• Title/Summary/Keyword: Solvent Effect

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Collection, Identification and Hepatic Effect of Native Cordyceps militaris (새로운 번데기 동충하초의 수집, 동정 및 간기능에 미치는 효과)

  • Lee, Ki-Won;Nam, Byung-Hyouk;Jo, Wool-Soon;Oh, Su-Jung;Kang, Eun-Young;Cui, Yong;Lee, Jae-Yun;Cheon, Sang-Cheol;Jeong, Min-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.34 no.1
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    • pp.7-14
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    • 2006
  • Entomopathogenic fungus Cordyceps militaris is famous for its medicinal efficacies. It has been reported to have various pharmacological activities such as anti-tumour, insecticidal, antibacterial, immunomodulatory and antioxidant. In this study, we investigated the effect of the extract of C. militaris (MPUN8501), which was identified by the analysis of the nucleotide sequences of 5.8S ribosomal RNA, on the function of liver. C. militaris powder was extracted using hot water extracts method as time, volume and temperature and using method as differential polarity of organic solvent. Each fraction was tested for the improvement of hepatic enzyme alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity. The BuOH extracts (CME) had highest activity which was used for the test of toxicity and efficacy of C. militaris. The enhancing effect of CME on the activity of ADH and ALDH was much more than medicine, drink, natural tea etc. Thus CME promoted the resolution of alcohol and acetaldehyde in rats, inducing recovery to normal condition rapidly. Furthermore, oral administration of CME effectively protected the carbon tetrachloride-induced acute hepatic injury as revealed by the hematological parameters (levels of sGOT and sGPT) and histological observation. CME was ascertained to be safe by regulatory toxicity studies of single dose toxicity and genotoxicity. These results suggest that CME would be useful for the maintaining normal hepatic activity as a functional health food.

Effect of Pimpinella Brachycarpa Ethanol Extract on Chronically Ethanol-Induced Liver Damage in Rats (참나물이 만성적으로 알코올 유도된 흰쥐의 간손상에 미치는 영향)

  • Choo, Myung-Hee;Lee, Jae-Joon;Lee, Myung-Yul
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1406-1413
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    • 2007
  • This study was performed to investigate the effect of ethanol extract of Pimpinella brachycarpa(PBE) on chronically ethanol-induced hepatotoxicity in rat liver. Sprague-Dawley rats weighing 90-130 g were divided into 5 groups; normal group(NOR), ethanol(35%, 10 ml/kg) treated group(CON), PBE 200 mg/kg treated group(P1), PBE 200 mg/kg and ethanol treated group(P2), and PBE 400 mg/kg and ethanol treated group(P3). PBE was also fractionated by the following solvent: n-hexane, chloroform, ethylacetate and n-butanol. The antioxidative capacity of the n-hexane fraction was the highest among fractions and was similar to that of butylated hydroxytoluene(BHT). The body weight gain and feed intake of the rats were decreased by ethanol administration, but were gradually increased to the similar levels of the NOR group by administering PBE. The AST activity in serum elevated by ethanol was significantly decreased by administering the high dosage of PBE, but exerted no significant change on serum ALT activity. It was also observed that the hepatic activities of xanthine oxide(XO), catalase and glutathione peroxidase(GSH-Px) increased by ethanol were markedly decreased in the combined ethanol and PBE administered groups(P2 and P3), but not in the activity of superoxide dismutase(SOD) as compared with the CON group. The glutathione(GSH) contents were decreased by ethanol adminstration, however, increased after administering PBE. These results suggest that ethanol extract of Pimpinella brachycarpa has a possible positive effect on the liver function in hepatotoxicity-induced rats by ethanol administration.

Anti-oxidant and α-Glucosidase Inhibition Activity of Extracts or Fractions from Diospyros lotus L. Leaves and Quantitative Analysis of Their Flavonoid Compounds (고욤잎 추출물과 분획물의 항산화 및 alpha-glucosdiase 저해 활성 및 플라보노이드 화합물의 정량)

  • Kim, Seon-Young;Kim, Sang Jun;Kim, Ji-Ae;Kim, Da Hye;Kwak, Seol Hwa;Chung, Chang Ho;Jeon, In Hwa;Jang, Seon Il;Jeong, Seung-Il
    • Journal of Life Science
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    • v.24 no.9
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    • pp.935-945
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    • 2014
  • Persimmon leaves were commonly consumed as beverages, but were also used as popular folk medicine in Asia. The purpose of this work was to assess the biological activities of Diospyros Lotus L. extracts (DLLE). Various solvent extracts, including n-Hexnae, $CHCl_3$, EtOAc, and n-BuOH fractions, were obtained from the methanol extract of Diospyros Lotus L. leaves. The increasing interest in the powerful biological activity of plant phenolics and flavonoids outlined the necessity for determining their content in medicinal herbs. In this study, the total polyphenol and flavonoid contents (TPC and TFC) in the EA fraction were higher than those of other fractions. The biological activities of DLLE were tested using the 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity assay, as well as superoxide dismutase (SOD) activity as an anti-oxidant effect and ${\alpha}$-glucosidase inhibitory activity as an anti-diabetic effect. The EA fraction with high TPC and TFC values showed the highest anti-oxidant effect and high ${\alpha}$-glucosidase inhibition. The EA fractions were further purified into eight fractions using open column chromatography. Higher anti-oxidant and anti-${\alpha}$-glucosidase activity were observed in polar fractions. The content of the flavonoids, including quercein-3-O-rutinoside, kaempferol-3-O-glucoside, myricetin, luteolin, and kaempferol, were analyzed in effective fractions using high-performance liquid chromatography (HPLC). The results suggest that DLLE have anti-oxidative and anti-diabetic effects and thus, have the potential as anti-diabetic materials and as a source for natural health products.

Antimutagenic and Cytotoxicity Effects of Agaricus blazei Murill Extracts (아가리쿠스버섯(Agaricus blazei Murill) 추출물의 항돌연변이원성 및 세포독성 효과)

  • Ji, Jeong-Hwan;Kim, Mi-Nam;Choi, Kun-Pyo;Chung, Cha-Kwon;Ham, Seung-Shi
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1371-1378
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    • 2000
  • This study was performed to determine the antimutagenic and cytotoxic effect of Agaricus blazei Murill methanol extract on Salmonella typhimurium TA98, TA100 and human cancer cell lines using Ames test and cytotoxicity assay, respectively. In Ames test, methanol extract from A. blazei Murill did not exhibit any mutagenicity and most of the samples showed high antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), 4-nitroquinoline-1-oxide(4NQO), 3-amino-1,4-dimethyl-5H-pyrido [4,3-b] indol(Trp-P-1) and $benzo({\alpha})pyrene(B({\alpha})P)$. The methanol extracts of A. blazei Murill$(200\;{\mu}g/plate)$ showed approximately 92.4%, 81.9% and 83.4% inhibitory effect on the mutagenesis induced by 4NQO, Trp-P-1 and $B({\alpha})P$ against TA98 strain, whereas 87.3%, 94.7%. 92.3% and 89.9% inhibitions were observed on the mutagenesis induced by MNNG, 4NQO, Trp-P-1 and $B({\alpha})P$ against TA100 strain. The solvent fractions of methanol extracts from A. blazei Murill except water fraction showed high antimutagenic effects of $70{\sim}90%$ against mutation induced by MNNG, 4NQO. Trp-P-1 and $B({\alpha})P$. In anticancer effects of A. blazei Murill extract and fraction against cancer cell lines including human breast adenocarcinoma(MCF7), human lung carcinoma(A549), human fibrosarcoma(HT1080), human hepatocellular carcinoma(Hep3B), human epitheloid carcinoma(HeLa), human gastric carcinoma(KATO III) and human chronic myelogenous leukemia(K562) were investigated. The treatment of 1 mg/mL A. blazei Murill extracts had the highest cytotoxicity with 91.9% against HeLa, followed by KATO III(88.7%), A549(86.5%) and Hpe3B(84.3%). Whereas 1 mg/mL treatment of A. blazei Murill extracts had only $10{\sim}40%$ cytotoxicity on human normal liver cell (WRL68).

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STUDIES ON THE UTILIZATION OF ANTARCTIC KRILL 2. Processing of Paste Food, Protein Concentrate, Seasoned Dried Product, Powdered Seasoning, Meat Ball, and Snack (남대양산 크릴의 이용에 관한 연구)

  • PARK Yeung-Ho;LEE Eung-Ho;LEE Kang-Ho;PYEUN Jae-Hyeung;KIM Se-Kweun;KIM Dong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.13 no.2
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    • pp.65-80
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    • 1980
  • Processing conditions of the krill products such as paste food, krill protein concentrate, seasoned dried krill, powdered seasoning, meat ball, and snack have been examined and the quality was evaluated chemically and organoleptically. In the processing of paste food, krill juice was yielded $71\%$ and krill scrap $29\%$. The yields of paste and broth from the krill juice showed $53\%$ and $43\%$, respectively. In amino acid composition of the krill paste, proline, glutamic acid, aspartic acid, lysine, and leucine were abundant, while histidine, methionine, tyrosine, serine and threonine were poor. The optimum condition for solvent extraction in the processing of krill protein concentrate was the 5 times repetitive extraction using isopropyl alcohol at $80^{\circ}C$ for 5 mins. The yield of krill protein concentrate when used fresh frozen materials was $10.2\%$ in isopropyl alcohol solvent and $8.8\% in ethyl alcohol, and when used preboiled frozen materials, the yield was $13.0\%$ in isopropyl alcohol and $11.8\%$ in ethyl alcohol. Amino acid composition of krill protein concentrate showed a resemblance to that of fresh frozen krill meat. In quality comparison of the seasoned dried krill, hot air dried krill was excellent as raw materials and sun dried krill was slightly inferior to hot air dried krill, but preboiled frozen krill showed the poorest quality. The result of quality evaluation for seasoning made by combination of dried powdered krill, parched powdered sesame, salt, powdered beef extract, monosodium glutamate, powdered red pepper and ground pepper showed that the hot air dried krill was good in color and sundried krill was favorable in flavor. When krill meat ball was prepared using wheat flour, monosodium glutamate and salt as side materials, the quality of the products added up to $52\%$ of krill meat was good and the difference in quality upon the results of the organoleptic test for raw materials was not recognizable between fresh frozen and preboiled frozen krill. In the experiment for determining the proper amount of materials such as dried Powdered krill, $\alpha-starch$, sweet potato starch, sugar, salt, monosodium glutamate, glycine, potassium tartarate, ammonium bicarbonate, and sodium bicarbonate in processing krill snack, sample B(containing $7.7\%$ of dried powdered krill) and sampleC (containing $10.8\%$ of dried powdered krill) showed the most palatable taste from the view point of organoleptic test. Sweet potato starch in testing side materials was good in the comparison of suitability for processing krill snack. Corn starch and kudzu starch were slightly inferior to sweet potato starch, while wheat flour was not proper for processing the snack. In the experiment on frying method, oil frying showed better effect than salt frying and the suitable range of frying temperature was $210-215^{\circ}C$.

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Effect of 2-D DBP/PLGA Hybrid Films on Attachment and Proliferation of Intervertebral Disc Cells (2차원적 DBP/PLGA 하이브리드 필름이 디스크 세포의 부착과 증식에 미치는 영향)

  • Ko, Youn-Kyung;Jeong, Jae-Soo;Kim, Soon-Hee;Lim, Ji-Ye;Rhee, John-M.;Kim, Moon-Suk;Lee, Hai-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.2
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    • pp.109-115
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    • 2008
  • Because demineralized bone particle (DBP) contains various bioactive molecules such as cytokines, it is widely used biomaterials in the field of tissue engineering. In this study, we investigated the effect of 2-dimensional DBP/PLGA hybrid films on adhesion, proliferation and phenotype maintenance of intervertebral disc cells. PLGA films incorporated with different amount (0, 10, 20, 40 and 80 wt%) of DBP were prepared by the solvent evaporation method and characterized by scanning election microscopy (SEM). PLGA film has a flat and smooth surface. According to the increase of content of DBP, the surface of DBP/PLGA film exhibited few agglomerates and increased the roughness of the surface. Annulus fibrosus (AF) and nucleus pulposus (NP) cells were cultured on PLGA and DBP/PLGA film surface, and then examined the cell adhesion and proliferation by the cell count and SEM observation. The result of cell count and SEM observation revealed that 10 and 20% DBP in DBP/PLGA films were superior to adhesion and proliferation of both AF and NP cells. We confirmed that specific gene expression of disc cells on DBP/PLGA film based on the cell count result. Disc cells seeded on 20% DBP/PLGA film expressed the gene of type I and II collagen continuously. Therefore, pertinent content of biomaterials could provide more appropriate condition on adhesion and proliferation of cell. And this results may be used as a basic data for the intervertebral disc regeneration using tissue engineering.

An Analysis of a Porous Film Containing $Chamaecyparis$ $obtusa$ Extract (편백나무 추출물을 함유한 다공성 필름 분석)

  • Kim, Kyeong-Yee;Lee, Eun-Kyung
    • The Korean Journal of Food And Nutrition
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    • v.24 no.4
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    • pp.551-558
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    • 2011
  • This present study was performed to analyze the efficiency and volatility of a porous film containing $Chamaecyparis$ $obtusa$ extract as a method to effectively package food compounds. Phytoncide was contacted the state of gas and showed effective antimicrobial properties. Limonene can be distilled without decomposition as a relatively stable terpene and was one of the extract components. $Chamaecyparis$ $obtusa$ essential oil. The optimal solvent composition was a ratio 5:20:0.3 of T-500:ethanol:hardener to effectively manufacture film containing phytoncide essential oil and the minimum antibacterial concentration was 2%. The films were made under different conditions(A-50LF1, A-25SF2, B-50SF1, C-50LF1, C-25SF2 and D-50SF1) containing phytoncide and the amounts of limonene inside the 1-L reaction chamber depending on storage were measured by gas chromatography-mass selective detention. The results showed that the 25SF2(width, 25 mm; length, 20 cm) revealed more amount of limonene compared with 50LF1(width 50 mm, length 20 cm). We confirmed that the gas emission amount showed a better layer on the film side than on the internal film. An effect of film thickness on phytoncide emissions was observed in that the amounts was less than the expectation for a thicker film at the beginning time, but the emitting amounts increased with increasing storage periods. In the storage testing of various films at $35^{\circ}C$ and 70% humidity for 14 days, 25SF2 showed longer preservation compared with that of 50LF in the case of bread. $C.$ $obtusa$ essential oil is a useful fresh ingredients, hence, analysis of limonene emission kinetics from various film was helpful to develop films with an optimal antimicrobial effect, and will allow application of such films in food packaging systems.

Effects of Aqueous Azadirachta indica Extract on Hepatotoxicity in Rats (수용성 님추출물이 랫드의 간 독성에 미치는 영향)

  • Park, Kyung-Hun;Yoon, Hyunjoo;Han, Beom Seok;Lee, Je-Bong;Jeong, Mi Hye;Cho, Namjun;Om, Ae Son;Paik, Min-Kyoung
    • Korean Journal of Environmental Agriculture
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    • v.33 no.4
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    • pp.395-402
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    • 2014
  • BACKGROUND: Azadirachta indica Extract(AIE) containing azadirachtin as active ingredient have been used worldwide as environment-friendly organic material having pest control properties. However, the extracts prepared with different solvent and from different plant site is very diverse and have different toxicity. METHODS AND RESULTS: In this study, the four week repeated oral dose toxicity test of aqueous AIE in Sprague-Dawley rats was carried out to investigate the toxic effect of liver, main toxicity target organ of AIE. The male and female rats were divided into 4 groups, respectively; control(0 g/Kg bw), low-dose group(0.5 g/Kg bw), middle-dose(1.0 g/Kg bw) and high-dose group(2.0 g/Kg bw). As a results, relative liver weight increased with dose-dependent of AIE(p<0.05). Serum LDH in all AIE-treated groups were significantly lower than the control in male rats(p<0.05). However, serum GOT and GPT were significantly increased in all male AIE-treated groups in male rats(p<0.05) and, in particular, increase of serum GPT in dose-dependent manner raise the possibility of liver damage. Even through serum GLU was increased significantly in high-dose group in male rats compared to control, there were no significant differences of urinary GLU among all groups(p<0.05). In addition, histopathological examination of the liver did not reveal any lesions in all AIE-treated groups. CONCLUSION: In conclusion, 4 weeks of the repeated oral administration of AIE 2.0 g/Kg to rats has resulted no toxic response in liver. Therefore, AIE was no indicated to have any toxic effect in the SD rats, when it was orally administrated below the dosage 2.0 g/Kg/day for 4weeks.

Free Radical Scavenging Effect and Extraction Condition of Ethanol Extracts of Epimedium Koreanum Nakai Containing Different Icariin Quantity (Icariin 함량에 따른 삼지구엽초 추출물의 추출조건 및 항산화 효과)

  • Kim, Seo-Jin;Oh, Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.359-364
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    • 2007
  • This study was conducted to determine the optimum condition of extraction and antioxidant activity of ethanol extracts of Epimedium koreanum Nakai based on the icariin quantity. Also, further organic solvent fractions of hexane, chloroform, ethyl acetate and butanol were obtained from the ethanol extract of Epimedium koreanum Nakai at different temperatures. Total ethanol extraction yield of wild grape seed ranged from 11.8% to 39.3% depending on the concentration of icariin as well as different ethanol concentration, extraction temperature and time condition. Among different extraction temperatures and time, the highest extraction yield of 39.3% was obtained at 70% ethanol for 3 hour at $50^{\circ}C$ in the sample containing the 0.596% icariin (KE9412). In the meantime, the strongest free radical scavenging effect $(RC_{50})$ with $18.9{\mu}g/mL$ in the KE9412 sample was observed in 70% ethanol extract of Epimedium koreanum Nakai extracted for 7 hour at $70^{\circ}C$, while $RC_{50}$ with $35.2{\mu}g/mL$ was observed in the KE9405 (0.20% icariin content) sample at same condition. Also, antioxidant activity of ethanol extracts of Epimedium koreanum Nakai increased as icariin concentration increased. Among each fraction obtained from organic solvents, butanol fraction was found to have the strongest $RC_{50}\;(39.2{\mu}g/mL)$ and followed by ethylacetate $(49.0{\mu}g/mL)$, water $(118.8{\mu}g/mL)$, ethylacetate $(119.8{\mu}g/mL)$, and chloroform fraction $(138.5{\mu}g/mL)$ respectively.