• Title/Summary/Keyword: hexane

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Time Course Variation of Vitamin $C_3$ Content in Leg Skin of Broiler Chicks Exposed to Different Dose of UVB Light (자외선의 상이한 선양을 조사한 브로일러 병아리의 다리 피부중 비타민 $C_3$ 함양의 경시적 변화)

  • 장윤환;김강수;여영수;강훈석;조인호;배은경
    • Korean Journal of Poultry Science
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    • v.20 no.2
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    • pp.93-105
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    • 1993
  • This study was carried out to determine the concentrations of previtamin D$_3$(PreD$_3$), lumisterol$_3$(L3), tachystero1$_3$(73), vitamin D$_3$(VD$_3$) and provitamin D$_3$(ProD$_3$) in leg skins of broiler chicks exposed to UVB lights (maximum intensity at 297 nm) with dose of 0.204 or 0.409 mJ/$\textrm{cm}^2$(30 or 60 min irradiation) . The broiler Hubbard line day old chicks(2 dose $\times$9 elapsed time $\times$4 replica+10 control=82) were fed VD-deficient diet for 31 days in a windowless subdued light room. The skin was collected at 0, 6, 12, 18, 30, 42, 66, 90 or 138 hr after UVB irradiation. The skin lipid was extracted by 9% ethyl acetate/n-hexane, and the fraction of VD$_3$ and its analogues was purified by Sep-Pak silica cartridge. The straight phase HPLC was utilized to analyze ProD$_3$ and its products. The mole %(absolute level expressed in ng/$\textrm{cm}^2$) of PreD$_3$ in leg skin (epidermis+dermis) was 4.67%(44 ng/$\textrm{cm}^2$) or 3.97%(37 ng/$\textrm{cm}^2$) right after UVB irradiation by 0.204 or 0.409 mJ/$\textrm{cm}^2$(30 or 60 min) at 15 cm distance, respectively. It content in leg skin at 0 hr after exposure was 7.24%(12 ng/$\textrm{cm}^2$) or 0.92%(9 ng/$\textrm{cm}^2$), respectively. The increase in irradiation dose did not affect proportionally the If synthesis.73 concentration in leg skin was 0.58%(S ng/$\textrm{cm}^2$) or 0.57%(6 ng/$\textrm{cm}^2$), respectively 0 hr after irradiation. The VD$_3$ in leg skin of birds exposed to UVB light with dose of 0.204 or 0.409 mJ/$\textrm{cm}^2$ was 2.13% (21 ng/$\textrm{cm}^2$) or 0.97% (16ng/$\textrm{cm}^2$), respectively at 0 hr after exposure, 2.72%(26ng/$\textrm{cm}^2$) or 3.84%(37ng/$\textrm{cm}^2$), respectively at 6 hr, and 4.30% ((33ng/$\textrm{cm}^2$) or 6.40%(76ng/$\textrm{cm}^2$), respectively at 12 hr. The peak concentration of VD$_3$ was presented at 18 or 30 hr when 0.204 or 0.409 mJ/$\textrm{cm}^2$) was treated, respectively. It was shown that 18~30 hr were necessary for the thermal conversion of PreD$_3$ into VD$_3$ in the leg skin of broiler chicks. The ProD$_3$ contents in leg skins of negative control, 0.204 mJ/$\textrm{cm}^2$ and 0.409 mJ/$\textrm{cm}^2$ treated birds were 966, 948 and 815 ng/$\textrm{cm}^2$, respectively at right before and after UVB exposure. It was estimated that 18 or 151 ng/$\textrm{cm}^2$ of ProD$_3$ was isomerized to PreD$_3$, L$_3$, T$_3$ and VD$_3$ when exposed to 0.204 or 0.409 mJ/$\textrm{cm}^2$, respective)y. Consequently it was shown that when double dose of UVB light was applied to irradiate the chick body, more but not double synthesis of VD$_3$ and its analogues was occured in leg skin of brolier chicks.

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Antioxidant and α-glucosidase inhibitory effects of ethanolic extract of Ainsliaea acerifolia and organic solvent-soluble fractions (단풍취 추출물 및 분획물의 항산화 및 α-glucosidase 저해 활성 평가)

  • Lee, Eun-Woo;Kim, Taewan;Kim, Hyun-Seok;Park, Youn-Moon;Kim, Seong-Ho;Im, Moo-Hyeog;Kwak, Jae Hoon;Kim, Tae Hoon
    • Food Science and Preservation
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    • v.22 no.2
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    • pp.275-280
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    • 2015
  • Among the naturally occurring antioxidants, polyphenols are widely distributed in various fruits, vegetables, wines, juices, and plant-based dietary sources and divided into several subclasses that included phenolic acid, flavonoids, stilbenes, and lignans. As part of our continuing search for bioactive food ingredients, the antioxidant and ${\alpha}$-glucosidase inhibitory activities of the aqueous ethanolic extract from the aerial parts of Ainsliaea acerifolia were investigated in vitro. The antioxidant properties were evaluated via radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) radicals. In addition, the anti-diabetic effect of A. acerifolia extracts was tested via ${\alpha}$-glucosidase inhibitory assay. Furthermore, the total phenolic contents were determined using a spectrophotometric method. All the tested samples showed dose-dependent radical scavenging and ${\alpha}$-glucosidase inhibitory activities. In particularly, the ${\alpha}$-glucosidase inhibitory and radical scavenging properties of the ethyl acetate (EtOAc)-soluble portion from the aerial parts of the A. acerifolia were higher than those of the other solvent-soluble portions. These results suggest that A. acerifolia could be considered a new potential source of natural antioxidants and antidiabetic ingredients. More systematic investigation of the aerial parts of A. acerifolia will be performed for the further development of anti-oxidative and antidiabetic drugs.

Phytochemical Analysis and Anti-cancer Investigation of Boswellia Serrata Bioactive Constituents In Vitro

  • Ahmed, Hanaa H;Abd-Rabou, Ahmed A;Hassan, Amal Z;Kotob, Soheir E
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7179-7188
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    • 2015
  • Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.

Isolation of Isoflavones and Soyasaponins from the Germ of Soybean (콩 배아로 부터 Isoflavone과 Soyasaponin의 동시 분리)

  • Kim, Sun-Lim;Lee, Jae-Eun;Kim, Yul-Ho;Jung, Gun-Ho;Kim, Dea-Wook;Lee, Choon-Ki;Kim, Mi-Jung;Kim, Jung-Tae;Lee, Yu-Young;Hwang, Tae-Young;Lee, Kwang-Sik;Kim, Wook-Han;Kwon, Young-Up;Kim, Hong-Sig;Chung, Ill-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.2
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    • pp.149-160
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    • 2013
  • The objective of present study was to simultaneously isolate of isoflavone and soyasaponin compounds from the germ of soybean seeds. Soy germ flours were defatted with hexane for 48h at room temperature, and methanolic extracts were prepared using reflux apparatus at $90^{\circ}C$ for 6h, two times. After extraction, extracts were separated with preparative RP-$C_{18}$ packing column ($125{\AA}$, $55-105{\mu}m$, $40{\times}150mm$), and collected 52 fractions were identified with TLC plate (Kieselgel 60 F-254) and HPLC, respectively. Among the identified isoflavone and soyasaponin fractions, isoflavone fractions were re-separated using a recycling HPLC with gel permeation column (Jaigel-W252, $20{\times}500mm$). Final fractions were air-dried, and the purified compounds of two isoflavones (ISF-1-1, ISF-1-2) and four soyasaponins (SAP-1, SAP-2, SAP-3, SAP-4) were obtained. Two isoflavone compounds (ISF-1-1, ISF-1-2) were acid-hydrolyzed for the identification of their aglycones, and confirmed by comparing with 12 types of isoflavone isomers. While the four kinds of soyasaponins were identified by using a micro Q-TOF mass spectrometer in the ESI positive mode with capillary voltage of 4.5kV, and dry temperature of $200^{\circ}C$. Base on the obtained results, it was conclude that ISF-1-1 is the mixture isomers of daidzin (43.4%), glycitin (47.0%), and genistin (9.6%), but ISF-1-2 is the single compound of genistin (99.8% <). On the other hand, soyasaponin SAP-1 is the mixture compounds of soyasaponin A-group (Aa, Ab, Ac, Ae, Af); SAP-2 is soyasaponin B-group (Ba, Bb, Bc) and E-group (Bd, Be); SAP-3 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$); SAP-4 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$, ${\beta}a$), respectively.

Anti-inflammatory and Whitening Effects of Protaetia brevitarsis Seulensis Extracts by Oriental Conversion Methods (포제방법에 따른 흰점박이꽃무지(Protaetia brevitarsis Seulensis) 추출물의 항염 및 미백활성에 관한 연구)

  • Sung, Gyeong Ah;Kim, Mi Hye;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.421-432
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    • 2016
  • In order to find new functional materials for the cosmetics application, we investigated anti-inflammatory and whitening effects of the Protaetia brevitarsis seulensis (P. brevitarsis) extracts, which were prepared by the various oriental conversion methods, as follows; fresh, roasted one time, roasted two times, roasted three times, and steamed. 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of the various solvent extracts (80% ethanol, 50% ethanol, ethyl acetate, hexane) of P. brevitarsis extracts were 85.5, 22.4, 37.0 and 19.4% respectively. The 80% ethanol extract with the highest antioxidant activity was used for all experiments. In case of antioxidant activity test of the extracts, all the extracts showed the activities in concentration dependent manner regardless of the sample preparation methods. Superoxide dismutase-like (SOD-like) activities of the extracts roasted three times and steamed were 62.9 and 55.9%, respectively in $500{\mu}g/mL$. Effects of extracts on the inflammation of RAW 264.7 cell induced by lipopolysaccharide (LPS) showed decreasing tendency of $NO{\cdot}$ and prostaglandin $E_2$ ($PGE_2$) production; PBS fresh (38.0%), PBS roasted one time (41.0%), PBS roasted two times (69.8%), PBS roasted three times (70.1%), PBS steamed (78.5%). Intracellular tyrosinase and melanin biosynthesis inhibitory activities of the extracts were decreased in a concentration dependent manner. However, the fresh P. brevitarsis extracts without the oriental conversion method showed 90.7% decrease compared to the control group treated with ${\alpha}$-MSH alone at $500{\mu}g/mL$. Taken together, these results suggest the oriental conversion method can be applied in development of cosmetic materials in order to improve anti-inflammatory and whitening effects of the cosmetics products.

Inhibitory Effects of Four Solvent Fractions of Alnus firma on α-Amylase and α-Glucosidase. (사방오리나무 추출물의 α-amylase 및 α-glucosidase 저해활성)

  • Choi, Hye-Jung;Jeong, Yong-Kee;Kang, Dae-Ook;Joo, Woo-Hong
    • Journal of Life Science
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    • v.18 no.7
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    • pp.1005-1010
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    • 2008
  • In this study, we investigated the inhibitory effect of four solvent fractions of Alnus firma on ${\alpha}-amylase$, ${\alpha}-glucosidase$ and aldose reductase activities. The inhibitory test showed that methanol (MeOH) extract and hexane (HX) fraction strongly inhibited pork pancreatin and salivary ${\alpha}-amylase$ activity. The MeOH extract and HX fraction of Alnus firma at the concentration of 4 mg/ml inhibited more than 70% of pancreatin and salivary ${\alpha}-amylase$ activity. The inhibitory effect of fractions has different specificities against ${\alpha}-amylase$ from pancreatin and salivary. In addition, the MeOH extract and butanol (BuOH) fraction showed the highest inhibitory activity on yeast ${\alpha}-glucosidase$ at values of $IC_{50}$ $137.36\;{\mu}g/ml$ and $115.14\;{\mu}g/ml$ respectively. The MeOH extract and BuOH fraction showed the highest inhibitory activity on yeast ${\alpha}-glucosidase$ than commercial agent such as 1-deoxynorjirimycin and acarbose. Inhibition kinetics of solvent fractions showed that ${\alpha}-glucosidase$ has been inhibited noncompetitively by the MeOH, EA and BuOH fraction. The aldose reductase from human muscle cell had been inhibited strongly by the MeOH extract and EA fraction at 57.996% and 83.293% at the concentration of $50\;{\mu}g/ml$, respectively. These findings may contribute to biological significance in that ${\alpha}-amylase$, ${\alpha}-glucosidase$ and aldose reductase inhibitory compounds could be used as a functional food and a drug for the symptomatic treatment of antidiabetic disease in the future.

Effects of Ethylacetate Fraction of Plantago asiatica L. on Hypercholesterolemia Induced by High Cholesterol Diet in Rats (질경이(Plantago asiatica L.) 에틸아세테이트분획이 흰쥐의 고콜레스테롤혈증에 미치는 효과)

  • Lee Jae-Joon;Lee Jeong-Hwa;Jeong Chang-Ju;Choi Hyun-Sook;Lee Myung-Yul
    • Food Science and Preservation
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    • v.12 no.6
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    • pp.624-630
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    • 2005
  • Plantain was extracted with ethanol and fractionated systemically with n-hexane, chloroform, ethylacetate, n-butanol and water to study inhibitory effect on cholesterol synthesis in vitro. To screen the effect, inhibitory activities on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase obtained from Saccharomyces cerevisiae were examined using the five fractions of Plantain. The HMG-CoA reductase activity was inhibited most by ehylacetate fraction among the fractions, although the all five fractions had the effect To see the hypocholesterolemic effect of the ethylacetate fraction of Plantanin (PAE) in vivo, male Sprague-Dawley rats were received 5 types of diets for 6 weeks: normal diet group (NOR), high cholesterol diet group($1\%$ cholesterol and $0.25\%$ sodium cholate, CON), normal diet and PAE 70 mg/kg administered group(S1), high cholesterol diet and PAE 140 mg/kg administrated group(S2), and high cholesterol diet and PAE 140 mg/kg administered group(S3). Body weight gains of the CON were significantly increased compared to those of S1, S2 and S3. Activities of serum AST and ALT were tended to be increased in CON compared with NOR and reduced by the PAE administration. Concentrations of serum total cholesterol, free cholesterol, LDL-cholesterol, triglyceride and the atherogenic index were tended to be decreased in the PAE administered groups compared with the CON. HDL-cholesterol and phospholipid concentrations were significantly decreased in the CON and markedly increased by the PAE administered groups. Taten together, it is suggested that the ethylacetate fraction of Plantanin exerts antiatherosclerotic effect by reducing serum cholesterol concentrations in rats fed high cholesterol diets.

Antimutagenic and Antitumor Effects of Codonopsis lanceolata Extracts (더덕 추출물의 항돌연변이 및 항종양 효과)

  • Kim, Soo-Hyun;Choi, Hyun-Jin;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.10
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    • pp.1295-1301
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    • 2009
  • This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effect of Codonopsis lanceolata (CL). CL was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of CL extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. CL extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of CL (200 ${\mu}g$/plate) showed approximately 72.1% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 69.6% and 67.0% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of CL extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (HepG2), human breast adenocarcinoma (MCF-7), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL CL ethyl acetate fraction had the highest cytotoxicity of 74.5%, 70.7% and 80.3% against HeLa, MCF-7 and A549 cells, respectively. In contrast, the extract and its fractions showed only 2$\sim$31% cytotoxicity for a normal human kidney cell line (293). In vivo anticancer effect of CL extract was tested using Balb/c mice transplanted sarcoma-180 cells. CL ethyl acetate fraction showed the highest inhibition rate of 56.4% at the 50 mg/kg concentration.

Determination of β-Carotene and Retinol in Korean Noodles and Bread Products (가공 및 외식식품 중 면류 및 제빵류의 레티놀 및 베타카로틴 함량 조사)

  • Shin, Jung-Ah;Chun, Ji Yeon;Lee, Junsoo;Shin, Ki Yong;Lee, Soon Kyu;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.12
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    • pp.1949-1957
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    • 2013
  • The contents of ${\beta}$-carotene and retinol in processed and restaurant foods, such as Korean noodles, mandus, rice cakes and bread products, were quantified by high-performance liquid chromatography (HPLC) with UV/visible and fluorescence detector, respectively. Samples were collected from different local areas (i.e. Gangwon-do, Gyeonggi-do, Gyeongsang-do, Seoul, Jeolla-do, and Chungcheong-do). After homogenization, samples were hydrolyzed by direct alkali saponification; thereafter, fat-soluble components were extracted by a mixture of n-hexane/ethylacetate (85:15, v/v), containing 0.01% butylated hydroxytoluene (BHT). ${\beta}$-carotene and retinol contents in infant formula used as an in-house material for the analytical quality control. Among 14 Korean noodles, high contents of ${\beta}$-carotene were found in Bibim-Guksu (average 442.43 ${\mu}g/100g$) and Jjolmyeon (average 301.39 ${\mu}g/100g$). In 4 Korean mandus, the highest contents of ${\beta}$-carotene was determined in Kimchi-mandu (average 197.64 ${\mu}g/100g$), resulting in 33.3 RE of the converted vitamin A. Among 12 Korean rice cakes, Maeun-Tteokbokki and Modm-Chaltteok contained relatively high content of ${\beta}$-carotene with 205.11 and 41.33 ${\mu}g/100g$, respectively, while retinol was detected only in Maeun- Tteokbokki (1.65~10.45 ${\mu}g/100g$). In addition, among 8 bread products, 77.3 RE of pastry, 51.2 RE of buttercream- bread, and 41.4 RE of morning roll were found as the contents of the converted vitamin A.

In vitro and In vivo Antimicrobial Activities of Medicinal Plants against Crown Gall in Grapevine (포도나무 줄기혹병균에 대한 약용식물의 항균활성 및 병발생억제)

  • Kim, Eun Su;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.34 no.4
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    • pp.537-548
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    • 2016
  • The objective of this study was to evaluate the antimicrobial activities of 9 kinds of medicinal plants against crown gall in grapevine. The medicinal plants extracted with several solvent systems were screened for in vitro antibacterial activity by the disc diffusion method. The ethanol and ethyl acetate extracts from magic lily flowers, tachys roots, asian plantain flowers and seeds, sweet wormwood leaves, stems and flowers, immature bitter melon fruits, cockscomb flowers, and peach tree resin showed in vitro antimicrobial activities against Rhizobium vitis with growth inhibition zones ranging from 10 to 27 mm in diameter. The minimum inhibitory concentration values of extracts against R.vitis ranged from 10,000 in Asian plantain flower and 50,000 fold diluted extracts in sweet wormwood flowers, stems, leaves, cockscomb leaves and immature bitter melon fruits. The active fractions of ethyl acetate and ethanol extracts from the medicinal plants were partially separated through silica gel column chromatography and thin layer chromatography (TLC). The active fractions were separated at Rf 0.36, 0.69, 0.75, 0.84, and 0.94 in sweet wormwood extracts, Rf 0.96 and 0.99 in cockscomb flower extracts, Rf 0.92 and 0.97 in cockscomb leaf extracts, and Rf 0.85 in immature bitter melon fruit extracts in TLC analysis developed with hexane:ethyl acetate (20:80, v/v) and methanol:chloroform (20:80, v/v). Among extracts from plants with in vitro antimicrobial activities, sweet wormwood, cockscomb leaves, and immature bitter melon fruits showed in vivo antimicrobial activities with inhibition activity of 100, 67, and 83.3%, respectively, in 'Kyoho' grapevine inoculated with R. vitis compared with the untreated control. These findings indicate that extracts of medicinal plants could be used as sustainable candidates to control crown gall disease caused by R. vitis in grapevines.