• Title/Summary/Keyword: High Pressure Liquid Chromatography(HPLC)

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Microbiological Degradation of the Phenoxy Herbicide MCPP [2-(2-Methyl-4-Chlorophenoxy) Propionic Acid] (페녹시계 제초제 MCPP [2-(2-Methyl-4-Chlorophenoxy) Propionic Acid]의 미생물학적 분해)

  • Oh, Kye Heon;Olli H. Tuovinen
    • Korean Journal of Microbiology
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    • v.30 no.2
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    • pp.96-100
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    • 1992
  • The microbiological degradation of 2-(2-methyl-4-chloro-phenoxy) propionic acid (MCPP) was evaluated using mixed cultures of soil bacteria. The mixed cultures comprised Pseudomonas species, Flavobacterium species, and Achromobacter species. The bacteria used MCPP as the sole source of carbon and energy but only a partial degradation of the parent compound occurred MCPP degradation proceeded via the formation of 2-methyl-4-chlorophenol (2, 4-MCP) which was detected by high pressure liquid chromatography (PHLC) and confirmed by gas chromatography-mass sepctrometry. This intermediate occurred only transiently and no evidence was seen for the presence of other intermediates detectable by the reverse-phase HPLC or UV absorbance.

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Development of official assay method for loperamide hydrochloride capsules by HPLC

  • Le, Thi-Anh-Tuyet;Nguyen, Bao-Tan;Kim, Min-Ho;Kim, Bit;Kim, Hyun-Soo;Jeong, Seung-Won;Kang, Jong-Seong;Na, Dong-Hee;Chun, In-Koo;Kim, Kyeong Ho
    • Analytical Science and Technology
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    • v.33 no.6
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    • pp.252-261
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    • 2020
  • Currently, the potentiometric titration and the high pressure liquid chromatography (HPLC) method were utilized in Korean Pharmacopoeia XII (KP XII) as well as other pharmacopoeias (USP, EP, BP) for determination of loperamide hydrochloride in raw materials and capsules, respectively. The research objective is to overcome the remaining drawbacks from current methods such as solubility of mobile phase (KP XII), less scientific approach (USP 43) or using paired-ion chromatography reagent which shows some limitations (BP2017 and other formulation monographs). The proposed method was optimized by Design of Experiment (DoE) tool to obtain the satisfied method for determination of loperamide hydrochloride. The optimal condition was performed on the common C18 column (150 mm × 4.6 mm; 5 ㎛) using isocratic elution with the mobile phase containing 40 mM of potassium phosphate monobasic (pH 3.0) and acetonitrile (56:44), at a flow rate of 0.7 mL/min. The optimized method was validated and met the requirements of the International Conference on Harmonization. The developed method was applied to determine loperamide hydrochloride in capsules and can be used to update the current monograph in KP XII.

Ultrasonic-assisted Micellar Extraction and Cloud-point Pre-concentration of Major Saikosaponins in Radix Bupleuri using High Performance Liquid Chromatography with Evaporative Light Scattering Detection

  • Suh, Joon-Hyuk;Yang, Dong-Hyug;Han, Sang-Beom
    • Bulletin of the Korean Chemical Society
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    • v.32 no.8
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    • pp.2637-2642
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    • 2011
  • A new ultrasonic-assisted micellar extraction and cloud-point pre-concentration method was developed for the determination of major saikosaponins, namely saikosaponins -A, -C and -D, in Radix Bupleuri by high performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD). The non-ionic surfactant Genapol X-080 (oligoethylene glycol monoalkyl ether) was chosen as the extraction additive and parameters affecting the extraction efficiency were optimized. The highest yield was obtained with 10% (w/v) Genapol X-080, a liquid/solid ratio of 200:1 (mL/g) and ultrasonic-assisted extraction for 40 min. In addition, the optimum cloud-point pre-concentration was reached with 10% sodium sulfate and equilibration at $60^{\circ}C$ for 30 min. Separation was achieved on an Ascentis Express C18 column (100 ${\times}$ 4.6 mm i.d., 2.7 ${\mu}M$) using a binary mobile phase composed of 0.1% acetic acid and acetonitrile. Saikosaponins were detected by ELSD, which was operated at a $50^{\circ}C$ drift tube temperature and 3.0 bar nebulizer gas ($N_2$) pressure. The water-based solvent modified with Genapol X-080 showed better extraction efficiency compared to that of the conventional solvent methanol. Recovery of saikosaponins ranged from 93.1 to 101.9%. An environmentally-friendly extraction method was successfully applied to extract and enrich major saikosaponins in Radix Bupleuri.

Purification of Antifungal Antibiotic NH-B1 from Actinomycete NH 50 Antagonistic to Plant Pathogenic Fungi (식물병원진균에 길항효과가 있는 방선균 균주 NH50에서 항진균성 항생물질 NH-B1의 순수 분리)

  • 김현겸;김범석;문석식;황병국
    • Korean Journal Plant Pathology
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    • v.14 no.3
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    • pp.191-202
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    • 1998
  • About 300 actinomycetes were isolated from two forest and one sea-shore soil and tested for inhibitory effects on mycelial growth of six plant pathogenic fungi Magnaporthe grisea, Alternaria mali, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f. sp. cucumerinum, and Rhizoctonia solani. Among 300 actinomycetes tested, only 16 actinomycetes showed the antifungal activity against the test fungi. Isolate NH 50 was selected for production and purification of antifungal antibiotic substances. Actinomycete isolate NH 50 displayed the broad antifungal spectra against 11 plant pathogenic fungi. To identify actinomycete isolate NH 50, cultural characteristics on various agar media, diaminopimelic acid type, and morphological characteristics by scanning electron microscopy were examined. As a result, actinomycete isolate NH 50 was classified as a rare actinomycete that had LL-DAP type and did not produce spores. After incubation of isolate NH 50 in yeast extract-malt extract-dextrose broth, antifungal compound NH-B1 that inhibited mycelial growth of some plant pathogenic fungi was purified from the methanol eluates of XAD-16 resins by a series of purification procedures, i.e., silica gel flash chromatography, C18 flash chromatography, Sephadex LH-20 column chromatography, silica gel medium pressure liquid chromatography (MPLC), C18 MPLC, and high pressure liquid chromatography (HPLC). UV spectrum and 1HNMR spectrum of antifungal compound NH-B1 dissolved in methanol were examined. The antibiotic NH-B1 showed the major peaks at 230 and 271.2nm. Based on the data of 1H-NMR spectrum, NH-B1 was confirmed to be an extremely complex polymer of sugars called polysaccharides. The antibiotic NH-B1 showed strong antifungal activity against Alternaria solani and Cercospora kikuchi, but weak activity against M. grisea.

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Detection of Aflatoxins in Soybean Food by HPLC (고속액체 크로마토 그라피에 의한 대두식품중 아플라톡신의 검출)

  • Kim, Young-Kook;Roh, Jung-Koo
    • Korean Journal of Food Science and Technology
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    • v.17 no.4
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    • pp.295-303
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    • 1985
  • Aflatoxin $B_1$, $B_2$, $G_1$, and $G_2$were quantitatively detected by the high pressure liquid chromatography on a Micropak-CN column, with Hexane-THF-IPA-water, using a Lichrosorbpacked flowceil in the fluorometric detector. Under those conditions, the minimum detectable amount of aflatoxin $B_1$ was 0.2 ng. HPLC was used in determining amount of aflatoxins in the commercially manufactured soybean food and home-made Meju. Aflatoxin producing abilities of strains used in the industrially fermented soybean food were also studied with the HPLC technique. Although aflatoxin-like substances were detected in a few samples on TLC, they were not identified with the HPLC retention times of standard aflatoxins. The commercial fungal strains used in Korea had no aflatoxin producing abilities.

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Screening of Inhibitory Effect of Edible Mushrooms on Tyrosinase and Isolation of Active Component (한국산 식용버섯류의 Tyrosinase 활성 저해 검색 및 그 유효성분 분리)

  • 박영현;장성근
    • Journal of Food Hygiene and Safety
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    • v.12 no.3
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    • pp.195-199
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    • 1997
  • For the purpose of isolation and screening of tyrosinase inhibitory activity from edible mushrooms, Pleurotus ostreatus, Auricularia auricula-Judae, Umbilicaria esculenta, Agaricus bisporus, Flammuline velutipes, Lentinus edodes, Ganoderma lucidum, and Coriouls versicolor were examined by tracing inhibitory activities against tyrosinase, utilizing L-3,4-dihydroxyphenylalanine (L-DOPA) as a substrate. Among the eight edible mushrooms tested, Umbilicaria esculenta showed potent enzyme inhibitory activities above 7804% against tyrosinase in ethylacetate (EtOAc) extracts. Ganoderma lucidum and Agaricus bisporus showed inhibitory activities of 67.3% and 51.5% in water extracts. EtOAc extracts of Umbilicaria esculenta was fractionated from silicagel column chromatography and one fraction showed the most inhibitory activity of 60.9%. The three bands (Rf=0.38, 0.27, 0.19) were isolated from preparative TLC of the fraction for purification and identified as mixtures of orsellinate, methyl orsellinate, methyl lecanorate, and methyl gyrophorate by high pressure liquid chromatography (HPLC), ultravisible spectrophotometer (UV), mass spectrophotometer (Mass), nuclear magnetic resonance spectrometer (NMR).

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Consoramides A-C, New Zwitterionic Alkaloids from the Fungus Irpex consors

  • Kim, Ji-Yul;Ki, Dae-Won;Lee, Yoon-Ju;Ha, Lee Su;Woo, E-Eum;Lee, In-Kyoung;Yun, Bong-Sik
    • Mycobiology
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    • v.49 no.4
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    • pp.434-437
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    • 2021
  • In our ongoing search for new secondary metabolites from fungi, a basidiomycete fungus Irpex consors was selected for mycochemical investigation, and three new zwitterionic alkaloids (1-3) and five known compounds (4-8) were isolated from the culture broth (16 l) of I. consors. The culture filtrate was fractionated by a series of column chromatography including Diaion HP-20, silica gel, and Sephadex LH-20, Sep-Pak C18 cartridge, medium pressure liquid chromatography (MPLC), and high pressure liquid chromatography (HPLC) to yield eight compounds (1-8). The structures of the isolated compounds were elucidated by the interpretation of nuclear magnetic resonance (NMR) spectra and high-resolution mass spectrometry (HR-MS). Their antioxidant and antibacterial activities were examined. The zwitterionic structures of three new sesquiterpene alkaloids (1-3) were determined together with five known compounds identified as stereumamide E (4), stereumamide G (5), stereumamide H (6), stereumamide D (7), and sterostrein H (8). This is the first report of the zwitterionic alkaloids in the culture broth of I. consors. Three new zwitterionic alkaloids were named as consoramides A-C (1-3).

Recent Trends of Advanced Biosensors for Mycotoxin Analysis

  • Shim, Won-Bo
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.35-35
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    • 2016
  • A mycotoxin is a toxic secondary metabolite produced by organisms of the fungus kingdom, commonly known as molds and has been widely contaminated in agricultural products such as grains and cereals. Many methods including high performance liquid chromatography (HPLC) and gas chromatography (GC) have already been proposed and reviewed for mycotoxins. These methods are either expensive or time-consuming due to the complication of sample preparation and pre-concentration before determination. In addition, both methods are unsuitable for the routine screening of large sample numbers. A biosensor is a fictive analytical device that combines a biological component with a physicochemical detector for the detection of an analyte. Biosensors represent a rapidly expanding field, at the present time, with an estimated 60% annual growth rate; the major impetus coming from the health-care industry but with some pressure from other areas, such as food safety and environmental monitoring. Antibodies and aptamers are bioreceptors which have been used in the development of biosensors. There are many kinds of antibodies and aptamers specific to mycotoxin, and antibody (or aptamer)-based biosensors have been successfully developed for the detection of mycotoxin. The biosensors permit the rapid, sensitive, simple, and on-site detection of a range of mycotoxins and can be an alternative method to traditional methods such as HPLC and GC. This presentation provides the development trends of biosensors to mycotoxins and their application to food and agricultural products.

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Determination of blood concentration of higenamine by high pressure liquid chromatography

  • Park, Sun-Oak;Hong, Chang-Yee;Paik, Seung-Whan;YunChoi, Hye-Sook
    • Archives of Pharmacal Research
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    • v.10 no.1
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    • pp.60-66
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    • 1987
  • A procedure utilizing high pressure liquid chroatography coupled with UV detection is described for the determination of blood concentration of higenamine. Deproteinized serum was pretreated with$C_{18}$(Sep-pak $C_{18}$ cartridge) and the 70% EtOH eluent was applied onto a reversed-phase column ($\mu$ Bondapak $C_{18}]$) with a 15% acetonitrile in 0.05 N $NaH_2$$PO_4$-trichloroacetic acid mixed buffer (pH 2.8) as a mobile phase. With the UV detection at 232 nm, the retention times of higenamine and 1, 2, 3, 4-tetrahydropapaveroline, an internal standard, were 5.2 min and 3.9 min respectively. The blood concentration of higenamine was meausred at regular intervals after i. v. injection of higenamine to rabbit. A drastic decrease in higenamine concentration to 30% of the maximum value obtained immediately after the injection, was observed during the first 1-2 min period and thereafter the rate of decrease was slowed down. The analytical result seemed to coincide with the pharmacological effect of higenamine exerting the maximum chronotropic and hypotensive effect at the completion of the injections which were progressively recovered.

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Quantification of Karanjin Using High Performance Liquid Chromatography in Raw and Detoxified Karanj (Pongamia glabra vent) Seed Cake

  • Prabhu, T.M.;Devakumar, C.;Sastry, V.R.B.;Agrawa, D.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.3
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    • pp.416-420
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    • 2002
  • Various products of karanj (Pongamia glabra) are utilized for industrial, health and animal agriculture applications in the Indian subcontinent. Despite a rich source of protein (CP, 28-34%), karanj cake was found to be slightly bitter in taste and toxic owing to the presence of flavonoid (Karanjin), restricting its safe inclusion in the livestock diets. Feeding trials with raw cake revealed its poor palatability and adverse performance among different categories of livestock including poultry. The present study was, therefore, aimed to detoxify karanj cake by various physico-chemical methods like solvent extraction, water washing, pressure cooking and alkali and acid treatments. The level of residual karanjin in raw and variously processed cake was quantified using high performance liquid chromatography (HPLC). The raw expeller karanj cake was found to contain about 0.19% of karanjin. Though a non-polar solvent, soxhlet extraction of expeller pressed cake with petroleum ether drastically reduced karanjin content (0.01%). Soaking of cake for 24 h in 1% NaOH (w/w) solution was found to reduce karanjin to a major extent with little further benefit by increasing alkali level. Milder alkalies like lime and fertilizer grade urea reduced the karanjin levels marginally. Similar was the case with mineral acids such as HCl and glacial acetic acid. It was, therefore, concluded that solvent extraction of karanj seeds would be the best method of detoxification as well as for more recovery of oil and karanjin.