• Title/Summary/Keyword: Liquid Liquid Extraction

Search Result 1,075, Processing Time 0.028 seconds

Development of ELISA for Brucella abortus RB51 II. Purification of 8kDa antigen and development of ELISA using its antigen of Brucella abortus RB51 (부루세라 RB51의 ELISA 진단법 개발 II. Brucella abortus RB51균의 8kDa 항원 정제 및 ELISA 진단법 개발)

  • Her, Moon;Cho, Dong-hee;Jung, Byeong-yeal;Cho, Seong-kun;Jung, Suk-chan;Kim, Ok-kyung
    • Korean Journal of Veterinary Research
    • /
    • v.41 no.1
    • /
    • pp.51-57
    • /
    • 2001
  • A procedure for extraction and purification of 8 kDa antigen of Brucella abortus RB51 was developed. Bacteria heat inactivated at $60^{\circ}C$, 30 min was extracted by 1% sarcosine and followed by fluid pressure liquid gel filtration chromatography of 2 series, Superose 12 HR 10/30 and Sephacryl S-100. There was produced $71.46{\mu}g/g$(wet) of 8 kDa antigen, and it resisted 1% trypsin, solved 1% triton X-100 higher than distilled water and inactivated 0.1% proteinase K. These results show that 8 kDa antigen may be a lipoprotein existed cell surface of B. abortus RB51. Also, we developed ELISA using purified 8 kDa surface antigen of Brucella abortus RB51 strain, its specificity and sensitivity was 95.0%, 98.6%, respectively. As compared with dot-blot assay using whole cell and ELISA using 8 kDa antigen, its correlation was 93.5%.

  • PDF

Simultaneous Determination of the Flavonoids and Limonoids in Citrus junos Seed Shells Using a UPLC-DAD-ESI/MS

  • Jo, Ara;Shin, Ji hun;Song, Hwa young;Lee, Ye Eun;Jeong, Da Eun;Oh, Sung Hwa;Mun, Myung Jae;Lee, Mina
    • Natural Product Sciences
    • /
    • v.26 no.1
    • /
    • pp.64-70
    • /
    • 2020
  • Citrus junos seeds (CS) have been traditionally used for the treatment of cancer and neuralgia. They are also used to manufacture edible oil and cosmetic perfume. A large amount of CS shells without oil (CSS) are discarded after the oil in CS is used as foods or herbal remedy. To efficiently utilize CSS as a by-products, it needs to be studied through chemical analysis. Therefore, we developed an ultra-performance liquid chromatography (UPLC)-diode array detection (DAD) method for simultaneous determination and quantitative analysis of five components (two flavonoids and threes limonoids) in CSS. A Waters Acquity UPLC HSS T3 column C18 (2.1 × 100 mm, 1.8 ㎛) was used for this separation. It was maintained at 40 ℃. The mobile phase used for the analysis was distilled water and acetonitrile with gradient elution. To identify the quantity of the five components, a mass spectrometer (MS) with an electrospray ionization (ESI) source was used. The regression equation showed great linearity, with correlation coefficient ≥ 0.9912. Limits of detection (LOD) and limits of quantification (LOQ) of the five compounds were 0.09 - 0.13 and 0.26 - 0.38 ㎍/mL, respectively. Recoveries of extraction ranged from 97.45% to 101.91%. Relative standard deviation (RSD) values of intra- and inter-day precision were 0.06 - 1.15% and 0.19 - 0.25%, respectively. This UPLC-DAD method can be validated to simultaneously analyze quantities of marker flavonoids and limonoids in CSS.

Recovery of C-14 in the Cement Waste Form (농축폐액 시멘트 고화체로부터 C-14 회수 특성)

  • Ahn Hong-Joo;;Lee Jeong-Jin;Pyo Hyung-Yeal;Han Sun-Ho;Jee Kwang-Young
    • Proceedings of the Korean Radioactive Waste Society Conference
    • /
    • 2005.06a
    • /
    • pp.284-289
    • /
    • 2005
  • According to the nuclear safety regulation policy including the administration of radionuclides in low level radwastes, the evaporator bottoms were mixed with cement to form a stable solidification for identifying the recovery possibility of the C-14. The chemical oxidation method was applied for the extraction of C-14 from the cement waste form. The emitting beta ray of the C-14 extracted from the radwastes was measured with the liquid scintillation counter and calculated by using the quenching correction curves. Only the beta emitting radioactive nuclides of the C-14 in the radwastes was showed the radioactivities with the range of $2.7E+00\;{\sim}\;3.07E+02$ Bq/g.

  • PDF

Analysis of Estrogen in Pomegranate Extract by Solid Phase Extraction and Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS를 이용한 석류추출물 중의 에스트로겐 분석)

  • Kum, Eun-Joo;Kwon, Do-Hyeong;Shin, Hye-Seoung
    • Journal of Food Hygiene and Safety
    • /
    • v.25 no.1
    • /
    • pp.79-82
    • /
    • 2010
  • The pomegranate (Punica granatum), especially its fruit, possesses a vast ethnomedical history and represents a phytochemical reservoir of heuristic medical value. The tree and fruit can be divided into several anatomical compartments, and the fruit juice, peel and oil are known to be weakly estrogenic and heuristically of interest for treatment of menopausal symptoms and sequellae. In this study, analysis of estrogen in pomegranate extract was carried out with LC/MS/MS. Three batches of pomegranate extract samples were used to analysis the target compounds (estrogen). The contents of estrogen derivatives in the samples were 38.6 ppb of estriol, 83.5 ppb of estrone, and 10.9 ppb of estradiol. This result suggests that the pomegranate extract can used for treatment of menopause symptoms in the woman.

Establishment of an Analytical Method of Fluoroquinolones in Milk by HPLC (HPLC를 이용한 우유에서의 fluoroquinolones 시험법 확립)

  • Kim, Jong-Hwa;Hong, Se-Lyung;Kang, Tae-Beom;Lee, Hyun-Kyung;Lee, Soon-Ho
    • Korean Journal of Food Science and Technology
    • /
    • v.42 no.5
    • /
    • pp.521-526
    • /
    • 2010
  • A high-performance liquid chromatography (HPLC) method was established for the determination of fluoroquinolones in milk. Protein was removed by using trichloroacetic acid in order to increase a mean recovery of milk. The extracts were using $Strata^{TM}$-X solid-phase extraction cartridge. The analytes were detected by HPLC on a $C_{18}$ column. HPLC method with fluorescence detection system (Ex: 278 nm, Em: 456 nm) provided a high degree of sensitivity in detecting fluoroquinolones. The limits of quantitation (LOQ) and mean recoveries of fluoroquinolones were 40 ${\mu}g$/kg and 73.6-95.2% (ofloxacin), 10 ${\mu}g$/kg and 77.3-91.9% (norfloxacin), 20 ${\mu}g$/kg and 91.6-94.3% (ciprofloxacin), 10 ${\mu}g$/kg and 81.0-87.8% (enrofloxacin), 10 ${\mu}g$/kg and 71.3-81.0% (sarafloxacin), 10 ${\mu}g$/kg and 89.4-90.8% (orbifloxacin), 2 ${\mu}g$/kg and 69.4-85.5% (danofloxacin).

Isolation of Eckol from Ecklonia cava via Centrifugal Partition Chromatography (CPC) and Characterization of it's Anti-inflammatory Activity (고속원심분배 크로마토그래피를 이용한 감태(Ecklonia cava)로부터 Eckol의 분리 및 항염증 활성)

  • Kim, Yoon Taek;Lee, Ji-Hyeok;Ko, Ju-Young;Oh, Jae-Young;Lee, Won-U;Sok, Chang Hyun;Hong, Jin Tae;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.48 no.3
    • /
    • pp.301-307
    • /
    • 2015
  • Phlorotannins and marine algal polyphenols, including dieckol, 6,6-bieckol, phloroglucinol, phlorofucofuroeckol-A, and eckol, were isolated from brown seaweeds. These compounds have beneficial bioactivities, and Ecklonia cava has become widely used for the extraction and isolation of phlorotannins. Eckol, in particular, has been to shown to have antioxidant, anti-inflammatory, anticoagulatory, and photoprotective properties. However, due to its low abundance in weaweed, the isolation and purification of eckol are difficult. Its limited availability renders the isolation and purification of eckol labor-intensive processes. Centrifugal partition chromatography (CPC) is an efficient technique for the isolation and purification of eckol. In this study, eckol was isolated from the ethyl acetate fraction of the 70% ethanol extract of E. cava using CPC with a two-phase solvent system of a n-hexane:EtOAc:methanol:water (2:8:3:7, v/v) solution. The purity and anti-inflammatory activity of the isolated eckol were verified by high-performance liquid chromatography and by assaying lipopolysaccharide-induced inflammatory responses in an immortalized murine BV2 microglial cell line, respectively. In conclusion, CPC is a useful technique for simple and efficient isolation of eckol from E. cava.

Anitoxidative components of pueraria root (칡뿌리의 항산화 성분)

  • Oh, Man Jin;Lee, Ka Soon;Son, Hwa Young;Kim, Seung Yeol
    • Korean Journal of Agricultural Science
    • /
    • v.17 no.1
    • /
    • pp.52-64
    • /
    • 1990
  • As a search for natural antioxidants, antioxdative fractions in Pueraria roots were extracted, identified using column chromatography, thin layer chromatography or high performance liquid chromatography. The components which have most effective antioxidative activities were further identified by IR and GC-MS. Separated antioxidative components were then added to four different oils to examine their antioxidative activities. Yield of extract obtained from pueraria root powder by solvent extraction using four step solvent systems was 2.54%. Antioxidative activity of the extracts was as effective as that of 100 ppm ${\delta}$-tocopherol addition, when 0.1% of the extracts was added to linoleic acid. The strongest antioxidative component of methanol extract of pueraria root was identified as puerarin. Aunioxidative activity of puerarin on lard was more effective than ${\alpha}$-tocopherol, but less effective than ${\delta}$-tocopherol. When the puerarin was added to edible oil and heat treated at $145^{\circ}C$, the acid value was lowest in lard and was highest in soybean oil. Antioxidative activity in terms of carbonyl value, thiobarbituric acid value and anisidine value was most high in palm oil and least in soybean and cottonseed oil.

  • PDF

Ginsenosides analysis of New Zealand-grown forest Panax ginseng by LC-QTOF-MS/MS

  • Chen, Wei;Balan, Prabhu;Popovich, David G.
    • Journal of Ginseng Research
    • /
    • v.44 no.4
    • /
    • pp.552-562
    • /
    • 2020
  • Background: Ginsenosides are the unique and bioactive components in ginseng. Ginsenosides are affected by the growing environment and conditions. In New Zealand (NZ), Panax ginseng Meyer (P. ginseng) is grown as a secondary crop under a pine tree canopy with an open-field forest environment. There is no thorough analysis reported about NZ-grown ginseng. Methods: Ginsenosides from NZ-grown P. ginseng in different parts (main root, fine root, rhizome, stem, and leaf) with different ages (6, 12, 13, and 14 years) were extracted by ultrasonic extraction and characterized by Liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Twenty-one ginsenosides in these samples were accurately quantified and relatively quantified with 13 ginsenoside standards. Results: All compounds were separated in 40 min, and a total of 102 ginsenosides were identified by matching MS spectra data with 23 standard references or published known ginsenosides from P. ginseng. The quantitative results showed that the total content of ginsenosides in various parts of P. ginseng varied, which was not obviously dependent on age. In the underground parts, the 13-year-old ginseng root contained more abundant ginsenosides among tested ginseng samples, whereas in the aboveground parts, the greatest amount of ginsenosides was from the 14-year-old sample. In addition, the amount of ginsenosides is higher in the leaf and fine root and much lower in the stem than in the other parts of P. ginseng. Conclusion: This study provides the first-ever comprehensive report on NZ-grown wild simulated P. ginseng.

Studies on the Development of Liquid Chromatographic Methods for Pesticide Residues (II) : The Development of the Analytical Method for Thiocarbamates Herbicides (잔류농약의 액체 크로마토그래피 분석법 개발에 관한 연구 (II) Thiocarbamates 제초제의 잔류농약 분석법 개발)

  • Lee Dai Woon;Choi Yong Wook
    • Journal of the Korean Chemical Society
    • /
    • v.36 no.1
    • /
    • pp.66-74
    • /
    • 1992
  • A method for the multiresidual simultaneous analysis of 11 thiocarbamates was studied using HPLC. Thiocarbamate in Chinese cabbage was analyzed in the order of extraction, partition, and cleanup in their optimum condition. Acetone was chosen as an extracting solvent. As a partitioning solvent, the mixture of 50% methylene chloride and petroleum ether containing extremely small water content showed good recoveries of thiocarbamate from the water layer. Partition efficiency was affected by pH of the water layer; it remained almost constant under the acidic and neutral condition while decreasing under the basic condition. The comparison done in cleanup step showed that the column chromatographic method is superior to the treatment of coagulating reagent. As an absorbent, the mixture of charcoal, magnesia, and celite with the ratio of 1 : 2 : 4 gave better recoveries and also effectively removed chlorophyll. Over the total procedure, the average recoveries for thiocarbamates in Chinese cabbage were 91% at about 2 ppm fortification level within the relative standard deviation of 8%, and the minimum detection limit (MDL) was 2.2${\sim}$9.3 ng.

  • PDF

Development of a Quantitative Analytical Method for Determining the Concentration of Human Urinary Paraben by LC-MS/MS

  • Lee, Seung-Youl;Son, Eunjung;Kang, Jin-Young;Lee, Hee-Seok;Shin, Min-Ki;Nam, Hye-Seon;Kim, Sang-Yub;Jang, Young-Mi;Rhee, Gyu-Seek
    • Bulletin of the Korean Chemical Society
    • /
    • v.34 no.4
    • /
    • pp.1131-1136
    • /
    • 2013
  • Parabens, the esters of p-hydroxybenzoic acid, have been widely used as antimicrobial preservatives in cosmetic products, drugs, and processed foods and beverages. However, some parabens have been shown to have weak estrogenic effects through in vivo and in vitro studies. Because such widespread use has raised concerns about the potential human health risks associated with exposure to parabens, we developed a simultaneous analytical method to quantify 4 parabens (methyl, ethyl, propyl, and butyl) in human urine, by using solid-phase extraction and high-performance liquid chromatography coupled with triple quadrupole mass spectrometry. This method showed good specificity, linearity ($R^2$ > 0.999), accuracy (92.2-112.4%), precision (0.9-9.6%, CV), and recovery (95.7-102.0%). The LOQs for the 4 parabens were 1.0, 0.5, 0.2, and 0.5 ng/mL, respectively. This method could be used for quick and accurate analysis of a large number of human samples in epidemiological studies to assess the prevalence of human exposure to parabens.