• Title/Summary/Keyword: High performance Liquid Chromatography

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Determination of Cholesterol in Milk and Dairy Products by High-Performance Liquid Chromatography

  • Oh, H.I.;Shin, T.S.;Chang, E.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.10
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    • pp.1465-1469
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    • 2001
  • A sensitive high-performance liquid chromatographic method was developed to determine the content of cholesterol in milk and dairy products. To optimize separation of cholesterol, mobile phases including acetonitrile:2-propanol (8:1, v/v), acetonitrile:methanol (3:1, v/v), and acetonitrile:methanoI:2-propanol (7:3: I, v/v/v) were compared. Acetonitrile/methanol/2-propanol was superior to the other mobile phase systems for separating cholesterol. Liquid-liquid extraction (LLE) of cholesterol was simplified using a non-polar solvent, hexane, to remove interfering compounds, and had an excellent recovery $(100{\pm}1.0%)$ of cholesterol. A solid phase extraction (SPE) method using Sep-pak $C_{18}$ was developed and compared with LLE. The SPE method was rapid and highly reproducible. Both extraction methods were useful when used in combination with saponification of esterified cholesterol to facilitate total cholesterol determination. The detection limit of cholesterol was $0.01{\mu}g$. The newly developed HPLC method was rapid, simple, and accurate, and has advantages over the many methods commonly used.

Analysis of DA-6034, a New Flavonoid Derivative in Biological Fluids by HPLC (HPLC를 이용한 생체 시료중의 새로운 플라보노이드 유도체인 DA-6034의 분석)

  • Lee, Jong-Jin;Son, Mi-Won;Yoo, Moo-Hi;Jang, Min-Sun;Kim, Won-Bae;Lee, Kang-Chun
    • YAKHAK HOEJI
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    • v.42 no.2
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    • pp.149-152
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    • 1998
  • A high performance liquid chromatographic method was developed for the determination of DA-6034 in biological fluids using internal standard. Plasma containing DA-6034 and inter nal standard was extracted by liquid-liquid extraction at an acidic pH. After evaporation of the organic layer, the drug and internal standard were reconstituted with mobile phase and injected into the column. They were separated by high performance liquid chromatography on inertsil ODS II column at 334 nm. The detection limit of DA-6034 in plasma was 0.02 ${\mu}$g/ml. In this method, the range of recovery and coefficients of variation were 96-110% and 0.40-3.78%, respectively. There was no interference from endogenous substances. Urine and bile were analysed using the deproteinization method and the detection limit of DA-6034 was 1${\mu}$g/l.

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Quantitation of fructo- and inulo-oligosaccharides by high performance liquid chromatography (High performance liquid chromatography에 의한 fructo 및 inulo올리고당의 정량)

  • Kang, Su-Il;Han, Jong-In;Kim, Kyoung-Youn;Oh, Sun-Jin;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.36 no.4
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    • pp.310-314
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    • 1993
  • High performance liquid chromatographic method using a TSK-gel amide 80 column and isocratic elution with acetonitrile-water (63 :35 ;v/v) mixture was used for the separation and the quantitation of fructo (GF2-GF7)- and inulo-oligosaccharides (F2-F4). Retention time of each standard carbohydrate was highly reproducible. Standardization curves obtained by plotting the peak areas against the amounts of each carbohydrate showed very high coefficient of determination$({\ge}0.9884)$ and similar slopes, and a wide range of y-intercepts. Our results suggest the use of each Pure oligosaccharide for its own standardization curve instead of using a certain carbohydrate as an internal standard.

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Transformation of dissolved organic matter in a constructed wetland: A molecular-level composition analysis using pyrolysis-gas chromatography mass spectrometry

  • Park, Jongkwan;Choi, Mijin;Cho, Jaeweon;Chon, Kyongmi
    • Environmental Engineering Research
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    • v.23 no.4
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    • pp.390-396
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    • 2018
  • This study investigated the transformation of dissolved organic matter (DOM) in a free-water surface flow constructed wetland. Pyrolysis gas chromatography-mass spectrometry (Py-GC/MS) coupled with preparative high-performance liquid chromatography (prep-HPLC) was used to analyze the compositions of biopolymers (polysaccharides, amino sugars, proteins, polyhydroxy aromatics, lipids and lignin) in DOM according to the molecular size at three sampling points of the water flow: inflow, midflow, and outflow. The prep-HPLC results verified the decomposition of DOM through the decrease in the number of peaks from three to one in the chromatograms of the sampling points. The Py-GC/MS results for the degradable peaks indicated that biopolymers relating to polysaccharides and proteins gradually biodegraded with the water flow. On the other hand, the recalcitrant organic fraction (the remaining peak) in the outflow showed a relatively high concentration of aromatic compounds. Therefore, the ecological processes in the constructed wetland caused DOM to become more aromatic and homogeneous. This indicated that the constructed wetland can be an effective buffer area for releasing biochemically stable DOM, which has less influence on biological water quality indicators, e.g., biochemical oxygen demand, into an aquatic ecosystem.

Determination of Patulin in Commercial Apple Juice in Korea by High Performance Liquid Chromatography (High Performance Liquid Chromatography를 이용한 국내 시판 사과 주스의 Patulin 분석)

  • Cho, Wan-Il;Choi, Young-Boong;Moon, Tae-Wha
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.412-416
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    • 1997
  • Seven brands of commercial apple juice in Korea obtained from local stores were analyzed for patulin. Sep-Pak silica cartridge purification of the extract by ethyl acetate offered a good separation of patulin from other components. Patulin was determined by reverse phase liquid chromatography using a J'sphere $4\;{\mu}m$ ODS-H80 column with an ultraviolet detector set at 274 nm. Patulin concentrations in four samples ranged from 4 to $20\;{\mu}g/L$ and the other three samples from 30 to $45\;{\mu}g/L$. The limit of detection was $3\;{\mu}g/L$, and the recovery was $80{\sim}90%$ at the contamination level of $3.78{\sim}125.9\;{\mu}g/L$.

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Separation and Purification of Lipase Inhibitory Peptide from Fermented Milk by Lactobacillus plantarum Q180

  • Kim, Seulki;Lim, Sang-Dong
    • Food Science of Animal Resources
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    • v.40 no.1
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    • pp.87-95
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    • 2020
  • In this study, we separated and purified lipase inhibitory peptide from fermented milk by Lactobacillus plantarum Q180 with the aim of developing a new functional anti-lipase activity yogurt product. L. plantarum 180 was inoculated into 10% reconstituted skimmed milk and incubated at 37℃ until the pH of the culture reached pH 4.4. The lipase activity was measured using porcine pancreatic lipase. The lipase inhibitory peptides were gradually isolated by ultrafiltration, reversed phase column chromatography (RPC), reversed phase high-performance liquid chromatography (RP-HPLC), and gel permeation high-performance liquid chromatography (GP-HPLC) from the fermented milk by L. plantarum Q180. An ODS-AQ column was used for the RPC, a Vydac C18 column for the RP-HPLC, and a Superdex Peptide HR column for the GP-HPLC. The peptide was composed of Asp, Thr, Ile, Ser, Ala, and Gln, and the anti-lipase activity (IC50) was 2,817 ㎍/mL.

Identification and Quantitative Analysis of Flavonol Glycosides from Ginkgo biloba Leaves by High Performance Liquid Chromatography (고속액체크로마토그라피에 의한 은행잎중 Flavonoid Glycoside의 확인 및 정량)

  • Kang, Sam-Sik;Kim, Ju-Sun;Kwak, Wie-Jong;Kim, Ki-Hyup
    • Korean Journal of Pharmacognosy
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    • v.21 no.2
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    • pp.148-152
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    • 1990
  • Seven flavonol glycosides from the EtOAc fraction of Ginkgo biloba leaves were identified by high performance liquid chromatography. Separation by reversed phase chromatography on $Lichrosorb^{\circledR}$ RP-18 column was achieved by isocratic elution. The content of the major acylated flavonol glycoside, kaempferol 3-0-[$6'-O-{p}-coumaroyl-{\beta}-_D-glucosyl(1{\rightarrow}2)-{\alpha}-_L-rhamnoside$] was about 8.0% and 0.55% for EtOAc fraction and MeOH extract, respectively.

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Partial Purification of the Outer Membrane-Associated 2-Furaldehyde Dehydrogenase from Klebsiella pneumoniae (Klebsiella pneumoniae 균주의 세포외막으로부터 2-Furaldehyde Dehydrogenase의 부분정제에 관하여)

  • 이준우;이병웅;강사욱;하영칠;유병설;한홍의
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.370-376
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    • 1986
  • From the outer membrane portion of Gram-negative Klebsiella pneumoniae, the activity of 2-furaldehyde dehydrogenase depending upon beta-nicotinamide adenine dinucleotide was detected. Cytoplasmic membrane was preferentially extracted from crude membrane with $Mg^{2+}$ and Triton X-100, and then outer membrane was collected by ultracentrifugation. The crude enzyme was obtained by solubilization of outer membrane with lysozyme, ethylene diamine tetraacetate and Triton X-100. Thereafter 2-furaldehyde dehydrogenase was partially purified through column chromatography on QAE-Sephadex Q-50 and Sephadex G-150 and the enzyme activity was analyzed by means of high performance liquid chromatography. The optimal pH for the activity of the enzyme was about 9.5 and the optimal temperature was about $85^{\circ}C$. The partially purified enzyme retained tis activity at $85^{\circ}C$ for 5 hours. The optimal concentration of Triton X-100 for the activity of the enzyme was about 1.5% in the reaction mixture.

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Identification of Actinomycins by High Performance Liquid Chromatography and Fast Atom Bombardment Mass Spectrometry

  • Cho, Seong-Eun;Goo, Yang-Mo;Kim, Kyoung-Ja
    • Archives of Pharmacal Research
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    • v.17 no.6
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    • pp.424-427
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    • 1994
  • An acinomycin complex isolated from culture broth of a soil microorganism, SNUS 9305-011 has been examined by High performance liquid chromatography (HPLC). From the analysis of the fractions obtained by column chromatography of the ethyl acetate extract, three actinomycin components are confimed . The HPLC analysis is carried out with a CN-bonded nucleosil column. Comparison of the retention times of the components with those of actinomycin D, C complex, $X_{o{\beta}$, and V and suggests that they are different actinomycins. FBA mass spectra fo the coponents also shows different molecular ions from those of standards and other reported actionbmycins. The present work has demonstrated that actinomycin components can be separated by a CN-bonded HPLC column, and that ocmparison of their HPLC chormatograms with authentic smaples and information on their molecular ions can be successfully employed for indentification of actionmycins.

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Determination of Fluorescent Whitening Agents in Paper Materials by Ion-Pair Reversed-Phase High-Performance Liquid Chromatography

  • Kim, Jeong Soo;Kim, Do Hwan;Kim, Keon
    • Bulletin of the Korean Chemical Society
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    • v.33 no.12
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    • pp.3971-3976
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    • 2012
  • A simple method was developed for the analysis of seven stilbene-type fluorescent whitening agents (FWAs) in paper materials by ion-pair reversed-phase high-performance liquid chromatography with fluorescence detection. These stilbene-type FWAs included two disulfonate, two tetrasulfonate, and three hexasulfonate compounds. After optimization of chromatographic conditions, the FWAs were satisfactorily separated using a reversed-phase column (RP-18) with the following isocratic mobile phase: methanol-water (60:40) containing 17.5 mM TBABr and 10 mM citrate buffer (pH = 7.0). The calibration plot was linear in the range from 5 to 500 ng/mL for two disulfo-FWAs and from 1 to 500 ng/mL for the other five FWAs. Precision levels of the calibration curve as indicated by RSD of response factors were 1.2 and 8.1%. Limits of quantitation (LOQ) ranged from 1.2 to 11 ng/mL.