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Simultaneous Determination of Vitamin A and E in Infant Formula by HPLC with Photodiode Array Detection

  • Lee, Hong-Min;Kwak, Byung-Man;Ahn, Jang-Hyuk;Jeong, Seung-Hwan;Shim, Sung-Lye;Kim, Kyong-Su;Yoon, Tae-Hyung;Leem, Dong-Gil;Jeong, Ja-Young
    • Food Science of Animal Resources
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    • v.31 no.2
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    • pp.191-199
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    • 2011
  • The objective of this study was to develop a method to simultaneously quantify vitamins A and E in infant formula. To determine the vitamin A and E content, vitamin A and four different vitamin E isomers (${\alpha}$-, ${\beta}$-, ${\gamma}$-, and ${\delta}$-tocopherol) were separated by high performance liquid chromatography with a photodiode array detector using a Develosil RPAQUEOUS RP-$C_{30}$ column ($4.6{\times}250$ mm, 5 ${\mu}M$). The vitamin A and E contents in the certified reference material determined using this method were within the certified range of standard values. The limits of detection (LODs) and limits of quantitation (LOQs) for vitamin A were 0.02 and 0.06 ${\mu}g/L$, respectively. LODs and LOQs for the vitamin E isomers ranged from 0.20 to 0.55 and from 0.67 to 1.81 ${\mu}g/L$, respectively. Linear analyses indicated that the square of the correlation coefficient for the vitamin A and E isomers was 0.9997-0.9999. The recovery of vitamins ranged from 96.69 to 97.79%. The results demonstrate that this novel method could be used to reliably analyze vitamin A and E content in infant formula.

A New Detergentless Micro-Emulsion System Using Urushiol as an Enzyme Reaction System

  • Kim, John-Woo-Shik;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.369-375
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    • 2001
  • Urushiol, a natural monomeric oil, was used to prepare a detergentless micro-emulsion with water and 2-propanol The formation of micro-emulsion was verified by conductivity measurements and dynamic light scattering. The conductivity data showed phase change dynamics, a characteristics of micro-emulsions, and subsequent dynamic light scattering study further confirmed the phenomenon. Average water droplet diameter was 10 nm to 500 nm when the molar ratio of 2-propanol ranged from 0.40 to 0.44 . Earlier studies were performed on toluene and hexane, in which the insoluble substrate in water phase was added to the solvents to be reacted on by enzymes. However, in the present urushiol system, urushiol was used as both solvent and substrate in the laccase polymerization of urushiol. The laccase activity in the system was examined using polymerization of urushiol. The laccase activity in the system was examined using syringaldezine as a substrate, and the activity increased rapidly near the molar ratio of 2-propanol at 0.4, where micro-emulsion started. The activity rose until 0.46 and fell dramatically thereafter. The study of laccase activity in differing mole fractions of 2-propanol showed the existence of an ‘optimal zone’, where the activity of laccase was significantly higher. In order to analyze urushiol polymerization by laccase, a bubble column reactor using a detergentless micro-emulsion system was constructed. Comparative study using other organic solvents systems were conducted and the 2-propanol system was shown to yield the highest polymerization level. The study of laccase activity at a differing mole fraction of 2-propanol showed the existence of an ‘optimal zone’ where the activity was significantly higher. Also, 3,000 cP viscosity was achieved in actual urushi processing, using only 1/100 level of laccase present in urushi.

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Quality Evaluation of Moutan Cortex Radicis Using Multiple Component Analysisby High Performance Liquid Chromatography

  • Ding, Yan;Wu, Enqi;Chen, Jianbo;Nguyen, Huu-Tung;Do, Thi-Ha;Park, Kyung-Lae;Bae, Ki-Hwan;Kim, Young-Ho;Kang, Jong-Seong
    • Bulletin of the Korean Chemical Society
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    • v.30 no.10
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    • pp.2240-2244
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    • 2009
  • A simple high performance liquid chromatographic method was developed to evaluate the quality of Moutan Cortex Radicis based on chromatographic fingerprints that characterize eight pharmacological compounds, namely, gallic acid, paeoniflorin, galloyl paeoniflorin, benzoic acid, quercetin, benzoylpaeoniflorin, paeoniflorigenone, and paeonol. These compounds were identified by their characteristic UV profiles and the mass spectroscopy data, and their contents were determined by HPLC. The chromatographic separation was performed on a $C_{18}$ column by gradient elution with 0.05% formic acid in water and acetonitrile. The methodological validation gave acceptable linearities (r = 0.9996) and recoveries (ranging from 99.4∼103.1%). The limits of detection (LOD) of these compounds ranged from 10 to 30 $\mu$g/mL. The representative chromatographic fingerprints of Moutan Cortex Radicis were obtained by analyzing 20 batches of samples collected from markets in Korea and China. For the efficient evaluation of quality for the commercial Moutan Cortex Radicis it is recommended that the total content of the six characteristic compounds should contain more than a minimum of 2% and that the content of total paeoniflorin and paeonol should exceed a minimum of 1.5% of dry weight of Moutan Cortex Radicis.

Hygienic Studies on Inactive Substances of Sterigmatocystin by Artificial Gastric Juice (인공(人工) 위액(胃液)에 의한 Sterigmatocystin의 실활물(失活物)에 관한 위생화학적(衛生化學的) 연구(硏究))

  • Kim, Soo-Yeon;Oh, You-Jin;Yeo, Sin-Koo;Jang, Seong-Jae
    • The Korean Journal of Mycology
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    • v.14 no.1
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    • pp.61-70
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    • 1986
  • Sterigmatocystin bears a close structural relationship to aflatoxin $B_1$ and is a carcinogenic compound that has been shown to affect various species of experimental animals. Reaction and toxicity of sterigmatocystin in the artificial gastric juice were investigated. Sterigmatocystin was degraded in artificial gastric juice and extracted by the method of A.O.A.C. After cleaned up by silica gel column chromatography, this substance was detected and characterized by thin layer chromatography, UV, IR and mass spectra. It showed $R{\mathcal{f}}$ 0.4 and brick-red color by TLC. Especially, in the mass spectrum of it, fragment peak at m/e 327 was due to the loss of the $-CH_3$ and $-H_2O$, fragment peak at m/e 341 was due to the loss of the $H_2O$ and $-H^+$, and fragment peak at m/e 239 was due to the loss of the 2-chloro-tetrahydrofuran and methyl group from the parent molecule. Therefore, a degraded substance of sterigmatocystin reacted in artificial gastric juice (Sub. K) was estimated with additional formation of hydrochloric acid. In four-day-old chicken embryos, the mean lethal dose $(LD_{50})$ was $140\;{\mu}g/egg$, and 90 to 100% of the embryos were killed with 1 mg/egg. This $LD_{50}$ $140\;{\mu}g/egg$ compared with an $LD_{50}$ $14.69\;{\mu}g/egg$ for sterigmatocystin (acute toxicity) showed the substance to be much less toxic than sterigmatocystin.

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Ovicidal Activity of Lactic Acid Produced by Lysobacter capsici YS1215 on Eggs of Root-Knot Nematode, Meloidogyne incognita

  • Lee, Yong Seong;Naning, Kyaw Wai;Nguyen, Xuan Hoa;Kim, Sun Bae;Moon, Jae Hak;Kim, Kil Yong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1510-1515
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    • 2014
  • Lysobacter capsici YS1215 isolated from soil previously showed nematicidal potential for biological control of the root-knot nematode. In this study, lactic acid, a nematicidal compound, was isolated from culture filtrate of YS1215, and its ovicidal activity was investigated. Purification and identification of lactic acid were performed by a series of column chromatographies and identified by $^1H$ and $^{13}C$ NMR spectra and GC-MS analysis. Our results showed that bacterial culture filtrate containing lactic acid significantly inhibited egg hatching. The lowest egg hatch rate (5.9%) was found at a high concentration ($25 {\mu}l/ml$) of lactic acid at 5 days after incubation, followed by 20 (15.2%), 15 (23.7%), 10 (29.8%), and $5(36.4%){\mu}l/ml$, while egg hatching in the control (sterile distilled water) was 44.5%. This is the first report of lactic acid as an ovicidal compound, and it may be considered as an alternative of chemical pesticide against root-knot nematodes.

Beta-carotene and Lutein Contents in Green Leafy Vegetables (녹색잎 채소류의 ${\beta}$-Carotene과 Lutein 함량)

  • Lee, Hwa-Suk;Kim, Young-Nam
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.2
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    • pp.175-180
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    • 1997
  • Beta-carotene and lutein contents in 7 different green leafy vegetables(perilla leaf, mugwort, chwi, lettuce, spinach, leek, and crown daisy) were analyzed by HPLC. The isocratic separation was performed on a ${\mu}-Bondapak$ $C_{18}$ column with a solvent system of acetonitrile : dichloromethane : methanol = 70:20:10. To check the reliability of the method applied, precision and recovery tests were performed. Perilla leaf showed the highest ${\beta}-carotene$ content(12,570 ${\mu}g$ / 100 g), followed by mugwort and chwi, all of those have ${\le}10,000\{\mu}g\{\beta}-carotene$ per 100 g vegetables. Green lettuce, spinach, leek, crown daisy and reddish brown lettuce contained 9,869, 6,689, 5,664, 3,601 and 3,299 ${\mu}g\{\beta}-carotene/100 g$, respectively, Lutein content was the highest in perilla leaf($13,718{\mu}g/100 g$) followed by chwi($11,989{\mu}g/100 g$), mugwort($11,522{\mu}g/100 g$), green lettuce($10,307{\mu}g/100 g$) and spinach($10,115{\mu}g/100 g$). ${\beta}-carotene$ contents in perilla leaf, mugwort, chwi and green lettuce were 47.8~49.6% of total carotenoids, and ${\beta}-carotene$ contents in the other green leafy vegetables analyzed were 37.7~41.4% Vitamin A contents of green leafy vegetables analyzed by HPLC were 2~6 times higher than the vitamin A values shown in food composition tables except crown daisy.

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Development of Content Analysis for Cyperus rotundus by HPLC-UV and a Comparison between Chinese and Domestic Cyperi Rhizoma (HPLC-UV 방법을 이용한 향부자 분석법 개발과 국산 및 중국산 향부자의 패턴비교)

  • Seo, Jiyun;Kim, Jinah;Kim, Sungun;Liu, Ting;Whang, Wan Kyunn
    • YAKHAK HOEJI
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    • v.56 no.5
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    • pp.280-287
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    • 2012
  • A high-performance liquid chromatography (HPLC) combined with ultraviolet (UV) method for the simultaneous determination of ${\alpha}$-cyperone and nootkatone was developed for the quality control of Cyperus rotundus Linne. The separation was performed on a KR100-$5C_{18}$ ($4.6{\times}250mm$) column, and an elution gradient composed of methanol and water with a flow-rate of 1.0 ml/min. Detection wavelength was set at 254 nm. The optimum extraction for the detection of the ${\alpha}$-cyperone and nookatone was achieved by ultrasonic with methanol for an hour. Two marker compounds ${\alpha}$-cyperone and nootkatone in Cyperi Rhizoma showed good linearity ($R^2$ >0.999) in the concentration range of $12.5{\mu}g/ml$ to $200{\mu}g/ml$. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.04~1.23% and 0.08~0.68%, respectively, and the overall recoveries of 97.45~105.58% for the two compounds analyzed. Additionally, a difference was observed in the cluster analysis and principal component analysis between Cyperi Rhizoma in Korea and China. The result demonstrated that the principal component analysis is useful to distinguish between Cyperi Rhizoma in Korea and China.

Purification and Characterization of Transglutaminase from a Newly Isolated Streptomyces platensis YK-2 (토양 방선균 Streptomyces platensis YK-2가 생산하는 Transglutaminase의 정제 및 효소학적 특성)

  • Ko, Hee-Sun;Kim, Hyun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.6
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    • pp.801-806
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    • 2009
  • A species producing transglutaminase (EC 2.3.2.13) was isolated from forest soil and identified as Streptomyces platensis YK-2. The transglutaminase was purified from culture broth by 50% methanol precipitation, followed by successive chromatography on DEAE-Sephadex. The yield and purification-fold was 63.4% and 2.2-fold, respectively. The purified microbial transglutaminase (MTG) migrated as a single band of approximately 45 kDa upon sodium dodecyl sulfate polyacrylamide gel eletrophoresis. The isoelectric point determined by multichambered electrofocusing was pH $6.0{\sim}7.0$. The enzyme was strongly inhibited by $Hg^{++}$, but was activated by $Cd^{++}$, $Mg^{++}$, $Mn^{++}$, $Pb^{++}$ and reducing agents such as dithiothreitol and mercaptoethanol.

Purification and Characterization of Beta-Glucosidase from Weissella cibaria 37

  • Lee, Kang Wook;Han, Nam Soo;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1705-1713
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    • 2012
  • A gene encoding ${\beta}$-glucosidase was cloned from Weissella cibaria 37, an isolate from human feces. Sequence analysis showed that the gene could encode a protein of 415 amino acids in length, and the translated amino acid sequence showed homology (34-31%) with glycosyl hydrolase family 1 ${\beta}$-glucosidases. The gene was overexpressed in E. coli BL21(DE3) using pET26b(+) and a 50 kDa protein was overproduced, which matched well with the calculated size of the enzyme, 49,950.87 Da. Recombinant ${\beta}$-glucosidase was purified by using a his-tag affinity column. The purified ${\beta}$-glucosidase had an optimum pH and a temperature of 5.5 and $45^{\circ}C$, respectively. Among the metal ions (5mM concentration), $Ca^{2+}$ slightly increased the activity (108.2%) whereas $Cu^{2+}$ (46.1%) and $Zn^{2+}$ (56.7%) reduced the activity. Among the enzyme inhibitors (1 mM concentration), SDS was the strongest inhibitor (16.9%), followed by pepstatin A (45.2%). The $K_m$ and $V_{max}$ values of purified enzyme were 4.04 mM and 0.92 ${\mu}mol/min$, respectively, when assayed using pNPG (p-nitrophenyl-${\beta}$-D-glucopyranoside) as the substrate. The enzyme liberated reducing sugars from carboxymethyl cellulose (CMC).

Chemical Standardization of Poria cocos (복령의 품질표준화를 위한 지표성분 탐색 및 정량법 개발)

  • Hoang, Lam;Kwon, Soon-Ho;Kim, Kyung-Ah;Hur, Jong-Moon;Kang, Young-Hwa;Song, Kyung-Sik
    • Korean Journal of Pharmacognosy
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    • v.36 no.3 s.142
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    • pp.177-185
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    • 2005
  • For the purpose of the quality control of Poria cocos, three major compounds were isolated and identified as pachymic acid (1), $3{\beta}-hydroxylanosta-7,9(11),24-trien-21-oic$ acid (2), and dehydroeburicoic acid (3). The optimal extraction conditions for the quantification of pachymic acid its analogues were the 3 hours of reflux with 15g of P. cocos in 100ml 95% ethanol. HPLC conditions were as follows: Column; ZORBAX Eclipse XDB C18 $(4.6{\times}250\;mm,\;Agillent)$, mobile phase; 1% HOAc in 70% $MeOH{\rightarrow}1%$ HOAc in 100% MeOH for 25 min, then 1% HOAc in 100% MeOH for 15 min, detector, ELSD, flow rate; 1ml/min. The mean contents of 1,2, and 3 in Poria cocos cultivated at 18 different site were $0.65{\pm}0.19,\;0.88{\pm}0.72,\;and\;0.84{\pm}0.54\;mg/g$, respectively, and values might be the guide line for the quality control of P. cocos.