• Title/Summary/Keyword: Fluorescence Spectrometry

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Purification, and Biochemical and Biophysical Characterization of Cellobiohydrolase I from Trichoderma harzianum IOC 3844

  • Colussi, Francieli;Serpa, Viviane;Da Silva Delabona, Priscila;Manzine, Livia Regina;Voltatodio, Maria Luiza;Alves, Renata;Mello, Bruno Luan;Nei, Pereira Jr.;Farinas, Cristiane Sanches;Golubev, Alexander M.;Santos, Maria Auxiliadora Morim;Polikarpov, Igor
    • Journal of Microbiology and Biotechnology
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    • v.21 no.8
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    • pp.808-817
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    • 2011
  • Because of its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum has a considerable potential in biomass hydrolysis applications. Trichoderma harzianum cellobiohydrolase I (ThCBHI), an exoglucanase, is an important enzyme in the process of cellulose degradation. Here, we report an easy single-step ion-exchange chromatographic method for purification of ThCBHI and its initial biophysical and biochemical characterization. The ThCBHI produced by induction with microcrystalline cellulose under submerged fermentation was purified on DEAE-Sephadex A-50 media and its identity was confirmed by mass spectrometry. The ThCBHI biochemical characterization showed that the protein has a molecular mass of 66 kDa and pI of 5.23. As confirmed by smallangle X-ray scattering (SAXS), both full-length ThCBHI and its catalytic core domain (CCD) obtained by digestion with papain are monomeric in solution. Secondary structure analysis of ThCBHI by circular dichroism revealed ${\alpha}$- helices and ${\beta}$-strands contents in the 28% and 38% range, respectively. The intrinsic fluorescence emission maximum of 337 nm was accounted for as different degrees of exposure of ThCBHI tryptophan residues to water. Moreover, ThCBHI displayed maximum activity at pH 5.0 and temperature of $50^{\circ}C$ with specific activities against Avicel and p-nitrophenyl-${\beta}$-D-cellobioside of 1.25 U/mg and 1.53 U/mg, respectively.

Determination of polycyclic aromatic hydrocarbons (PAHs) in used lubricating car oils (차량용 폐윤활유에 함유된 다환 방향족 탄화수소 (PAHs)의 분석)

  • Yoo, Kwang-Sik;Jyoung, Ji-Young;Jeong, Seon-Yi;Woo, Sang-Beom
    • Analytical Science and Technology
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    • v.16 no.5
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    • pp.339-348
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    • 2003
  • Determination of some PAHs in used engine oils have been carried out by extraction of the components into acetonitrile followed by GC/FID and synchronous spectrofluorimetric technique. 7 PAHs, such as acenaphthene (Ace), anthracene (Anth), benzo(a)pyrene (BaP), chrysene (Chry), phenanthrene (Phen), fluoranthene (Ft), and perlyrene (Per) in used engine oil sample were able to determine separately by synchronous spectrofluorimetry. Calibration curves for those components were linear for the concentration range of 0.4~166 ppb PAHs with the corelation factor of 0.9985~0.9999. The peak areas produced by GC/FID split ratio program were used for the calibration curves of the other 8 PAHs. Detection sensitivity of the synchronous spectrofluorimetry seems to be 100 times more sensitive than GC/FID method. The total amount of PAHs in the used engine oil were 5.5 ng/g for LNG (bus), 10.5 ng/g for LPG(taxi), 92.2 ng/g for gasoline-passenger car, and 130 ng/g for diesel trailer, respectively.

Spectmscopic and Magnetic Properties of Yanggiseok, Yeonok and Eumgiseok used as Mineral Medicine (광물성 한약으로 이용되는 양기석, 연옥,음기석의 분광학적 및 자기적 특성)

  • 김선옥;박맹언;정율필
    • Economic and Environmental Geology
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    • v.35 no.4
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    • pp.317-323
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    • 2002
  • Mineral medicines are single or mixtures of minerals and rocks which have been used to treat disease. Recently, their application has been increased by emphasizing the physical properies of the medicines, in addition to their chemical properies. In this study, mineralogical, chemical, spectroscopic and magnetic properties were measured using X-ray diffraction analysis, X-ray fluorescence spectrometry, Ff-Infrared spectroscopy, nuclear magnetic resonance spectroscopy, mass magnetic susceptibility. Experiments were done using these properties to evaluate application of traditional mineral medicines such as Yanggiseok, Yeonok and Eumgiseok. Mineralogical study proves that Yanggiseok, generally known as tremolite, consists of actinolite. Yeonok for medical usage mostly consists of fine grained tremolite. Eumgiseok is mainly composed of vermiculite and minor kaolinite and halloysite. Yeonok and Yanggiseok, belong to the amphibole group among inosilicates and both have similar emission power properties. The intensity of emission power, calculated from FT-IR measurements, follows in the order of Yeonok, Yanggiseok and Eumgiseok at 40$^{\circ}$C and Yanggiseok, Yeonok and Eumgiseok in such order at 150$^{\circ}$C. As a result of NMR analysis after 20 days in distilled water, the three mineral medicines decreased in the following order; Eumgiseok, Yanggiseok and Yeonok. However, the same minerals decreased in the order of Eumgiseok, Yeonok and Yanggiseok after 80 days. In response temperature, magnetic susceptibility of Yanggiseok and Eumgiseok systematically increased by heating to 25$^{\circ}$C, 100$^{\circ}$C and 700$^{\circ}$C. Magnetic susceptibility of Yeonok shows a decreasing pattern due to heating.

A Study on Chemical Composition and Firing Temperature of White Wares from the Guyre-2ri Kiln Sites, Wonju (원주 귀래2리 가마터 출토 백자의 화학조성과 소성온도 연구)

  • Lee, Byeong Hoon;So, Myoung-Gi
    • Journal of Conservation Science
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    • v.30 no.3
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    • pp.249-261
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    • 2014
  • This study aims to examine production technique of white wares from the Guyre-2ri 1st and 2nd kiln sites, Wonju, Gangwon Province and characteristics of the used materials, and to find a correlation among materials of the excavated white wares. X-ray fluorescence sequential spectroscopy(XRF), X-ray diffraction(XRD), Dilatometer and Inductively coupled plasma mass spectrometry(ICP-MS), Inductively coupled plasma automic emission spectrometer(ICP-AES) were applied to determine the chemical composition, crystalline phase of samples, firing temperatures, trace elements and rare earth elements. White wares from the Guyre-2ri kiln sites contained high contents of coloring oxides and fluxes. Though firing temperature of each sample was different, they were mostly fired at a temperature below $1200^{\circ}C$ and some of them experienced a low temperature of $1000^{\circ}C{\pm}20^{\circ}C$ and a high temperature above $1200^{\circ}C$. When analyzing body crystalline phases of the white wares using the XRD method, quartz and mullite were extracted from all the samples, and the proportions were similar to each other. When analyzing the excavated white wares using the Seger formula, also, all the samples showed similar clay sources and production techniques. Moreover, the white wares were made of host rocks of the same geological origin, according to the result of rare earth elements analysis.

Analysis of Methamphetamine and Amphetamine in Oral Fluid of Eleven Drug Abusers (마약남용자 11명의 타액 중 메스암페타민의 분석)

  • Kim, Eun-Mi;Lee, Ju-Seon;Choi, Hye-Young;Choi, Hwa-Kyung;Chung, Hee-Sun
    • YAKHAK HOEJI
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    • v.52 no.6
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    • pp.419-425
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    • 2008
  • A qualitative and quantitative analytical method was developed for detection of methamphetamine (MA) and its main metabolite amphetamine (AM) in oral fluid. Oral fluids of eleven drug abusers were provided by Police, specimens were collected by stimulation with a cotton swab treated with 20 mg of citric acid ($Salivette^{(R)}$; Sarstedt, USA). As the preliminary test, oral fluid samples were screened for amphetamines by Fluorescence Polarization Immunoassay (TDxFLx, Abbott Co.). Extraction for MA was performed using solid-phase extraction (SPE) by $RapidTrace^{TM}$ (Zymark, USA) with mixed mode cation exchange cartridge, CLEAN $SCREEN^{(R)}$ (130 mg/3 ml, UCT) after dilution with phosphate buffer. Samples were evaporated and derivatized by pentafluoropropionic acid anhydride (PFPA). Quantitation of MA and AM was performed by gas chromatography-mass spectrometry (GC-MS) using selective ion monitoring (SIM), the quantitation ions were m/z 204 (MA), 208 (MA-$D_5$), 190 (AM) and 194 (AM-$D_5$). The selectivity, linearity of calibration, limit of detection (LOD) and quantification (LOQ) within- and between day precision, accuracy and recoveries were examined as parts of the method validation. All oral fluid samples gave positive results to immunoassay for MA (cut-off level, 50 ng/ml as d-amphetamine). Concentrations of MA and AM by GC-MS in eleven samples were ranged 104.2${\sim}$4603.3 ng/ml and 32.4${\sim}$268.6 ng/ml, respectively. Extracted calibration curves of MA and AM were linear over the two concentration range of 1${\sim}$100 and 50${\sim}$1000 ng/ml with correlation coefficient of above 0.999. LOQ of MA and AM was 1 and 3 ng/ml, respectively. The intraand inter-day run precisions (CV) for MA and AM were less than 10%, and the accuracies (bias) for MA and AM were also less than 10% at the two different concentrations 5 and 100 ng/ml at low calibration range, 50 and 1000 ng/ml at high calibration range. The absolute recoveries of MA and AM at low and high calibration ranges were more than 82% and 75%, respectively. In this study the qualitative and quantitative analytical method of MA in oral fluid was established. Oral fluid testing may detect drug use in past hours because of its shorter detection window than urine, and be useful in post-accident situations. So oral fluids will be most useful for testing drug abuse in the driving under the influence of drug (DUID) as the alternative specimens of urine.

Studies on the Analysis of Benzo(a)pyrene and Its Metabolites on Biological Samples by Using High Performance Liquid Chromatography/Fluorescence Detection and Gas Chromatography/Mass Spectrometry

  • Lee, Won;Shin, Hye-Seung;Hong, Jee-Eun;Pyo, Hee-Soo;Kim, Yun-Je
    • Bulletin of the Korean Chemical Society
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    • v.24 no.5
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    • pp.559-565
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    • 2003
  • An analytical method the determination of benzo(a)pyrene (BaP) and its hydroxylated metabolites, 1-hydroxybenzo(a)pyrene (1-OHBaP), 3-hydroxybenzo(a)pyrene (3-OHBaP), benzo(a)pyrene-4,5-dihydrodiol (4,5-diolBaP) and benzo(a)pyrene-7,8-dihydrodiol (7,8-diolBaP), in rat urine and plasma has been developed by HPLC/FLD and GC/MS. The derivatization with alkyl iodide was employed to improve the resolution and the detection of two mono hydroxylated metabolites, 1-OHBaP and 3-OHBaP, in LC and GC. BaP and its four metabolites in spiked urine were successfully separated by gradient elution on reverse phase ODS $C_{18}$ column (4.6 mm I.D., 100 mm length, particle size 5 ㎛) using a binary mixture of MeOH/H₂O (85/15, v/v) as mobile phase after ethylation at 90 ℃ for 10 min. The extraction recoveries of BaP and its metabolites in spiked samples with liquid-liquid extraction, which was better than solid phase extraction, were in the range of 90.3- 101.6% in n-hexane for urine and 95.7-106.3% in acetone for plasma, respectively. The calibration curves has shown good linearity with the correlation coefficients (R²) varying from 0.992 to 1.000 for urine and from 0.996 to 1.000 for plasma, respectively. The detection limits of all analytes were obtained in the range of 0.01-0.1 ng/mL for urine and 0.1-0.4 ng/mL for plasma, respectively. The metabolites of BaP were excreted as mono hydroxy and dihydrodiol forms after intraperitoneal injection of 20 mg/kg of BaP to rats. The total amounts of BaP and four metabolites excreted in dosed rat urine were 3.79 ng over the 0-96 hr period from adminstration and the excretional recovery was less than 0.065% of the injection amounts of BaP. The proposed method was successfully applied to the determination of BaP and its hydroxylated metabolites in rat urine and plasma for the pharmacokinetic studies.

Analysis of Peanut and Peanut Butter Retailed in Korea for Aflatoxin $B_1$ (국내 시판 땅콩 및 땅콩버터 중 Aflatoxin $B_1$ 오염 분석)

  • Park, Je-Won
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.309-312
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    • 2006
  • Aflatoxin $B_1$ in 70 retail samples, including 40 food-grade peanut (28 domestic, 12 imported) and 30 peanut butter (12 domestic, 18 imported) samples, was analyzed by high-performance liquid chromatography (HPLC) with fluorescence detection (FD), and positive samples were confirmed using HPLC with mass spectrometry (MS). Recoveries of aflatoxin $B_1$ spiked at 2 ppb exceeded 80% in both commodities. Detection limits for aflatoxin $B_1$ by HPLC-FD and MS analysis were 0.8 and 0.1 ppb, respectively. Four domestic and six imported peanut samples contained detectable levels of aflatoxin $B_1$ with means of 19 and 32 ppb, respectively. Aflatoxin $B_1$ was found in two domestic and three imported peanut butter samples with mean aflatoxin $B_1$ of 10 and 12 ppb, respectively. Peanut commodity showed more frequent aflatoxin $B_1$ contamination compared to its processed peanut butter product, and levels of aflatoxin $B_1$, especially in imported peanuts, were significantly (p<0.05) higher than those of other commodities. These results suggest peanut and peanut butter are not major contributors to dietary intake of aflatoxin $B_1$ in South Korea.

A Scientific Analysis of Gold Threads Used in Donggungbi-Wonsam(Ceremonial Robe Worn by a Crown Princess, National Folklore Cultural Heritage No.48) (동궁비 원삼에 사용된 금사의 과학적 분석)

  • Lee, Jang-Jon;An, Boyeon;Han, Kiok;Lee, Ryangmi;Yoo, Ji Hyun;Yu, Ji A
    • Journal of Conservation Science
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    • v.37 no.5
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    • pp.525-535
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    • 2021
  • This study identified material properties through scientific analysis on Jikgeumdan(satin with gold threads) from Donggungbi-Wonsam and the gold threads used in the embroidery. The Donggungbi-Wonsam's base of gold threads were estimated to have used mulberry fiber's Korean paper(Hanji) because non-wood-based fibers were observed. The X-ray spectrometer showed that the Tongsuseulan of Donggungbi-Wonsam was a flat gold thread of pure gold and Jikgeumdan of flat silver thread of its Saekdong and Hansam. High sulfur levels were detected in the flat silver thread, which appeared to have formed silver sulfide by either manufacturing process using sulfur or conservation environment. he dragon insignia's embroidery is also described as two types twisted gold threads; pure gold and alloying-gold and silver. while dragon insignia's border line is decorated with a twisted gold thread of pure gold. In particular, it was investigated that adhesives such as an animal glue, a protein-based compound by gas chromatography mass spectrometry. Additionally, XRF and Raman spectroscopy analysis on the mixture substances between the metal surface and the base paper of gold threads identified talc and quartz in the gold threads and Seokganju(hematite) in the flat silver threads.

A Study on the Quantitative Analysis of Portable XRF for the Components Analysis of Metal Cultural Heritage (금속문화재 성분분석을 위한 휴대용 XRF 정량분석법 연구)

  • Lim, So-Mang;Kwon, Young-Suk;Cho, Young-Rae;Chung, Won-Sub
    • Journal of Conservation Science
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    • v.37 no.5
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    • pp.451-463
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    • 2021
  • In this study we conducted component analyses of portable XRF detectors using four Au-Cu alloy standard samples to improve their accuracy by drawing up a calibration curve based on ICP-OES standard values. The portable XRF analysis found absolute errors of 0.3 to 3.7 wt% for Au and 0.2 to 8.2 wt% for Cu, confirming that the error range and standard deviation differed from one detector to another. Furthermore, the calibration curve improved their accuracy, such that the relative error rates of Au and Cu decreased from 9.8% and 14% to 3.5% and 3.7%, respectively. Accordingly, an experiment to confirm the calibration curve was conducted using unknown samples, finding that the measured values of the unknown samples fell on the calibration curve. Therefore, to accurately analyze the components of metal cultural heritage items, it is necessary to prepare a calibration curve for each element after checking whether the detector is suitable for the artifact.

Hydrothermal Alteration around the Tofua Arc (TA) 25 Seamounts in Tonga Arc (통가열도 TA 25 해저산의 열수변질)

  • Cho, Hyen Goo;Kim, Dong-Ho;Koo, Hyo Jin;Um, In Kwon;Choi, Hunsoo
    • Journal of the Mineralogical Society of Korea
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    • v.27 no.4
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    • pp.169-181
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    • 2014
  • Korea government has consistently investigated the development of economic mineral deposits in the Tofua volcanic arc, Tonga since 2008 for the secure of sea floor mineral resources. We studied the composition and distribution of minerals formed by hydrothermal activity around TA 25 seamounts of the Tofua volcanic arc, Lau Basin, Tonga, using X-ray diffraction analysis, scanning electron microscopy, X-ray fluorescence spectrometry, and inductively coupled plasma atomic emission spectrometry. We used 7 core samples and 9 surface sediment samples. Barite, sphalerite, and clinoclase are present in the most volcanic vent area. Gypsum, smectite, and kaolin mineral are distributed in vent A area, chalcopyrite, pyrite, smectite, and kaolin mineral are in vent B and C area, and gypsum, chalcopyrite, pyrite, and goethite are in vent D area. From the study of clay fraction, smectite and few kaolinite are detected in the most studied area except inner part of caldera, which suggest that argillic alteration are dominant in the volcanic vent areas. Various sulfide or arsenide minerals were found in the hydrothermal vent B, C, and D. The mineralogy and geochemistry suggest higher hydrothermal activities in volcanic vent B, C, and D compared to vent A and inner caldera area. Therefore higher probabilities of massive sulfide deposits may occur in hydrothermal vent B, C, and D.