• Title/Summary/Keyword: HPLC Chromatography

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Effects of Caffeic Acid Methyl Ester from Mentha arvensis Linne var. piperascens on Function of E6 Oncoprotein of HPV 16 (박하에서 분리한 Caffeic Acid Methyl Ester가 자궁경부암 바이러스 발암단백질 E6의 기능에 미치는 영향)

  • Baek, Tae-Woong;Lee , Kyung-Ae;Ahn , Min-Jung;Joo, Hae-Hyung;Oh , Hyun-Cheol;Ahn , Jong-Seok;Cho , Yong-Kweon;Myung , Pyung-Keun;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.328-334
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    • 2004
  • Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

Production of Flavonoid Compounds and Anti-inflammatory Property of Fermented Licorice Extract with the Basidiomycete Grifola frondosa HB0071 (잎새버섯 균사체를 이용한 감초추출발효물의 플라보노이드 생성과 항염 활성 연구)

  • Bae, Jun Tae;Song, Min Hyeon;Kim, Jin Hwa;Lee, Geun Soo;Pyo, Hyeong Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.4
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    • pp.327-338
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    • 2012
  • Liquiritigenin and isoliquiritigenin are the major flavonoids present in licorice. These flavonoid compounds were prepared by submerged culture of Grifola frondosa (G. frondosa) HB0071 mycelia producing ${\beta}$-glucosidase in the aqueous extract of licorice. The contents of liquiritigenin and isoliquiritigenin were increased during the fermentation. This fungus produced a high ${\beta}$-glucosidase (activity of 91.5 mU/mL), thereby achieving high amounts of liquiritigenin and isoliquiritigenin ($568.5{\mu}g/mL$ and $89.6{\mu}g/mL$), respectively at 96 h. A reversed- phase high-performance liquid chromatography method was established for simultaneous determination of liquiritigenin and isoliquiritigenin in fermented licorice extract (FLEx). The anti-inflammatory activities were investigated by licorice extract (LEx) before and after fermentation with G. frondosa HB0071. The treatment of UVB-irradiated HaCaT keratinocytes with FLEx resulted in a dose-dependent decrease in the expression level of cyclooxygenase-2 (COX-2) mRNA. Furthermore, FLEx dose-dependently decreased mRNA of the pro-inflammatory cytokines of IL-$1{\beta}$ and IL-6 in UVB-irradiated HaCaT cells. These results suggest that FLEx may mitigate the effects of skin inflammation by reducing UVB-induced adverse skin reactions.

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.

Toxicokinetics of rifapentine in beagle dogs (Beagle dog에 있어서 rifapentine의 독성동태연구)

  • Shin, Ho-chul;Lee, Hye-suk;Cha, Shin-woo;Han, Sang-seop;Roh, Jung-ku;Kim, Jin-suk;Lee, Won-chang
    • Korean Journal of Veterinary Research
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    • v.35 no.4
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    • pp.815-822
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    • 1995
  • The toxicokinetics of rifapentine was studied after an oral administration to beagle dogs. High-performance liquid chromatography(HPLC) using column-switching technique was performed to determine the serum concentrations of rifapentine. The pharmacokinetic profiles of rifapentine were analysed using one-compartment open model. Following a single oral administration of 10mg/kg, pharmacokinetic parameters were determined as follows: maximum serum concentration($C_{max}$), $28.90{\mu}g/ml$; maximum concentration time($T_{max}$), 3.7hr; elimination half-life($t_{1/2}$, 4.7hr; area under the curve(AUC), $339.0{\mu}g{\cdot}hr/ml$; volume of disiribution/bioavailability (Vd/F), 0.21 l/kg; lag time, 24min; absorption rate constant($k_a$), $0.445hr^{-1}$; elimination rate constant($k_{el}$), $0.148hr^{-1}$. After 6 month multiple oral doses of 10mg/kg/day, parameters were as follows: $C_{max}$, $34.40{\mu}g/ml$; $T_{max}$, 2.6hr; $t_{1/2}$, 6.7hr; AUC, $391.3{\mu}g{\cdot}hr/ml$; Vd/F, 0.291/kg; $k_a$, $0.976hr^{-1}$; $k_{el}$, $0.104hr^{-1}$. The consistant kinetic parameters after a single and multiple oral administration show that there was no accumulation of rifapentine after 6 month oral administration. We also simulated the concentration of rifapentine after oral multiple administration of 10 and 50mg/kg/ day, based on the parameters obtained form the single administration. The measured serum concentrations of rifapentine were well fitted to the simulated results. The simulated results show that rifapentine readily reaches to steady-state after about 3 doses and the steady-state serum concentrations($C_{ss}$) are fluctuated in between $2.2{\sim}25.2{\mu}g/ml$, and $10.6{\sim}125.2{\mu}g/ml$ at the doses of 10 and 50mg/kg/day, respectively.

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A Tyrosinase Inhibitor Isolated from the Seeds of Eriobotrya japonica (비파 씨로부터 Tyrosinase 저해 활성물질의 분리)

  • Kim, Tae-Hoon;Shin, Seung-Ryeul;Kim, Tae-Wan;Lee, In-Chul;Park, Moon-Young;Jo, Cheo-Run
    • Food Science and Preservation
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    • v.16 no.3
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    • pp.435-441
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    • 2009
  • Activity-guided isolation from the ethylacetate (EtOAc)-soluble portion of a methanolic extract of the seeds of Eriobotrya japonica, using several bioassays, led to the isolation and identification of six phenolic compounds of previously known structure: benzaldehyde (1), chlorogenic acid (2), caffeic acid (3), benzoic acid (4), ferulic acid (5), and amygdalin (6). Of these, benzaldehyde (1) exhibited tyrosinase inhibitory activity in a bioassay. In addition, chlorogenic acid (2) and caffeic acid (3) were found to have strong antioxidative effects on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and superoxide dismutase (SOD)-like activity.

Inhibitory effect of by Ojeok-san lipid accumulation in high fat diet-induced obesity mice and 3T3-L1 adipocytes (오적산 (五積散)의 고지방식이 마우스 지방축적억제 및 3T3-L1지방세포에서의 비만 조절 기전에 관한 연구)

  • Choi, Hye-Min;Moon, Sung-Ok;Lee, Hee-Hyun;이, 화동
    • The Korea Journal of Herbology
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    • v.30 no.4
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    • pp.121-128
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    • 2015
  • Objectives : Ojeok-san (OJS), an oriental herbal formula, has been used in Asian countries including Korea, China and Japan to treat the common cold and illnesses including fatigue and gastrointestinal disorders. The purpose of this study was to examine the anti-obesity effect and molecular mechanism of OJS, on adipogenesis in 3T3-L1 cells. Also, the effects of OJS in obese mice fed a high-fat diet on adiposity were examined.Methods : Preferentially, we analyzed the component of OJS and measured the stability of its component in OJS according to study periods using high performance liquid chromatography (HPLC). In vitro, 3T3-L1 cells were treated with OJS (50 to 200 μg/mL) during differentiation for 8 days. The accumulation of lipid droplets was determined by Oil Red O staining. The expressions of genes related to adipogenesis were measured by RT-PCR and Western blot analyses. For anti-obesty effect in vivo, we experimented for 8 weeks with four group (normal diet (CON), high-fat diet (HF), high-fat diet with OJS (HF+OJS) and high-fat diet with Bang-pung-tong-sung-san (HF+BTS) in comparison group HF+OJS).Results : OJS showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affect cell toxicity as assessed by measuring fat accumulation and adipogenesis. In addition, OJS significantly reduced the expression levels of several adipocyte marker genes including proliferator activated receptor-γ(PPAR-γ) and CCAAT/enhancer-binding protein-α(C/EBP-α). Also OJS-administered mice showed significant inhibitory of body weights and abdominal adipose tissue weights.Conclusions : This study showed that traditional medicine OJS has an anti-obesity effect in vitro and in vivo. Thus, OJS could be developed as a supplement for reduction of body weight gain induced by an obesity.

Variation in Carotenoid Composition in Carrots during Storage and Cooking

  • Lim, Chae-Jin;Kim, Hyun-Young;Lee, Cheol-Ho;Kim, Yong-Ung;Back, Kyong-Whan;Bae, Jung-Myung;Lee, Shin-Woo;Ahn, Mi-Jeong
    • Preventive Nutrition and Food Science
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    • v.14 no.3
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    • pp.240-245
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    • 2009
  • High-performance liquid chromatography (HPLC) was applied to determine the carotenoid composition of carrots during storage and cooking. Analyses were conducted immediately after harvest and 1, 2, 4, and 8 weeks after harvest. During the course of the storage, the carotenoid levels generally decreased, and this decrease was found to be greater during the first week for $\beta$-carotene (all-trans-$\beta$-carotene) and lutein, and during the second week for $\alpha$-carotene. Additionally, the amount of the $\alpha-$ and $\beta$-carotenes in carrot leaves changed slightly within the first 2 weeks of harvest when stored at $4^{\circ}C$. Specifically, the level of lutein, the main component of carrot leaves, increased from 233.8$\pm$11.7 to $346.2\pm26.7{\mu}g$/g DW during the first 2 weeks. In addition, the change in carotenoid contents was observed during the home-processing of one Korean cultivar. Carrots fried in oil showed the highest amount of $\beta$-carotene ($164.3\pm6.6{\mu}g$/g DW) and $\alpha$-carotene ($50.1\pm0.4{\mu}g$/g DW), while carrots that were prepared by sauteing, pressure-cooking in water and microwaving had the second highest levels. The greatest loss of in carotenoids occurred in response to boiling in water containing 1% NaCl, braising and baking. The content of lutein increased slightly after boiling in water containing 1% NaCl ($9.3\pm0.4{\mu}g$/g DW), while a loss in lutein occurred after preparation using other home-processing methods. A cis-isomer of all-trans-$\beta$-carotene, 13-cis-$\beta$-carotene, was present in detectable amounts in all processed samples, but not in raw roots. Another isomer, 9-cis-$\beta$-carotene, was detected in carrots that were prepared by boiling, frying and pressure-cooking.

Influence of Ionic Liquid for Separation of D-tryptophan and N-CBZ-D-phenylalanine (D-tryptophan과 N-CBZ-D-phenylalanine의 분리에서 이온성 액체의 영향)

  • Jin, Yinzhe;Zheng, Jinzhu;Polyakova, Yulia;Koo, Yoon Mo;Row, Kyung Ho
    • Korean Chemical Engineering Research
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    • v.44 no.5
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    • pp.453-459
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    • 2006
  • D-tryptophan and N-CBZ-D-phenylalanine were separated using ionic liquid as additives for the mobile phase in high performance liquid chromatography (HPLC). The ionic liquid of 1-butyl-3- methylimidazolium tetrafluoroborate (${[BMIm]}^+{[BF_4]}^-$) was used. Mobile phases were 65%, 70%, and 80% methanol in water with addition of different concentrations (0.5, 1.0, 1.5, 2.0, 4.0, 6.0, 8.0, 10.0, 12.0, and 15.0 mmol/L) of the ionic liquid. The experiments were performed on stainless steel column, $3.9{\times}300mm$ i.d., packed with $15{\mu}m$ octadecyl-bonded silica gel at laboratory.The retention factor of D-tryptophan was not negligibly changed while that of N-CBZ-D-phenylalanine was decreased. The resolution between the two components were affected by the contents of methanol and ionic liquid in the mobile phase. With the small content of methanol and the high concentration of ionic liquid, the resolution was improved.

Effect of L-Ascorbic Acid Contents in Tissue on Collagen Synthesis in Guinea Pigs (Guinea pig의 조직중 L-Ascorbic acid함량이 콜라겐 생합성에 미치는 영향)

  • Yu, Rina;Kurata, Tadao;Arakawa, Nobuhiko
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.3
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    • pp.241-246
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    • 1992
  • To clarify the requirement of L-ascorbic acid (AsA) in collagen synthesis, the incorporation of 1-$^{14}$ C-proline into the tissues of guinea pigs and the specific radioactivity ratio (proline/hydroxyproline) in collagen were investigated. Male guinea pigs maintained on the AsA-deficient diet were divided into three groups ; group A (AsA-deficient animals) : group B (control animals) supplemented with 5mg AsA/day ; group C (high dose animals) with 300mg AsA/day, and orally supplemented with or with-out AsA for 14 days. Collagen synthesis was estimated by measuring the incorporation of labeled pro-line into collagen in lung and dorsal skin, and the hydroxyproline contents in lung and skin. The AsA contents in the tissues were determined by high-peforrnance liquid chromatography (HPLC), and serum alkaline phosphatase activity was also measured. The serum alkaline phosphatase activity of AsA deficient group was very low as compared with those of AsA supplemented group. Incorporation of labelled proline into collagen and its specific radioactivity ratio in collagen increased with increasing levels of AsA in the tissues. There was a significantly positive relationship between the levels of AsA and hydroxyproline in the tissues.

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The Effects of Sterilization on Jakyakgamcho Decoction(Shaoyaogancao Decoction) Pharmacopuncture: an Investigation to Minimize Loss of Principal Components (멸균에 따른 작약감초약침의 지표 성분변화 및 해결방안)

  • Lee, Jong Hwan;Jun, Jae Yun;Lim, Su Jin;Kim, Hae Sol;Kim, Ho Sun;Bae, Young Hyeon;Lee, In Hee;Kim, Min Jeong;Kim, Eun Jee;Ha, In Hyuk;Lee, Jin Ho;Lee, Jae Woong
    • Journal of Acupuncture Research
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    • v.31 no.4
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    • pp.29-32
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    • 2014
  • Objectives : Jakyakgamcho decoction is a traditional prescription known to be an effective pain control medication and muscle relaxant. For more localized treatment outcomes achieved in a shorter period of time, Jakyakgamcho decoction was reprocessed into a form of pharmacopuncture. An analysis of Jakyakgamcho decoction pharmacopuncture showed that there was a significant loss of paeoniflorin(Jakyak's index component). This study was designed to investigate ways to minimize this loss. Methods : After making changes to the processing methods of Jakyakgamcho decoction pharmacopuncture, we measured the quantity of paeoniflorin using high performance liquid chromatography(HPLC) for a before-and-after analysis Results : Paeoniflorin loss was observed 15 minutes after sterilization with $Na_2HPO_4$ at $121^{\circ}C$ Conclusions : It was found that paeoniflorin loss did not occur when pH was not controlled for during processing.