• Title/Summary/Keyword: RP HPLC

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The VHR Dual-Specificity Protein Tyrosine Phosphatase (DS-PTPase) Inhibitor Isolated from Gastrodia elata (천마(Gastrodia elata)로부터 분리한 VHR DS-PTPase 저해 물질)

  • Lee, Myung-Sun;Oh, Won-Keun;Bae, Eun-Young;Ahn, Soon-Cheol;Sohn, Cheon-Bae;Osada, Hiroyuki;Ahn, Jong-Seog
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.505-509
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    • 2002
  • In the course of screening program for VHR DS-PTPase (dual-specificity protein tyrosine phosphatase) from natural sources, Gastrodia elata was selected. One compound showing potent inhibitory activity was isolated by the solvent extraction and column chromatography including silica gel, ODS RP-18, Sephades LH-20, and HPLC. This compound was identified as baicalein by several NMR techniques such as $^1H-NMR$, $^{13}C-NMR$, and DEPT. Baicalein showed selective inhibitory activity against VHR DS-PTPase with $IC_{50}=2.4\;{\mu}M$, and showed cytotoxicity against several human cancer cell lines with an $GI_{50}$ of $5.26{\sim}12.93\;{\mu}g/mL$ range, including, melanoma (LOX-IMVI), lung cancer (NCI H23 and A549), colon cancer (HCT 116 and SW 620), prostate cancer (PC-3), and leukemia (MOLT 4F).

Antimicrobial effect of Enterococcus faecalis BMSE-HMP005 isolated from human breast milk against multidrug-resistant bacteria (모유 유래 유산균 Enterococcus faecalis BMSE-HMP005의 다제내성 균에 대한 항균효과)

  • Lee, Jeong-Eun;Kim, Soo-bin;Yu, Du-na;Jo, So-Yeon;Kim, Ae-Jung;Kook, Moochang
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.209-217
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    • 2022
  • In this study, Enterococcus faecalis BMSE-HMP005 isolated from human breast milk demonstrated antimicrobial effects against multidrug-resistant (MDR) bacterial strains. The bacteriocin produced by E. faecalis BMSE-HMP005 was fractionated using reverse-phase high-performance liquid chromatography. This fraction showed antimicrobial effects against both gram-positive and gram-negative MDR bacteria. No hemolytic reactions were observed. E. faecalis BMSEHMP005 was resistant to vancomycin; however, kanamycin, ampicillin, and erythromycin showed minimum inhibitory concentrations that were lower than the acceptable range provided by the European Food Safety Authority. For artificial gastric juice and bile acid, the survival rates were 98.67% and 95.70%, respectively. These results show the potential utility of E. faecalis BMSE-HMP005 as a probiotic with remarkable antimicrobial effects against MDR bacteria.

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.

Development and Physical Properties of Low-Trans Spread Fat from Canola and Fully Hydrogenated Soybean Oil by Lipase-Catalyzed Synthesis (카놀라유와 대두극도경화유로부터 효소적으로 합성된 저트랜스 스프레드 고체지의 특성)

  • Kim, Young-Joo;Lyu, Hyun-Kyeong;Shin, Jung-Ah;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.9
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    • pp.1328-1334
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    • 2010
  • Low-trans spread fat (LTSF) was produced by lipase-catalyzed synthesis of canola (CO) and fully hydrogenated soybean oil (FHSBO) at 65:35 (w/w). Blend of CO and FHSBO with 65:35 ratio was interesterified using Lipozyme TLIM (immobilized Thermomyces lanuginosus, 20% of total substrate) in a 1 L-batch type reactor at $70^{\circ}C$ with 500 rpm for 24 hr. Then, physicochemical melting properties of LTSF were compared with commercial spread fat. At $20^{\circ}C$, solid fat contents (SFC) of commercial spread fat as a control and LTSF were similar, showing 19.1 and 18.1%, respectively. Major compositional fatty acids of LTSF were C18:0, C18:1 and C18:2 (29.2, 41.8 and 13.3 wt%, respectively). Trans fatty acid content of the LTSF (0.2 wt%) was lower than that of commercial spread fat (5.5 wt%). In the RP-HPLC analysis from LTSF, major triacylglycerol (TAG) molecules were SOL (stearoyl-oleoyl-linoleyl), SOO, POS/PSP, and SOS. Also, polymorphic form and x-ray diffraction of LTSF showed coexistence of $\beta$' and $\beta$ form crystals.

Studies on Simultaneous Determination of Chlorophyll a and b, Pheophorbide a, and $\beta-Carotene$ in Chlorella and Spirulina Products (클로렐라 및 스피루리나제품에 함유된 엽록소 a, b, 페오포르바이드 a 및 $\beta$-카로틴의 동시분석법에 관한 연구)

  • Lee Young Ja;Kim So Hee;Kim Jin-Sook;Han Jeong A;Seo Hae Jeom;Lim Hyo Jeong;Choi Soo Young
    • Journal of Food Hygiene and Safety
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    • v.20 no.3
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    • pp.141-146
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    • 2005
  • A simple and sensitive analysis method based on reverse phase (RP) HPLC with UV detector was developed for simultaneous determination of chlorophyll a and b, pheophorbide a and $\beta-Carotene$ in Chlorella and Spirulina products. For added concentration $(50\;\mug/ml)$ of chlorophyll a and b, pheophorbide a and $\beta-Carotene$, recoveries of those were 70.3, 71.6, 60.1 and $90.5\%$, respectively, with relative standard deviations of 2.8,6.0, 10.6 and $10.4\%$. Limit of detection and quantification had ranges of $0.1\sim1.0\;\mug/ml$ and $0.2\sim2.0\;\mug/ml$, respectively. Calibration curve was linear with correlation coefficient of 0.995 for chlorophyll a and b, pheophorbide a and $\beta-Carotene$. Results of simultaneous determination in Chlorella and Spirulina products were showed ranges of $121.g\sim543.0\;\mug/ml$ for chlorophyll a,$0.6\sim160.0\;\mug/ml$ for chlorophyll b, $19.2\sim60.3\;\mug/ml$ for pheophorbide a and $383.6\sim1713.7\;\mug/ml$ for $\beta-Carotene$, respectively. Chlorophyll b contents in Chlorella products were detected above 30 times level to those in Spirulina products. $\beta-Carotene$ contents in Spirulina products were detected 2.7 times level to those in Chlorella products.

Comparision of Preparation Methods for Water Soluble Vitamin Analysis in Foods by Reversed-Phase High Performance Liquid Chromatography (역상 고속 액체 크로마토그래피에 의한 식품 중 수용성 비타민 분석을 위한 전처리법의 비교)

  • Kim, Hyung-Soo;Jang, Duck-Kyu;Woo, Dong-Kyun;Woo, Kang-Lyung
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.141-150
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    • 2002
  • Owing to a need for simple extraction and purification for analysis of water soluble vitamins in food samples by RP-HPLC with UV-detector, the methods of bromelain and protease hydrolysis and $C_{18}$ Sep-Pak solid phase extraction were employed. The recoveries of standard water soluble vitamins by the bromelain and protease hydrolysis and $C_{18}$ Sep-Pak solid phase extraction were significantly high compared to AOAC methods in most of vitamins. The contents of pyridoxal determined with protest in the pork was similar, but in the bromelain hydrolysis and AOAC method, was high compared to the results of reference. The niacinamide, thiamin and riboflavin determined with bromelain and protease hydrolysis showed similar values to the results of references. In the potato, pyridoxamine was detected in the AOAC method, which was not detected in the bromelain and protease hydrolysis methods. Pyridoxal contents in the protease hydrolysis and AOAC methods were very similar to the results of references. The recoveries of fortified standard vitamins in food samples were significantly high and accurate compared to those of AOAC methods. The extraction and purification with $C_{18}$ Sep-Pak solid extractor might be considered superior method for the determination of water soluble vitamins in food samples.

Determination of S-Allyl-L-cystein, Diallyl Disulfide, and Total Amino Acids of Black Garlic after Spontaneous Short-term Fermentation (자가숙성발효 후 흑마늘의 S-Allyl-L-cystein, Diallyl Disulfide 및 Total Amino Acids 분석)

  • Kim, Mun-Su;Kim, Min-Ju;Bang, Woo-Suk;Kim, Keun-Sung;Park, Sung-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.5
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    • pp.661-665
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    • 2012
  • Garlic (Allium sativum L.) is one of the oldest cultivated plants and has been used throughout the world as a food supplement and a folk medicine for thousands of years. Raw garlic has been processed into a variety of commercial garlic products for consumer convenience. The latest new processing technology, 'spontaneous short-term fermentation', has been developed to process raw garlic into black garlic. The physiologically active effects of garlic have been attributed to its organosulfur compounds. In this study, the proximate compositions and the total amino acid content of raw Namhae garlic and black garlic were determined. The two major organosulfur compounds of garlic, $S$-allyl-L-cysteine (SAC), and diallyl-disulfide (DADS), were also analyzed using RP-HPLC. The proximate compositions were not different between raw and black garlic. The amount of 13 amino acids was greater in black garlic than in raw garlic among a total of 17 amino acids considered. The black garlic had 2-fold higher levels of SAC and 30-fold higher levels of DADS than the raw garlic. Therefore, it is suggested that consuming black garlic produced by spontaneous short-term fermentation is more effective than consuming raw garlic, in order for consumers to take more physiologically active organosulfur compounds (SAC and DADS), which are the compounds that are good for consumer health.

Effects of 3,5-di-O-Caffeoylquinic acid from Artemisia scoparia Waldstein et Kitamura on the Function of HPV 16 Oncoproteins (인진에서 분리한 3,5-di-O-Caffeoylquinic acid가 자궁경부암 바이러스 발암단백질의 기능에 미치는 영향)

  • Baek, Tae-Woong;Lee, Kyung-Ae;Ahn, Min-Jung;Joo, Hae-Kyung;Cho, Min-Chul;Kang, Jung-Woo;Kim, Hee-Seo;Shim, Jung-Hyun;Lee, Hee-Gu;Oh, Hyun-Cheol;Ahn, Jong-Seok;Cho, Yong-Kwen;Myung, Pyung-Keun;Yoon, Do-Young
    • Korean Journal of Pharmacognosy
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    • v.35 no.4 s.139
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    • pp.368-374
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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 papillomavirus) type 16. The HPV is 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, Artemisia scoparia Waldstein et Kitamura has inhibitory effects on the binding between E6 oncoprotein and tumor suppressor p53, or the binding between E6 and E6 associated protein (E6AP), an E3 ubiquitin-protein ligase. HPV oncoprotein inhibitors from Artemisia scoparia W. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and the inhibitory compounds were finally purified by HPLC using an ELISA screening system based on the 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 the functions of E6 oncoprotein. We investigated whether 3,5-di-O-caffeoylquinic acid (DCQA) extracted from Artemisia scoparia W. Could inhibit the function of E6 oncoprutein. DCQA 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 DCQA 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.

Immobilization of Burkholderia cepacia Lipase on Weak Base Styrene Resin Using Polyethyleneimine with Cross-linking (PEI(Polyethyleneimine)를 이용하여 음이온계 레진에 고정화된 Lipase AH 제조 및 효소적 Interesterification을 통한 반응 특성 연구)

  • Lee, Chi Woo;Lee, Ki Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.7
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    • pp.1025-1035
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    • 2014
  • This study assessed the effect of immobilized lipase on weak base styrene resin using polyethyleneimine (PEI) with cross-linking. Two procedures were used in this study. The first one, "mono-layer" lipase immobilization, involves washing PEI after adsorption. The second procedure, "multi-layer" lipase immobilization, has no washing before the cross-linking step. Treverlite XS-100200 (weak base styrene resin) was immersed with PEI solution (2.2 mg/mL). Lipase AH (from Burkholderia cepacia) was adsorbed onto the support coated with PEI before cross-linking with glutaraldehyde. Structured lipid was synthesized by immobilized lipase-catalyzed interesterification using canola oil, palmitic ethyl ester (PEE), and stearic ethyl ester (StEE). Total fatty acid contents of triacylglycerol (TAG) in structured lipids were analyzed to investigate activity, properties, and reusability of immobilized lipases. Activities of immobilized lipases on the multi-layer and mono-layer increased at a high concentration (8 mg/mL) of lipase solution used for immobilization. The results show that immobilized lipase with the mono-layer method at pH 8.0 on resin had the highest total saturated fatty acid content (26.17 area%). Activity of immobilized lipase with the multi-layer method at pH 7.5 on support was lower than that of the mono-layer, but total saturated fatty acid content was 16.79 area% higher than that of lipase AH (15.01 area%).

Antioxidant Activity of Different Parts of Lespedeza bicolor and Isolation of Antioxidant Compound (싸리나무(Lespedeza bicolor) 부위별 추출물의 항산화 활성 및 항산화물질 분리)

  • Lee, Jae-Hak;Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.763-771
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    • 2012
  • In this study, total antioxidant properties of extracts from different parts of Lespedeza bicolor were determined using techniques of measuring 1,1-diphenyl-2-picryl hydrazyl/2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)-radical scavenging activity and total phenolic contents. The total antioxidant activities of leaf, stem and root extracts from various solvents (water, 50, 70, 100% ethanol, and hot-water) indicated that 50 and 70% ethanol extracts have high radical scavenging activities and phenolic contents. A systematic approach was used to determine the total antioxidant activity of different solvent fractions of the Lespedeza bicolor extracts, partitioning with chloroform, ethyl acetate, n-butanol, and water, and the ethyl acetate fraction was found to have the strongest antioxidant activity. Antioxidant assay-guided isolation was carried out to isolate potential antioxidant compounds. The ethyl acetate fraction of the leaf extract was subjected to silica gel, LH-20 and RP-18 column chromatography successively, and afforded compound 1, which was identified as eriodictyol by NMR and MS analysis, after which its antioxidant activity was determined.