• Title/Summary/Keyword: spectrophotometric assay

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Acetylcholinesterase Inhibitory and Antioxidant Properties of Aster yomena Extract (쑥부쟁이 추출물의 아세틸콜린에스테라제 저해 및 항산화 활성)

  • Bae, Jong-Sup;Kim, Tae-Hoon
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.121-126
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    • 2009
  • Objectives : To evaluate the radical scavenging and acetylcholinesterase (AChE) inhibitory activities of the ethylacetate (EtOAc)-soluble portion of a methanolic extract of Aster yomena, three different assay systems were performed. Methods : The antioxidant activity of A. yomena extract was tested as its capacity to scavenging free radicals of DPPH and $ABTS^+$, which has been widely used to evaluate the antioxidant activity of natural products from plant sources. AChE inhibitory activity was tested against mouse brain AChE by spectrophotometric method of Ellman using ELISA microplate reader. Results : The methanolic extract of A. yomena was fractionated and the EtOAc-soluble portion showed significant AChE inhibitory and free radical scavenging effects. Also the EtOAc-soluble portion revealed the highest phenolic contents as compared to the other extracts. Conclusions : These results indicate that phenolic compounds may be important constituents that give rise to the anti-AChE and antioxidative activities of A. yomena extract. Further phytochemical studies on this plant, for nutraceutical or pharmaceutical application, are warranted.

Transition Temperature of Mitochondrial Respiratory Activity as an Index for Chilling Susceptivity of Varieties of Rice Plant (미토콘드리아 호흡활성 전이온도의 측정에 의한 벼품종별 상대적 내냉성판정)

  • Jung, Jin;Kim, Hee-Jung
    • Applied Biological Chemistry
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    • v.30 no.4
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    • pp.357-363
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    • 1987
  • The transition temperature of respiratory activity of mitochondria isolated from panicles of rice plant was measured with 41 varieties bred in Korea, which had been agronomically classified into five groups based on their relative cold-tolerance; S (strong cold-tolerance), S-M (moderate to strong cold-tolerance), M (moderate cold-tolerance), M-W (moderate to weak cold-tolerance) and W (weak cold-tolerance). The temperature was found to be $11.5{\sim}13.5^{\circ}C$ for group S, $13.5{\sim}16^{\circ}C$ for group S-M, $16{\sim}17^{\circ}C$ for group M, $17{\sim}18^{\circ}C$ for group M-W, and ${\geq}18^{\circ}C$ for group W respectively with a few exceptions of minor deviation, demonstrating a close correlation between the transition temperature and the chilling susceptivity of a variety. This observation supports the suggestion that the transition temperature of respiratory activity of panicles be adopted as a reliable index for grading a rice variety as to its overall cold tolerance. Also studied is on the methodology for measurement of the transition temperature in the aspect of analytical biochemistry. In addition to the usual polarographic method, a spectrophotometric method which is based on the enzymatic assay of membrane-bound cytochrome C oxidase was examined. This enzyme was found previousely and confirmed again in the present work to be a key factor governing the respiratory trantion at least in plant mitochondria. The optical method was turned out to be very successful, although it was a little more complicate and time-consuming than the electrochemical method.

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Comparative Analysis of Melanin Contents in Hairs among Cattle Breeds (소의 모발 멜라닌 색소 함량의 품종별 비교 분석)

  • Lee, S.S.;Kang, S.R.;Han, S.H.;Cho, I.C.;Shin, K.Y.;Cho, Y.I.;Kang, T.Y.;Ko, Moon-Suck;Yang, Y.H.
    • Journal of Animal Science and Technology
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    • v.49 no.1
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    • pp.147-154
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    • 2007
  • To characterize the colorization patterns of bovine hairs, the melanin contents were quantitatively assayed and compared among cattle breeds. The total melanin levels measured by spectrophotometric assay (A500) from Jeju Black cattle were significantly lower than those from Holstein or Angus with black coat color but significantly higher than those from Hanwoo with yellow coat color or Angus and Holstein with red coat color (P<0.001). The total melanin levels from Hanwoo were significantly lower than those from Red Angus and Red Holstein but significantly higher than those from Hanwoo×Charolais crossbred (P<0.001). The relative ratios of eumelanin to pheomelanin (A650/A500) were 0.382, 0.359, 0.112 and 0.124 in Angus, Jeju Balck cattle, Red Holstein and Hanwoo, respectively. These results show that the spectrophotometric method provides a convenient way to qualitatively characterize hair melanin and may be useful for studying expression of major coat color genes in Hanwoo and Jeju Black cattle.

Determination of Selected Antioxidant Compounds in Specialty Rice (여러 가지 특수미의 일부 항산화 성분 분석)

  • Seo, Sun-Jung;Choi, Young-Min;Lee, Sun-Mi;Kim, Kee-Jong;Son, Jong-Rok;Lee, Jun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.4
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    • pp.499-502
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    • 2007
  • The objective of this study was to determine antioxidant compounds in specialty rice including milled rice, brown rice, red rice, giant embryonic rice, black rice, green rice, and Goami consumed in Korea. The concentrations of total polyphenols (insoluble and soluble polyphenols), phytic acid, and anthocyanin in the samples were measured using spectrophotometric methods and vitamin I analysis was carried out by HPLC. The contents of the total polyphenolic compounds were 565mg/100g for black rice, 405mg/100g for red rice, 140mg/100g for giant embryo rice, 138mg/100g for Goami, 133mg/100g for brown rice, 127mg/100g for green rice, and 66mg/100g for white rice. The black and red rices were significantly high in polyphenolic contents compared with the other rices, apparently due to their intense red-purple color. Black rice, red rice, and Goami showed significantly higher vitamin E and phytic acid contents compared with other rices. Anthocyanins were determined in only black rice (302mg/100g) due to the detection limits of spectrophotometric assay. hlthough vitamin I and anthocyanin contents were relatively lower than polyphenolics among the samples, the specialty rice may contribute to the significant supply of antioxidant compounds to prevent oxidative stress due to the fact that rice is used as a staple food and consumed in large amounts in our diets. The results can be used to increase rice consumption by enhancing consumer awareness on health benefits of the rice.

Spectrophotometric Determination of Aminoglycoside Antibiotics Based on their Oxidation by Potassium Permanganate (과망간산포타슘에 의한 산화에 바탕을 둔 아미노글리코사이드 항생제의 분광광도법적 정량)

  • El-Didamony, A. M.;Ghoneim, A. K.;Amin, A. S.;Telebany, A. M.
    • Journal of the Korean Chemical Society
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    • v.50 no.4
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    • pp.298-306
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    • 2006
  • A rapid, simple and sensitive validated spectrophotometric methods have been described for the assay of neomycin and streptomycin either in pure form or in pharmaceutical formulations. The proposed methods were based on the oxidation of the studied drugs by a known excess of potassium permanganate in acidic medium and estimating the unreacted permanganate with amaranth dye (method A), acid orange II (method B), indigocarmine (method C), and methylene blue (method D), in the same acid medium at a suitable lmax=521, 485, 610 and 664 nm, respectively. Beers law is obeyed in the concentration range of 5-10 and 2-7 mg mL-1 for neomycin and streptomycin, respectively. The apparent molar absorptivity and sandell sensitivity values are in the range 5.47-6.20104, 2.35-2.91105 L mol-1 cm-1 and 7.57-8.59, 5.01-6.2 ng cm-2 for neomycin and streptomycin, respectively. Different variables affecting the reaction were studied and optimized. The proposed methods were applied successfully to the determination of the examined drugs either in a pure or pharmaceutical dosage forms with good accuracy and precision. No interferences were observed from excipients and the results obtained were in good agreement with those obtained using the official methods.

Determination of Bioactive Compounds and Anti-cancer Effect from Extracts of Korean Cabbage and Cabbage (배추와 양배추 추출물의 생리활성 물질 및 암세포 증식 억제효과 분석)

  • Hwang, Eun-Sun;Hong, Eun-Young;Kim, Gun-Hee
    • The Korean Journal of Food And Nutrition
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    • v.25 no.2
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    • pp.259-265
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    • 2012
  • In this study, we determined total polyphenol content(TPC) and total flavonoid content(TFC) of extracts from Korean cabbage and cabbage using a spectrophotometric method as well as glucosinolates concentration by HPLC. TPCs of Korean cabbage and cabbage extracts were 308.48 ${\mu}g$ GAE/g dry weight and 344.75 ${\mu}g$ GAE/g dry weight, respectively. TFCs of Korean cabbage and cabbage extracts were 5.33 ${\mu}g$ QE/g dry weight and 5.95 ${\mu}g$ QE/g dry weight, respectively. We found six different glucosinolates, namely progoitrin, glucoalyssin, gluconapin, glucobrassicanapin, glucobrassicin and 4-methoxyglucobrassicin in the Korean cabbage extract. In the cabbage extract, there was four glucosinolates, namely glucoraphanin, sinigrin, glucobrassicin and 4-methoxyglucobrassicin. We determined the cytotoxic effect of Korean cabbage and cabbage extracts in AGS human stomach cancer cells, HepG2 human hepatic cancer cells and LNCaP human prostate cancer cells by MTT assay. Dose-dependent relationships were found between the extract concentrations and cancer cell growth inhibition. The overall results support that both Korean cabbage and cabbage, the major vegetables in Korea, contain bioactive compounds such as polypheol, flavonoids as well as glucosinolates and they may play a positive role in cancer prevention.

Development of Axenic Culture and Astaxanthin Production in Microalgae (미세조류를 이용한 무균분리법 개발 및 astaxanthin 생산)

  • Son, Min Chang;Lee, Dong-Jun;Park, Sejin;Kim, Min Sung;Lee, Chul Won;An, Won Gun
    • Journal of Life Science
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    • v.25 no.7
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    • pp.733-739
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    • 2015
  • Microalgae are a renewable natural resource that requires only sunlight, carbon dioxide, phosphorus, and nitrogen for rapid growth. They produce a broad variety of basic chemical substances―such as vitamins, fatty acids and carotenoids―that have high added value potential for the pharmaceutical and food industries. The aim of this study was to develop axenic culture and to establish a cell growth assay for microalgae. A further experiment was carried out to determine the yield of astaxanthin derived from microalgae. The axenic culture was developed using a mixture of antibiotics [ampicillin (100 ${\mu}g/ml$), streptomycin (10 ${\mu}g/ml$), chloramphenicol (10 ${\mu}g/ml$), penicillin (10 ${\mu}g/ml$), neomycin (50 ${\mu}g/ml$), gentamycin (50 ${\mu}g/ml$), kanamycin (10 ${\mu}g/ml$), and nystatin (1.5 ${\mu}g/ml$)] and then used to extract a variety of useful components from the microalgae. The optimal concentration for the antibiotic mixture was 1-3 percent. A spectrophotometric cell growth assay was also established. Astaxanthin was extracted from Haematococus lacustris with a yield of $1.9{\times}10^{-3}{\mu}g/l$ per 1 ml of culture medium. In conclusion, the axenic culture method developed here allows extraction of high-quality astaxanthin and other useful components from microalgae.

Highly Active Analogs of α-Factor and Their Activities Against Saccharomyces cerevisiae

  • Ahn, Hee Jun;Hong, Eun Young;Jin, Dong Hoon;Hong, Nam Joo
    • Bulletin of the Korean Chemical Society
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    • v.35 no.5
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    • pp.1365-1374
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    • 2014
  • Thirteen analogs of tridecapeptide ${\alpha}$-factor (WHWLQLKPGQPMY) of Saccharomyces cerevisiae with C- or N-terminal Trp extension and isosteric replacement by Aib at position 8 and 11, Trp at position 13, D-Ala at position 9, and Orn and Glu at position 6 were synthesized and assayed for their biological activity. Receptor binding assay was carried out using our newly developed spectrophotometric method with detector peptide 14. C- or N-terminal extended analogs, ${\alpha}$-factor-$[Trp]_n$ (n =1-5) 1-5 and $[N-Trp]_1$-${\alpha}$-factor 6, were all less active than native ${\alpha}$-factor and gradual decreases in both activity and receptor affinity were observed with greater Trp extension. Trp-substituted analog at position 13, $[Trp^{13}]{\alpha}$-factor 7, exhibited about 2-fold reductions in both activity and receptor affinity. Aib-substituted analogs, $[Aib^8]{\alpha}$-factor 8 and $[Aib^{11}]{\alpha}$-factor 9, showed 5- to 10-fold reduction in activity as well as 3-fold reduction in receptor affinity compared to native ${\alpha}$-factor. $[Orn^6]{\alpha}$-factor 10 demonstrated strong potency with a 7.0-fold increase in halo activity as well as 1.8-fold increase in receptor affinity compared to native ${\alpha}$-factor. For two double substituted analogs, [$Glu^6,{\small{D}}-Ala^9$]${\alpha}$-factor 12 showed the slightly decreased potency in halo activity compared to analog 10, whereas [$Orn^6,{\small{D}}-Ala^9$]${\alpha}$-factor 11 exhibited 15-fold higher halo activity as well as nearly 3-fold higher receptor affinity compared to native ${\alpha}$-factor.

Study of Hair Melanins in Various Hair Color Alpaca (Lama Pacos)

  • Fan, Ruiwen;Yang, Gang;Dong, Changsheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.4
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    • pp.444-449
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    • 2010
  • The aim of this study was to measure the hair melanins of various colors and to find the relationship between the quantity of melanins and hair color phenotypes in alpacas. According to the Munsell color system, 3 healthy alpacas were selected for each of the 22 different hair color phenotypes (66 alpacas altogether). Alpaca hair was taken from the lateral thoracic region and then dissolved with different solutions to obtain melanins. The values of alkali-soluble melanins (ASM), eumelanin (EM) and pheomelanin (PM) were measured by spectrophotometric assay, and labeled as Sp.ASM, Sp.EM and Sp.PM, respectively. Data were analyzed using SPSS11.5 software. Results showed that average Sp.ASM and Sp.PM were increased as the color deepened from white to black, ranging from 0.500 to 4.543 for Sp.ASM and from 0.268 to 1.457 for Sp.EM. However, average Sp.PM had no such apparent relationship with color. Based on the value of Sp.ASM and EM, 7 hues were produced and gray was a single hue. Most of the data were in a normal distribution (p>0.10). ANOVA analysis showed that mean values of Sp.ASM, Sp.EM and Sp.PM were significantly different (p<0.05). The results also showed that Sp.ASM was positively correlated with Sp.EM but the correlation between Sp.ASM and Sp.PM was not significantly different from 0. It is concluded that EM is the major constituent of alpaca hair melanin; there is a significant correlation among ASM, EM and alpaca hair colors, and EM is the most reliable parameter for distinguishing these groups.

Biochemical Characteristics and Function of a Fucosyltransferase Encoded by ste7 in Ebosin Biosynthesis of Streptomyces sp. 139

  • Chang, Ming;Bai, Li-Ping;Shan, Jung-Jie;Jiang, Rong;Zhang, Yang;Guo, Lian-Hong;Zhang, Ren;Li, Yuan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1092-1097
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    • 2009
  • A novel exopolysaccharide named Ebosin was produced by Streptomyces sp. 139, with medicinal activity. Its biosynthesis gene cluster (ste) has been previously identified. For the functional study of the ste7 gene in Ebosin biosynthesis, it was disrupted with a double crossover via homologous recombination. The monosaccharide composition of EPS-7m produced by the mutant strain Streptomyces sp. 139 ($ste7^-$) was found altered from that of Ebosin, with fucose decreasing remarkably. For biochemical characterization of Ste7, the ste7 gene was cloned and expressed in Escherichia coli BL21. With a continuous coupled spectrophotometric assay, Ste7 was demonstrated to have the ability of catalyzing the transfer of fucose specifically from GDP-$\beta$-L-fucose to a fucose acceptor, the lipid carrier located in the cytoplasmic membrane of Streptomyces sp. 139 ($ste7^-$). Therefore, the ste7 gene has been identified to code for a fucosyltransferase, which plays an essential role in the formation of repeating sugars units during Ebosin biosynthesis.