• Title/Summary/Keyword: HPLC screening

Search Result 159, Processing Time 0.024 seconds

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
    • /
    • v.34 no.3
    • /
    • pp.505-509
    • /
    • 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).

Screening of Domoic Acid, a Marine Neurotoxin, in Korean Shellfishes (국내산 패류의 신경독소 domoic acid 검색)

  • Koh, Eun-Mi;Kwon, Hoon-Jeong
    • Korean Journal of Food Science and Technology
    • /
    • v.34 no.6
    • /
    • pp.1130-1133
    • /
    • 2002
  • Domoic acid, and amnesic shellfish poison, is a neurotoxin frequently found in shellfishes. Guidance level for the consumable shellfish has been established as $20\;{\mu}g$ domoic acid/g by Health and Welfare Canada and U.S. FDA. Domoic acid is produced by pennate diatom, a Nitzschia pungens f. multiseries ingested by the shellfish. Content of domoic acid in shellfish samples collected along the Korean shoreline from May to December of 1999 was analyzed. The collection included 1 Gastropoda (Murex shell) and 11 Bivalvias (oyster, little neck clam, orient hard clam, venus clam, surf clam, ark shell, hard-shelled mussel, pen shell, jack-knife clam, pink butterfly shell, and granulated ark shell). Samples were homogenized, extracted with 50% methanol, filtered, and analyzed by reversed-phase liquid chromatography at 242 nm with mobile phase consisting of 10% acetonitrile and 0.1% trifluoroacetic acid. Recovery of the HPLC analysis was 95.80% (${\pm}1.09$). All tested samples showed no domoic acid at the detection limit of 50 ng/g.

Isolation and Characterization of Helicobacter pylori Urease Inhibitor from Rubus coreanus Miquel (복분자(Rubus coreanus Miquel)로부터 Helicobacter pylori Urease Inhibitor의 분리 및 특성)

  • 양성우;호진녕;이유현;신동훈;홍범식;조홍연
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.33 no.5
    • /
    • pp.769-777
    • /
    • 2004
  • A Helicobacter pylori urease inhibitor from Rubus coreanus Miquel has been isolated and partially characterized for aiming to Prevent H. pylori growth and decrease harmful accumulation of ammonia in human gastric mucosa. We screened urease inhibitory activities in 519 extracts library prepared by solvent extraction from 173 kinds of edible plants, medicinal herbs, herbs and seaweeds using a colorimetric urease assay system. As results of primary and secondary screening, 70% acetone extract of Rubus coreanus Miquel was selected as potent candidate, showing about 24% inhibitory activity. The acetone extract was sequentially partitioned into RCE/RCWI and RCB/RCW2 layers with ethyl acetate and butanol. The major active component in RCW2, water layer from butanol fractionation was revealed to be peptidic or proteinous substance by inhibitory activity determination after pronase digestion and periodate oxidation. RCW2-IIIc a was isolated by sequential column chromatography on DEAE-Toyopearl 650C, Butrl-Toyopearl 650M and Sephadex LH-20. The isolated urease inhibitor RCW2-IIIc $\alpha$, was highly pure proteinous substance with molecular weight of 13kDa by high-performance gel permeation liquid chromatography. RCW2-IIIc$\alpha$ has about 5 times higher inhibitory activity than 70% acetone extract, showing high stability against heat treatment and peptic digestion.

Pesticides Residue Monitoring and Impact Evaluation of Golf Course and Neighbouring Area in Korea (국내 골프장농약 사용에 따른 골프장 및 인근 지역의 잔류농약 모니터링 조사를 통한 영향평가)

  • Lee, Jun-Bae;Cho, Hoon-Je;Kwak, Eun-Jie;Park, Kyoung-Hoon;Lee, Min-Keong;Kim, Hyun-Koo;Jeoung, Hyeon-Mi;Chang, Hee-Ra
    • Korean Journal of Environmental Agriculture
    • /
    • v.37 no.4
    • /
    • pp.277-282
    • /
    • 2018
  • BACKGROUND: At these days, the human health and environmental concerns of pesticide used for turf grass management at golf courses in Korea have increased. The objectives of the study were to determine the pesticide residues for golf course and neighboring area and evaluate the impact moved into neighboring area of pesticides treated at golf courses. METHODS AND RESULTS: Three golf courses and neighboring areas in Korea were monitored from July to October, 2017. The soil sample collection was divided the golf course into its logical parts (such as a greens, fairways, and rough) and neighboring area soil samples were collected at three different points. The water samples of the golf course and neighboring area were collected at three different points, respectively. The pesticide residues for soil and water sample were monitored by the multi-residue screening method of 98 pesticide with HPLC-MS-MS. The concentrations of detected pesticide in soil and water samples of the golf course were in the range of 0.01~1.26 mg/kg and 0.0001~0.0089 mg/kg, respectively. The residue levels for detected pesticides in neighboring area were at 0.01~0.04 mg/kg and 0.0001~0.0029 mg/kg, respectively, well below those level in golf course. CONCLUSION: This study indicate that the pesticide residue levels of golf course and neighboring area in Korea may not a possible risk of exposure on soil and aquatic environment. For future work, more monitoring should be performed so that the evaluation data becomes more valid.

Establishment of a Nondestructive Analysis Method for Lignan Content in Sesame using Near Infrared Reflectance Spectroscopy (근적외선분광(NIRS)을 이용한 참깨의 lignan 함량 비파괴 분석 방법 확립)

  • Lee, Jeongeun;Kim, Sung-Up;Lee, Myoung-Hee;Kim, Jung-In;Oh, Eun-Young;Kim, Sang-Woo;Kim, MinYoung;Park, Jae-Eun;Cho, Kwang-Soo;Oh, Ki-Won
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.67 no.1
    • /
    • pp.61-66
    • /
    • 2022
  • Sesamin and sesamolin are major lignan components with a wide range of potential biological activities of sesame seeds. Near infrared reflectance spectroscopy (NIRS) is a rapid and non-destructive analysis method widely used for the quantitative determination of major components in many agricultural products. This study was conducted to develop a screening method to determine the lignan contents for sesame breeding. Sesamin and sesamolin contents of 482 sesame samples ranged from 0.03-14.40 mg/g and 0.10-3.79 mg/g with an average of 4.93 mg/g and 1.74 mg/g, respectively. Each sample was scanned using NIRS and calculated for the calibration and validation equations. The optimal performance calibration model was obtained from the original spectra using partial least squares (PLS). The coefficient of determination in calibration (R2) and standard error of calibration (SEC) were 0.963 and 0.861 for sesamin and 0.875 and 0.292 for sesamolin, respectively. Cross-validation results of the NIRS equation showed an R2 of 0.889 in the prediction for sesamin and 0.781 for sesamolin and a standard error of cross-validation (SECV) of 1.163 for sesamin and 0.417 for sesamolin. The results showed that the NIRS equation for sesamin and sesamolin could be effective in selecting high lignan sesame lines in early generations of sesame breeding.

A Simple Method for Evaluation of Pepper Powder Color Using Vis/NIR Hyperspectral System (Vis/NIR 초분광 분석을 이용한 고춧가루 색도 간이 측정법 개발)

  • Han, Koeun;Lee, Hoonsoo;Kang, Jin-Ho;Choi, Eunah;Oh, Se-Jeong;Lee, Yong-Jik;Cho, Byoung-Kwan;Kang, Byoung-Cheorl
    • Horticultural Science & Technology
    • /
    • v.33 no.3
    • /
    • pp.403-408
    • /
    • 2015
  • Color is one of the quality determining factors for pepper powder. To measure the color of pepper powder, several methods including high-performance liquid chromatography (HPLC), thin layer chromatography (TLC), and ASTA-20 have been used. Among the methods, the ASTA-20 method is most widely used for color measurement of a large number of samples because of its simplicity and accuracy. However it requires time consuming preprocessing steps and generates chemical waste containing acetone. As an alternative, we developed a fast and simple method based on a visible/near infrared (Vis/NIR) hyperspectral method to measure the color of pepper powder. To evaluate correlation between the ASTA-20 and the visible/near infrared (Vis/NIR) hyperspectral methods, we first measured the color of a total of 488 pepper powder samples using the two methods. Then, a partial least squares (PLS) model was postulated using the color values of randomly selected 3 66 samples to predict ASTA values of unknown samples. When the ASTA values predicted by the PLS model were compared with those of the ASTA-20 method for 122 samples not used for model development, there was very high correlation between two methods ($R^2=0.88$) demonstrating reliability of Vis/NIR hyperspectral method. We believe that this simple and fast method is suitable for highthroughput screening of a large number of samples because this method does not require preprocessing steps required for the ASTA-20 method, and takes less than 30 min to measure the color of pepper powder.

Studies on the Function of the Rv2435c Gene of the Mycobacterium bovis BCG (Mycobacterium bovis BCG Rv2435c 유전자의 기능에 대한 연구)

  • Lee Seung-Sil;Bae Young-Min
    • Journal of Life Science
    • /
    • v.15 no.3 s.70
    • /
    • pp.415-422
    • /
    • 2005
  • library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

Screening of Lignan Compounds and Antioxidant Activity of Chungkukjang Fermented with Defatted Sesame Flour (참깨 탈지박을 첨가하여 제조한 청국장의 생리활성 및 리그난 성분 탐색)

  • Kim, Tae-Su;Choi, Mi-Kyeong;Kim, Jin-Sook;Han, Jae-Woong;Kang, Myung-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.38 no.11
    • /
    • pp.1580-1586
    • /
    • 2009
  • This study is on the effect of oil seed by-products added to Chungkukjang. For this, we designed three cases: Chungkukjang was added in with defatted sesame flour before fermented (DSFBF), added with defatted sesame flour after fermented (DSFAF) and with no adding (control). In each case, the common ingredients and the active antioxidant ingredients were examined and compared and the effects were analyzed. According to microanalysis result, carbohydrate content Chungkukjang the DSFAF 24.97%, control 23.86%, DSFBF Chungkukjang 20.21% as compared to control and Chungkukjang DSFAF relatively low carbohydrate content. The moisture contents in DSFBF (55.98%) or DSFAF (52.83%) were higher than that in control (48.89%). Chungkukjang crude ashes in DSFBF (1.48%) or DSFAF (2.41%) were much lower than in control (6.45%). The proportions of crude lipid in DSFBF (3.30%) or DSFAF (3.93%) were higher than in control (1.77%) by about 2%. As for crude protein, the percentage in DSFAF (15.86%) was lower than that of DSFBF (19.03%) or of control (19.03%). There was no meaningful difference in biological activity measurement as total phenolic contents were 1.26 mg/mL in DSFBF, 1.14 mg/mL in DSFAF and 1.26 mg/mL in control. But electron donating ability was meaningfully more active in DSFBF (21.30%) than in control (20.24%). The superoxide dismutase (SOD)-like activity in DSFBF (68.48%) was twice higher than in control (34.01%), which may imply that DSFBF contain some ingredients that can scavenge superoxide anion radically. In hydroxyl radical scavenging activity, DSFAF scores 96.87%, which is the highest with 96.40% in DSFBF and 95.73% in control. Relative antioxidative effects in DSFBF was 47.92%, which is comparable to 47.06% in control. As a result of extraction and quantitative HPLC analysis of sesamin and sesamolin extracted from the samples, DSFBF contained 3.04$\pm$0.21 mg/g of sesamin, which is meaningfully higher than 2.41$\pm$0.14 mg/g in DSFAF. Content of sesamolin was higher in DSFBF (1.36$\pm$0.09 mg/g) than DSFAF (1.12$\pm$0.07 mg/g) or in control. We can conclude that biologically active and effective ingredients could be found more in DSFBF than in DSFAF or in control. This study conveys not only the meaning that oil seed by-products can be used as an ingredient for making Chungkukjang functional food, but also the possibility that oil seed by-products themselves could become excellent functional food.

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
    • /
    • v.35 no.4 s.139
    • /
    • pp.368-374
    • /
    • 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.