhauser effect spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond connectivity were employed to identify exact peak assignments. Some peak assignments for previously published $^1H$-and $^{13}C$-NMR spectra were found to be inaccurate. This study reports the complete NMR assignment of 20-gluco-Rf for the first time.

The Algicidal Activity of Arthrobacter sp. NH-3 and its Algicide against Alexandrium catenella and other Harmful Algal Bloom Species (Alexandrium catenella와 유해성 적조종에 대한 Arthrobacter sp. NH-3와 살조물질의 살조능)

  • Jeong, Seong-Yun;Jeoung, Nam Ho
    • Korean Journal of Environmental Agriculture
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    • v.34 no.2
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    • pp.139-148
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    • 2015
  • BACKGROUND: The aim of this study was to isolate and identify algicidal bacterium that tends to kill the toxic dinoflagellate Alexandrium catenella, and to determine the algicidal activity and algicidal range of algicide. METHODS AND RESULTS: Among of algicidal bacteria isolated in this study, NH-3 isolate was the strongest algicidal activity against A. catenella. NH-3 isolate was identified on the basis of biochemical characteristics and analysis of 16S rRNA gene sequences. The NH-3 isolate showed over 99% homology with Arthrobacter oxydans, and was designated as Arthrobacter sp. NH-3. The optimal culture conditions were $25^{\circ}C$, initial pH 7.0, and 2.0% (w/v) NaCl concentration. The algicidal activity of Arthrobacter sp. NH-3 was significantly increased to maximum value in the late of logarithmic phase. Arthrobacter sp. NH-3 showed algicidal activity through indirect attack, which excreted active substance into the culture filtrate. When 10% culture filtrate of NH-3 was applied to A. catenella, 100% of algal cells were destroyed within 30 h. In addition, the algicidal activities were increased in dose and time dependent manners. The pure algicide was isolated from the ethyl acetate extract of the culture filtrate of NH-3 by using silica gel column chromatography and high performance liquid chromatography (HPLC). We investigated the algicidal activity of this algicide on the growth of harmful algal bloom (HAB) species, including A. catenella. As a result, it showed algicidal activity against several HAB species at a concentration of $100{\mu}g/mL$ and had a relatively wide host range. CONCLUSION: Taken together, our results suggest that Arthrobacter sp. NH-3 and its algicide could be a candidate for controlling of toxic and harmful algal blooms.

A Study of MSDS Reliability Evaluation in Chemicals including Formaldehyde (포름알데히드 함유 화학제품의 MSDS 신뢰성 평가 연구)

  • Hong, Mun Ki;Song, Se Wook;Lee, Kwon Seob;Choi, Sung Bong;Lee, Jong Han
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.23 no.3
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    • pp.287-298
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    • 2013
  • Objectives: Workers who use chemicals are exposed to safety accidents and occupational diseases. Employers are required to provide workers with Material Safety Data Sheets (MSDSs) in order to prevent accidents and diseases related to chemicals. Thus, it is very important to offer reliable MSDSs. In this paper, we assessed the reliability of MSDSs for chemicals including formaldehyde. Methods: To evaluate MSDS reliability, we collected 14 MSDSs and bulk samples from the chemical industry. MSDS reliability was evaluated by the completeness of details. In order to evaluate the adequacy of the formaldehyde contents in a mixture, bulk samples were collected and analyzed by HPLC. The result of Globally Harmonized System (GHS) classification was confirmed by identifying physical chemical properties, toxicology information and ecological information. Results: The result of the evaluation of 14 MSDSs showed 76.29% average reliability on each item, especially 53.9% average appropriate rate on hazard risk classification. No chemicals failed to match between the content (%) in MSDSs and the result of analysis. Conclusions: To elevate MSDSs reliability, the certified education of MSDS drafters and reorganization of the MSDS circulation system is required.

Antidepressant Effect of the Subchronic Administration of the Methanolic Extract of Wild-ginseng and Cultivated-ginseng in Mice Tail Suspension Test (산삼과 인삼 메탄올 추출물 아만성 복용의 Mice Tail Suspension Test에서의 항우울 효과에 대한 비교연구)

  • Kwon, Sun-Oh;Choi, Soo-Min;Kim, Myung-Hwan;Lee, Bom-Bi;Park, Moo-Won;Lee, Hye-Jung;Park, Hi-Joon;Hahm, Dae-Hyun
    • Journal of Acupuncture Research
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    • v.26 no.4
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    • pp.99-106
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    • 2009
  • Objectives : The antidepressant effect of the subchronic administration of the methanolic extract of wild ginseng(WG) was investigated compared with that of cultivated ginseng(CG, panax ginseng) extract. Methods : To assess the antidepressant effect of the ginseng extracts, tail suspension test(TST) was executed in mice after daily administration of WG or CG extract for five consecutive days. Results : The WG extract at daily dose of 600mg/kg significantly reduced the total duration of immobility in the TST, whereas there was no significant reduction at daily dose of 300mg/kg WG and 600mg/kg CG. There were no individual differences between experimental groups in open field test (OFT) to evaluate psychostimulant effects of WG or CG extract. In the high performance liquid chromatography(HPLC) analysis of the extracts, it was found that WG included four times more ginsenoside Rg1 and Re, three times more Rf, and six times more Rb1 and Rc than CG. Conclusions : It is suggested that WG extract has stronger antidepressant effect than CG extract, which means it includes more antidepressant compounds than CG.

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