• Title/Summary/Keyword: pH-gradient

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Reverse-Phase High Performance Liquid Chromatographic Determination of Amino Acids after Precolumn Derivatization with 1-Dimethyl-aminonaphthalene-5-sulfonyl chloride (Dansyl 유도체화와 역상 고성능액체크로마토그래피에 의한 아미노산의 정량)

  • 나혜경;전덕영;홍윤호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.815-822
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    • 1993
  • This experiment was carried out to determine the separation condition of 1-dimethylaminonaphthalene-5-sulfony(Dansyl) derivatives of amino acids by reverse-phase high performance liquid chromatography with Nova-Pak C18 column. Determined solvent system was solvent A(200mA phosphate buffer pH 6.8 15%, acetonitrile 11%, water 74%) and solvent B(acetonitrile 65%, methanol 28%, water 7%). Linear gradient of solvent B was applied from 12% to 80% for 50min. Complete separation of 20 amino acids including asparagine and glutamine which constitute protein was achieved within 50min. As the detection limit was the range of picomole, the resolution power was excellent. Reproducibility of the retention time was less than mean $\pm$0.05min. According to the above optimum chromatographic conditions, the amino acid composition of some food and human blood was examined. The most affluent amino acid was alanine in human blood, aspartic acid and glutamic acid in soy sauce, alanine and threonine in soy milk and proline in milk and yoghurt.

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Raceway Cultivation of Spirulina platensis Using Underground Water

  • Kim, Choong-Jae;Jung, Yun-Ho;Ko, So-Ra;Kim, Hong-Ik;Park, Yong-Ha;Oh, Hee-Mock
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.853-857
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    • 2007
  • The semi-outdoor cultivation of Spirulina platens is was attempted using an underground-water-based medium. Occurrence of contaminant organisms such as Chlorella sp. and Chlamydomonas sp. was not found from a microscopic observation and bacteria were not detected from denaturing gradient gel electrophoresis(DGGE) analysis of PCR-amplified 16S rDNA during the cultivation, owing to pH control and the high quality of the underground water. The mean productivity was high at $10.5g/m^2/d$ with a range of $4.2-12.3g/m^2/d$ despite the unfavorable weather conditions of the rainy season. The cultivated S. platens is included a normal protein content of 58.9%. Consequently, the underground water improved the biomass productivity and the biomass quality because of an abundant supplementation of natural minerals and through a contaminant-free culture.

Liquid Chromatography-tandem Mass Spectrometry for Quantification of Dioscin in Rat Plasma

  • Kong, Tae Yeon;Ji, Hye Young;Choi, Sang-Zin;Son, Miwon;Lee, Hye Suk
    • Mass Spectrometry Letters
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    • v.4 no.3
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    • pp.55-58
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    • 2013
  • Dioscin is a biologically active steroidal saponin with anticancer and hepatoprotective effects. A rapid, selective, and sensitive liquid chromatographic method with electrospray ionization tandem mass spectrometry was developed for the quantification of dioscin in rat plasma. Dioscin was extracted from rat plasma using ethyl acetate at acidic pH. The analytes were separated on a Halo C18 column using gradient elution of acetonitrile and 0.1% formic acid and detected by tandem mass spectrometry in selected reaction monitoring mode. The standard curve was linear ($r^2$ = 0.998) over the concentration range of 1-100 ng/mL. The lower limit of quantification was 1.0 ng/mL using 50 ${\mu}L$ of plasma sample. The coefficient of variation and relative error for intra- and inter-assay at four QC levels were 1.3 to 8.0% and -5.4 to 10.0%, respectively. This method was applied successfully to the pharmacokinetic study of dioscin after oral administration of dioscin at a dose of 29.2 mg/kg in male Sprague-Dawley rats.

Determination of Aconitine and Related Alkaloids in Processed Buza (부자류 생약의 성분인 아코니틴과 관련 알칼로이드의 정량)

  • 엄동옥;한상욱;신현덕
    • YAKHAK HOEJI
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    • v.44 no.2
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    • pp.135-140
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    • 2000
  • Determination of Aconitum alkaloids in processed Buza (Cho-0, Salted Buza, Moist-heating Buza, Limed Buza), which had been prepared from the raw tubers of Aconitum chiisanenseb(Ranunculaceae), was established using visible spectrophotometry and high-performance liquid chromatography (HPLC) method especially for Aconitine analysis. Aconitum alkloids were reacted with tetra- thiocy-anatocobalt[II] complex ion to form a stable ion pair. The reaction product was insoluble in water but freely soluble in several organic solvents. 1.2-Dichloroethane was the best extracting solvent among the examined solvents. Spectrophotometry of Aconitum alkaloids at nax. 625 was carried out. The HPLC method for aconitine was carried out using Radial PAK-CN column with gradient solvent system by solvent mixture of acetonitrile and phosphate buffer (pH 3.0) at 4$0^{\circ}C$ and 254 nm. Linear relationship was found between absorbance response and concentration of aconitine in range of 0.45 mM~0.9 mM ($r^2$=0.9949) by spectrophotometry and 0.3 mM~1.2mM($r^2$=0.9983) by HPLC method. These methods have been found to be suitable and reproducible for routine analysis of Aconitum alkaloids and its pharmaceutical preparations.

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A comparative study of machine learning methods for automated identification of radioisotopes using NaI gamma-ray spectra

  • Galib, S.M.;Bhowmik, P.K.;Avachat, A.V.;Lee, H.K.
    • Nuclear Engineering and Technology
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    • v.53 no.12
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    • pp.4072-4079
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    • 2021
  • This article presents a study on the state-of-the-art methods for automated radioactive material detection and identification, using gamma-ray spectra and modern machine learning methods. The recent developments inspired this in deep learning algorithms, and the proposed method provided better performance than the current state-of-the-art models. Machine learning models such as: fully connected, recurrent, convolutional, and gradient boosted decision trees, are applied under a wide variety of testing conditions, and their advantage and disadvantage are discussed. Furthermore, a hybrid model is developed by combining the fully-connected and convolutional neural network, which shows the best performance among the different machine learning models. These improvements are represented by the model's test performance metric (i.e., F1 score) of 93.33% with an improvement of 2%-12% than the state-of-the-art model at various conditions. The experimental results show that fusion of classical neural networks and modern deep learning architecture is a suitable choice for interpreting gamma spectra data where real-time and remote detection is necessary.

Simultaneous Liquid Chromatography Tandem Mass Spectrometric Determination of 35 Prohibited Substances in Equine Plasma for Doping Control

  • Kwak, Young Beom;Yu, Jundong;Yoo, Hye Hyun
    • Mass Spectrometry Letters
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    • v.13 no.4
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    • pp.158-165
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    • 2022
  • Many therapeutic class drugs such as beta-blocker, corticosteroids, NSAIDs, etc are prohibited substances in the horse racing industry. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology makes it possible to isolate drugs from interference, enables various drug analyses in complex biological samples due to its sensitive sensitivity, and has been successfully applied to doping control. In this paper, we describe a rapid and sensitive method based on solid-phase extraction (SPE) using solid phase cartridge and LC-MS/MS to screen for different class's 35 drug targets in equine plasma. Plasma samples were pretreated by SPE with the NEXUS cartridge consisted non-polar carbon resin and minimum buffer solvent. Chromatographic separation of the analytes was performed on ACQUITY HSS C18 column (2.1 × 150 mm, 1.8 ㎛). The elution gradient was conducted with 5 mM ammonium formate (pH 3.0) in distilled water and 0.1% formic acid in acetonitrile at a flow rate of 0.25 mL/min. The selected reaction monitoring (SRM) mode was used for drug screening with multiple transitions in the positive ionization mode. The specificity, limit of detection, recovery, and stability was evaluated for validation. The method was found to be sensitive and reproducible for drug screening. The method was applied to plasma sample analysis for the proficiency test from the Association of Racing Chemist.

Exploring the feasibility of Salmonella Typhimurium-specific phage as a novel bio-receptor

  • Choi, In Young;Park, Do Hyeon;Chin, Brayan A.;Lee, Cheonghoon;Lee, Jinyoung;Park, Mi-Kyung
    • Journal of Animal Science and Technology
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    • v.62 no.5
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    • pp.668-681
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    • 2020
  • The purpose of this study was aimed to isolate a Salmonella Typhimurium-specific phage (KFS-ST) from washing water in a poultry processing facility and to investigate the feasibility of the KFS-ST as a novel bio-receptor for the magnetoelastic (ME) biosensor method. KFS-ST against S. Typhimurium was isolated, propagated, and purified using a CsCl-gradient ultracentrifugation. Morphological characteristics of KFS-ST were analyzed using transmission electron microscopy (TEM). Its specificity and efficiency of plating analysis were conducted against 39 foodborne pathogens. The temperature and pH stabilities of KFS-ST were investigated by the exposure of the phage to various temperatures (-70℃-70℃) and pHs (1-12) for 1 h. A one-step growth curve analysis was performed to determine the eclipse time, latent time and burst size of phage. The storage stability of KFS-ST was studied by exposing KFS-ST to various storage temperatures (-70℃, -20℃, 4℃, and 22℃) for 12 weeks. KFS-ST was isolated and purified with a high concentration of (11.47 ± 0.25) Log PFU/mL. It had an icosahedral head (56.91 ± 2.90 nm) and a non-contractile tail (225.49 ± 2.67 nm), which was classified into the family of Siphoviridae in the order of Caudovirales. KFS-ST exhibited an excellent specificity against only S. Typhimurium and S. Enteritidis, which are considered two of the most problematic Salmonella strains in the meat and poultry. However, KFS-ST did not exhibit any specificity against six other Salmonella and 27 non-Salmonella strains. KFS-ST was stable at temperature of 4℃ to 50℃ and at pH of 4 to 12. The eclipse time, latent time, and burst size of KFS-ST were determined to be 10 min, 25 min and 26 PFU/ infected cell, respectively. KFS-ST was relatively stable during the 12-week storage period at all tested temperatures. Therefore, this study demonstrated the feasibility of KFS-ST as a novel bio-receptor for the detection of S. Typhimurium and S. Enteritidis in meat and poultry products using the ME biosensor method.

Gastroesophageal Reflux in Mechanically Ventilated Preterm Infants (기계적 환기 요법이 미숙아 위식도 역류에 미치는 영향)

  • Ku, Hye Jin;Park, Su Eun;Park, Jae Hong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.7 no.1
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    • pp.48-53
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    • 2004
  • Purpose: Besides interferencence of esophageal motor function by the nasogastric tube, a decline of the positive gastro-esophageal pressure gradient caused by intermittent positive pressure ventilation seems to have a major role in the pathogenesis of gastroesophageal reflux (GER) in mechanically ventilated preterm infants. The aim of this study was to determine the incidence of GER and associated risk factors in mechanically ventilated preterm infants. Methods: Twenty four hour esophageal pH monitorings were performed using a antimony electrode on 11 mechanically ventilated preterm infants in Neonatal Intensive Care Unit in Pusan National University Hospital. We evaluated the following reflux parameters; reflux index, reflux episodes/hour, reflux episodes ${\geq}5min/hour$, duration of longest episode, and percent episodes ${\geq}5min$. Patients were considered to have significant GER if more than 2 among 5 parameters were satisfied. Results: The mean gestational age of the patients was 30.9 weeks, mean birth weight was 1,568 g, and mean age at the time of pH monitoring was 2.8 days. Significant GER was detected in 4 patients (36.4%). There was no relationship between the incidence of GER and gestational age, birth weight, postnatal age, or the ventilator settings. Conclusion: The incidence of GER in mechanically ventilated preterm infants was similar, compared with other previous studies. Associated risk factors of GER in these patients were not detected. Therefore, mechanical ventilation in preterm infants does not seem to be the high risk factor of GER.

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Studies on Soybean Protein [Part ll]-Isolation and Subunit Composition of Multiple 7S Globulins- (대두(大豆) 단백질(蛋白質)에 관(關)한 연구(硏究) 제2보[第二報]-7S Globulin중의 복합단백질(複合蛋白質)의 분리(分離) 및 그 구성(構成) Subunit에 대하여)

  • Lee, C.Y.;Kim, I.S.;Kim, S.U.
    • Applied Biological Chemistry
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    • v.20 no.1
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    • pp.26-32
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    • 1977
  • The multiple 7S globulins composed of two fractions (A and B) in the electrophoresis with Davis' method were isolated at different stages of the soybean seed development. Electrophoresis of their subunits liberated in PAWU solvent [phenol-acetic acid-water (2 : 1 : 1) solution plus 5M urea] yielded 4 major bands. Observation of both the electrophoretic bands of the multiple 7S fractions(7S-A and 7S-B) and those of their subunits was suggestive of a similarity of the subunit pattern between two 7S fractions. The two fractions in multiple 7S globulins were isolated with DEAE-Sephadex A-50 column$(2.0{\sim}100cm)$ chromatography. They were separated into 2 fractions in a linear gradient concentration of 0.28 to 0.40M NaCl with phosphate buffer (pH 7.8) containing 10mM ${\beta}-mercaptoethanol$(ME). The isolated protein was dissociated into subunits with two different solvent systems; in PAWU solvent and in Tris-HCl buffer(pH 8.0) containing 1% sodium dodecyl sulfate (SDS) and 40mM ME. The dissociated subunits were subjected to electrophoresis in PAWU-treated 7.5% acrylamide gel and in 1% SDS-treated 5.6% acrylamide gel. In PAWU gel electrophoresis, total 7S globulin was separated into 5 major bands, two of which were occupied in common by two 7S fractions(7S-A and 7S-B). In SDS gel electrophoresis, total 7S globulin was separated into 7 major bands, three of which were overlapped with the subunit of the two 7S fractions. The above results alluded us to the presence of a common and/or similar subunit between the multiple 7S globulins.

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Optimization of Anion-exchange Chromatography for the Separation of Agarase from Culture Broth of Pseudoalteromonas sp. (Pseudoalteromonas sp. 배양액으로부터 아가레이즈 분리를 위한 음이온교환 크로마토그래피 최적화)

  • Kim, Yu-Na;Lee, Jae-Ran;Kim, Mu-Chan;Kim, Sung-Bae;Chang, Yong-Keun;Hong, Soon-Kwang;Kim, Chang-Joon
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.840-845
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    • 2011
  • Degradation products of agarose are biologically active and thus used as an ingredient in pharmaceuticals or functional cosmetics. Furthermore, it has been strongly considered as a substrate for bio-ethanol fermentation. Recently, we isolated new agarase-producing microorganism, Pseudoalteromonas sp. from south sea of Korea. In this study, we aimed to separate and purify the agarase from culture broth of this strain. Separation of agarase was performed by ion- exchange chromatography on DEAE-Sepharose resin. Equilibrium pH and volume ratio of resin to the amount of protein were optimized for the efficient adsorption of protein. 410 ${\mu}g$ of protein was completely adsorbed to 3 mL of resin at pH 7.5. The total amount of eluted protein increased as NaCl concentration increased to 400 mM at isocratic elution. Agarase was separated by linear gradient elution of NaCl (0~1,000 mM). Three major protein peaks were observed and the presence or absence of agarase in these eluted proteins was measured by Lugol's staining technique. Only six eluted protein fractions showed strong agarase activity.