• Title/Summary/Keyword: RP HPLC

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Free Amino Acids and Their Derivatives of Ganoderma lucidum (영지 버섯의 유리 아미노산 및 유도체에 관한 연구)

  • 이현아;김병각;현진원
    • Environmental Mutagens and Carcinogens
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    • v.24 no.2
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    • pp.67-72
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    • 2004
  • To find active components in Ganoderma lucidum, their free amino acid and free amino acid derivatives were analyzed. After extracting with hot water, the extracts were filtrated by three steps. So, supernatants below 10,000 dalton were obtained. Filtrates were derivativated with PITC (phenylthiocarbamyl) derivative reagent and PITC amino acid was obtained. Then, they were analyzed by RP-HPLC. 13 amino acids were analyzed in cultured Korean Nok-kak ji (one of Ganoderma lucidum), 15 amino acids in cultured Korean Ganoderma lucidum, 12 amino acids wild Ganoderma lucidum, 16 amino acids in cultured Taiwan Ganoderma lucidum.

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Method Development of Verapamil in Presence of NSAIDs using RP-HPLC Technique

  • Sultana, Najma;Arayne, M. Saeed;Waheed, Abdul
    • Bulletin of the Korean Chemical Society
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    • v.32 no.7
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    • pp.2274-2278
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    • 2011
  • Verapamil is a calcium channel blocker and is classified as a class IV anti-arrhythmic agent. It is used in the control of supra ventricular tachyarrhythmias, and in the management of classical and variant angina pectoris. It is also used in the treatment of hypertension and used as an important therapeutic agent for angina pectoris, ischemic heart disease, hypertension and hypertrophic cardiomyopathy. Verapamil commonly co-administered with NSAIDs (non-steroidal anti-inflammatory drugs) i.e. diclofenac sodium, flurbiprofen, Ibuprofen, mefanamic acid and meloxicam. A simple and rapid RP-HPLC method for simultaneous determination and quantification of verapamil and NSAIDs was developed and validated. The mobile phase constituted of acetonitrile: water (55:45) whose pH was adjusted at 2.7 and pumped at a flow rate of 2.0 mL $min^{-1}$ at 230 nm. The proposed method is simple, precise, accurate, low cost and least time consuming for the simultaneous determination of verapamil and NSAIDs which can be effectively applied for the analysis of human serum.

Efficient Isolation of Dihydrophaseic acid 3'-O-β-D-Glucopyranoside from Nelumbo nucifera Seeds Using High-performance Countercurrent Chromatography and Reverse-phased High-performance Liquid Chromatography

  • Rho, Taewoong;Yoon, Kee Dong
    • Natural Product Sciences
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    • v.24 no.4
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    • pp.288-292
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    • 2018
  • High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase highperformance liquid chromatography (RP-HPLC) method was developed to isolate dihydrophaseic acid 3'-O-${\beta}$-D-glucopyranoside (DHPAG) from the extract of Nelumbo nucifera seeds. Enriched DHPAG sample (2.3 g) was separated by HPCCC using ethyl acetate/n-butanol/water system (6:4:10, v/v/v, normal-phase mode, flow rate: 4.0 mL/min) to give 23.1 mg of DHPAG with purity of 88.7%. Further preparative RP-HPLC experiment gave pure DHPAG (16.3 mg, purity > 98%). The current study demonstrates that utilization of CCC method maximizes the isolation efficiency compared with that of solid-based conventional column chromatography.

Simultaneous Analysis of Both Lactone Form and Acid Form Monacolin K in Red Yeast Rice by RP-HPLC (역상 HPLC에 의한 홍국 중의 락톤 및 산성 모나콜린 K의 동시분석법)

  • Moon, Young-ja;Wang, Qi-jun;Xu, Bao-jun;Li, Chang-tian;Kim, Jae-hoon;Mo, En-kyeng;Baek, Seoung-young;Il Kwon;Sung, Chang-keun
    • The Korean Journal of Food And Nutrition
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    • v.14 no.6
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    • pp.521-526
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    • 2001
  • A method for the simultaneous and precise determination of lactone form and acid form monacolin K in red yeast rice by HPLC was developed in this study. The standard of acid form monacolin K was prepared by alkaline hydrolysis of its lactone form, which was purchased from Sigma company. The optimum HPLC system for the separation and quantification of acid form and lactone form monacolin K is based on the reversed-phase column, and the acidified mobile phase consisting of acetonitrile : 0.1% trifluoroacetic acid (TFA) water soln : 62 :38, the low limit detection amount was 5 ng (i.e.10 $\mu$l injection of 0.5 $\mu\textrm{g}$/ml) . And the optimal extracting system for monacolins in red rice was also presented here.

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Purification and Characterization of an Insect Antibacterial Peptide, Defensin, Expressed in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 발현한 곤충 항균펩티드, defensin의 정제 및 특성 조사)

  • 강대욱;이준원;김보연;안종석
    • Journal of Life Science
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    • v.12 no.4
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    • pp.483-489
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    • 2002
  • We investigated the biochemical properties of insect defensin expressed and secreted from Saccharomyces corevisiae. The defensin showed extremely high resistance to boiling for up to 30 min and to pH values tested from 2.0 to 12.0. The treatment of defensin with various proteases abolished antibacterial activity. However, amylases, cellulase, lipase and catalase had no effect on the activity. The defensin was purified to homogeneity through ammonium sulfate concentration of culture supernatant, SP-Sepharose column chromatography and RP-HPLC. Tricin-SDS-PAGE analysis revealed that the molecular weight of the defensin was about 4.0 kDa. The antibacterial activity of the purified defensin was verified by renaturation of stained gel and gel pouring assay using Micrococcus luteus as a test organism.

Analysis of Gingerol Compounds of Raw Ginger (Zingiber officinale Roscoe) and Its Paste by High Performance Liquid Chromatograph-Mass Spectrometry (LC/MS) (LC/MS에 의한 원료생강 및 생강 페이스트 중의 Gingerol 화합물 분석)

  • 조길석
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.5
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    • pp.747-751
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    • 2000
  • This study was carried out to improve the analysis method of gingerol compounds from ginger (Zingiber officinale Roscoe). Pungent components of ginger were extracted by acetone and lisolated by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) with LiChrosorb RP-18 column. Three homologues of gingerols were identified by HPLC-mass spectrometry (LC/MS) and nuclear magnetic resonance (NMR). The contents of [6]-, [8]- and [10]-gingerols in three homologues identified were 635.3 mg%, 206.6 mg% and 145.7 mg% in raw ginger, and were 418.2 mg%, 142.6 mg% and 103.3 mg% in ginger paste, respectively.

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Isolation and Partial Characterization of Phytotoxic Mycotoxins Produced by Sclerotinia sp., a Potential Bioherbicide for the Control of White Clover(Trifoliorum repens)

  • Hong, Yeon-Kyu;Lee, Bong-Choon;Jung, Won-Kwon;Bae, Soon-Do;Park, Sung-Tae;Uhm, Jae-Youl
    • The Plant Pathology Journal
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    • v.20 no.1
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    • pp.52-57
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    • 2004
  • Sclerotinia sp. (isolate BWC98-105) causes stem blight and root rot in Leghum sp., and is presently being evaluated as a potential mycoherbicide for the control of Trifoliorium repens. Bioassays have shown that Sclerotinia sp. produces phytotoxic substance which is biologically active against T. repens. Two biologically active compounds, designated as compoundsI and II, were produced in vitro from the culture filtrate of BWC98-105 isolate Sclerotium sp. Compounds I and II were purified by means of liquid-liquid extraction and $C_{18}$ open column chromatography (300 ${\times}$ 30 mm, i.d). To determine the purity, the purified compounds were analyzed by RP-HPLC. The analytical RP-HPLC column was a TOSOH ODS-120T (150 ${\times}$ 4.6 mm i.d, Japan), of which the flow rate was set at 0.7 mL/min using the linear gradient solvent system initiated with 15 % methanol to 85 % methanol for 50 min with monitoring at 254 nm. Under these RP-HPLC conditions, compounds I and II eluted at 3.49 and 4.13 min, respectively. Compound II was found to be most potent and host specific. However, compound I had a unique antibiotic activity against phytopathogenic bacteria like bacterial leaf blight (Xanthomonas oryzae) on rice, where it played a less important role in producing toxicity on T. repens. No toxin activity was detected in the water fraction after partitioning with several organic solvents. However, toxin activity was detected in the ethyl acetate and butanol fractions. In the leaf bioassay using compound II, the disease first appeared within 4-5 h as water soaked rot, which subsequently developed into well-defined blight affecting the whole plant.

Isolation, Purification and Characterization of Antioxidative Bioactive Elastin Peptides from Poultry Skin

  • Nadalian, Mehdi;Kamaruzaman, Nurkhuzaiah;Yusop, Mohd Shakir Mohamad;Babji, Abdul Salam;Yusop, Salma Mohamad
    • Food Science of Animal Resources
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    • v.39 no.6
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    • pp.966-979
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    • 2019
  • Muscle-based by-products are often undervalued although commonly reported having a high amount of natural bioactive peptides. In this study, elastin was isolated from the protein of broiler hen skin while its hydrolysate was prepared using Elastase. Assessment of antioxidative properties of elastin-based hydrolysate (EBH) was based on three different assays; 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical, 2,2'-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical and metal chelating ability. The EBH was purified further using ultrafiltration, gel filtration and Reverse- Phase High-Performance Liquid Chromatography (RP-HPLC). The IC50 of ABTS radical activities for EBH were decreased as EBH further purified using ultrafiltration (EBH III; 0.66 mg/mL)>gel filtration (EB-II; 0.42 mg/mL)>RP-HPLC (EB-II4; 0.12 mg/mL). The sequential identification of the peptide was done by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/ TOF-MS) of the potent fractions obtained from RP-HPLC (EB-II4). The presence of hydrophobic amino acids (Val and Pro) in the peptide sequences could potentially contribute to the high antioxidant activity of EBH. The sequences GAHTGPRKPFKPR, GMPGFDVR and ADASVLPK were identified as antioxidant peptides. In conclusion, the antioxidative potential from poultry skin specifically from elastin is evident and can be explored to be used in many applications such as health and pharmaceutical purposes.

Determination of Bergenin in Different Parts of Bergenia ciliata using a Validated RP-HPLC Method

  • Ali, Ejaz;Hussain, Khalid;Bukhari, Nadeem Irfan;Arshad, Najma;Hussain, Amjad;Abbas, Nasir;Arshad, Sohail;Parveen, Sajida;Shehzadi, Naureen;Qamar, Shaista;Qamar, Abida
    • Natural Product Sciences
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    • v.27 no.1
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    • pp.54-59
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    • 2021
  • Bergenia ciliata (Family: Saxifragaceae) is a folklore remedy for the treatment of various ailments in Asian countries. Bergenin (1) has been isolated as an active constituent in many studies, however, the amount of bergenin has not been determined in all parts of the plant. A simple RP-HPLC method was developed to determine the amount of bergenin in methanol extracts of leaves, rhizomes and roots of the plant. Separation was achieved on an Agilent Eclipse XDB-C18 column maintained at 25 ℃ using isocratic solvent system (water: methanol: acetic acid; 62.5:37:0.5 v/v/v) adjusted at pH 2 0 at a flow rate of 1.0 mL/min. and detected at 275 nm. Correlation coefficient (0.9952) showed linearity of concentration (5-200 ㎍/mL) and response. The values of LOD (0.00947 ㎍/mL) and LOQ (0.02869 ㎍/mL) indicated that method was sensitive. The recovery of bergenin was 99.99-100% indicating accuracy of method. The methanol extract of rhizomes contained higher amount of bergenin (19.4%) than roots (9.2%) and leaves (6.9%). It is concluded that methanol extract of rhizomes is a better source of bergenin than other parts of the plant. The findings are useful for standardization of bergenin containing extracts and herbal preparations.

Sample Preparation for Quantitative Determination of Bisphenols in Canned Beverages Coated with Epoxy by RP-HPLC (RP-HPLC 에 의한 에폭시 통조림관 음료 중 비스페놀류의 분석을 위한 시료의 전처리)

  • Kang, Kyung-Mo;Lim, Heung-Youl;Shin, Hyo-Sun
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1029-1034
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    • 2000
  • A suitable method of sample treatments to minimize the analytical interferences was presented in order to determine bisphenols [bisphenol F (BPF), bisphenol A (BPA), bisphenol F diglycidyl ether (BFDGE), and bisphenol A diglycidyl ether (BADGE)] in various canned beverages coated with epoxy resin by the reversephase high performance liquid chromatography (RP-HPLC) equipped with a fluorescence detector and the gas chromatography with mass selective detection. The recovery test of bisphenols was performed using 1, 5, and 10 ${\mu}g/L$ bisphenols spiked beverages with the combined technique of the solid-phase extraction (SPE) and the liquid-phase extraction (LPE). Both BPA and BADGE showed quite adequate resolutions in HPLC-chromatograms. The recoveries of BPA obtained by LPE with diethyl ether were higher than those obtaind with methylene chloride on coffee, shikhye and fruit juice. For cola and tea, the recoveries of BPA obtaind by SPE were higher than those by LPE with diethyl ether. The recoveries of BADGE were less than those of BPA for all beverage samples treated by either SPE or LPE method. In survey of bisphenols for eighteen commercial canned beverage samples, BPA contents of coffee, cola, tea, shikhye, and fruit juice were in the range of $1.3{\sim}11.6,\;0.5{\sim}0.9,\;1.0{\sim}1.3,\;2.4{\sim}7.9$, and $3.0{\sim}3.4\;{\mu}g/L$, respectively, but there was no detection of BPA in beer sample.

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