Determination of Trace Germanium in Marine Sediments by Hydride Generation-Atomic Fluorescence Spectrometry （HG-AFS）

1　Introduction　Germaniumisatypicaldispersedelementintheearth’scrustwithanaveragecontentof 7mgkg- 1.Itisdistributedinthecrustintheformofsilicate ,sul fide ,sulfate (FroelichandAndreae ,1981)andsoon .Atpresent ,manymethods ,suchashydridegenera tion induc…     

Preliminary investigation of mercury in bone tissues of skua and penguin in Antarctica using AFS and Synchrotron Radiation X-ray Fluorescence （SR-XRF）

Mercury （Hg） was investigated in bone tissues of skua （ Catharacta maccormick） and penguin （Pygoscelis adeliae） collected in the maritime Antarctic using atomic fluorescence spectrometry （AFS） and synchrotron radiation X-ray fluorescence （SR-XRF） method. The total levels of mercury in bone tissues of penguin and skua are much lower than those in other organs （e. g. , kidney, liver）. The toxic effects of mercury in bone tissues of seabirds in polar region are not known. We have used SR- XRF method to map the distribution of trace levels of mercury in bones. The levels of mercury are found to be enriched somewhere near the periosteal surface and/or endosteal surface. The distribution of mercury shows strongly correlation with that of some essential elements and probably poses negative effect on the bone metabolism inferring from the relationship of mercury with the other elements. These studies represent a first step toward understanding the toxic effects of mercury on bone of polar animals by suggesting the possible microscopic investigation.     

Determination of Trace Copper （Ⅱ） in Water Samples by Kinetic-spectrophotometry

A new kinetic-spectrophotometric method is proposed for the determination of copper （ Ⅱ ）. The method is based on the catalytic effect of copper （ Ⅱ ） on the oxidation of weak acid brilliant blue dye （RAWL） by hydrogen peroxide. The copper （ Ⅱ ） can be determined spectrophotometrically by measuring the decrease of absorbance of RAWL at λ = 626 nm using the fix-time method. The optimum reaction conditions are as follows： pH 7.20, buffer solution NaOH-KH2PO4, RAWL （200 mgL-1） 5.00 mL, H2O2 （30%） 0.50 mL, reaction temperature 80 ？？ and reaction time 20 min. The linear range of this method is between 0 μg L^-1 and 12 μg L^-1 and the limit of detection is 0.011 μg L-1, the relative standard deviation （RSD） in five replicate determinations for 2 and 8 μg L-1 copper （ Ⅱ ） are 3.2% and 2.3%, respectively. Twenty ions do not interfere in the determination of copper （ Ⅱ ）. The method has been applied satisfactorily to the determination of copper （ Ⅱ ） in freshwater samples （tap water and Yellow River water from Lijin, Shandong, China） and seawater samples （from the South China Sea）, the recovery rates are 98.0%, 102.5% and 96.0%, respectively.     

Efficient expression of green fluorescent protein （GFP） mediated by a chimeric promoter in Chlamydomonas reinhardtii

To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein （GFP） was expressed in C. reinhardtii under the control of promoters： RBCS2 and HSP70A-RBCS2. Efficiency of transformation and expression were compared between two transgenic algae： RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ. In addition, a threefold increase of GFP in Tran-Ⅱ was induced by heat shock at 40℃. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii.     

Efficient expression of green fluorescent protein （GFP） mediated by a chimeric promoter in Chlamydomonas reinhardtii

To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii,a high efficient expression vector was constructed.Green fluorescent protein(GFP) was expressed in C.reinhardtii under the control of promoters:RBCS2 and HSP70A-RBCS2.Efficiency of transformation and expression were compared between two transgenic algae:RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ.Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2,and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ.In addition,a threefold increase of GFP in Tran-Ⅱwas induced by heat shock at 40°C.All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C.reinhardtii.     

Geochemical analysis of marine sediments using fused glass disc by X-ray fluorescence spectrometry

A method was developed for content determination of Na, Mg, A1, Si, P,S, C1, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Ga, Nb, Zr, Y, Sr, Rb, Ba, La and Ce etc. covering 26 major, minor, and trace elements in marine sediment samples using fused glass disc by X-ray Fluorescence spectrometry. Calibration was made using marine sediment certified reference materials and the synthetic standard samples prepared by mixing several marine sediments with stream sediment and carbonate standard samples in different proportions. The matrix effect was corrected using theoretical alpha coefficients, experience coefficients and the scattered radiation as the internal standard （for the trace elements）. The accuracy of the method was evaluated by analysis of certified reference materials GBW07314, GBW07334 and GSMS6. The results are in good agreement with the certified values of the standards with RSD less than 2.60%, except for Y, Cr, Ga, Ce, La, Nb, Rb, and V with RSD less than 9.0% （n=12）.     

Distribution and Sources of Polycyclic Aromatic Hydrocarbon （PAH） in Marine Environment of China

1INTRODUCTIONPolycyclicaromatichydrocarbons (PAHs)depo sit edinmarineenvironmentsfromavarietyofsourcessuchaswastewater,industrialanddomesticdischarges,andoilspills.Themajorityofthesesourcesfrompetrogenicoriginischaracterizedbyadominanceoflowmolecularweightaromaticcompounds,especiallynaphthaleneandalkylatedPAHs(Sportoletal.,1 983 ) .Crudeoilanditsrefinedproductssuchasgasoline,keroseneandotherfu eloilshaveahighcontentofalkylatedPAHsduetotheirslowformationattemperatureslowerthanthatofcomb…     

Monte carlo simulation of in situ gamma-spectra recorded by NaI (Tl) detector in the marine environment

To develop a Na I(Tl) detector for in situ radioactivity monitoring in the marine environment and enhance the confidence of the probability of the gamma-spectrum analysis, Monte Carlo simulations using the Monte Carlo N-Particle( MNCP) code were performed to provide the response spectra of some interested radionuclides and the background spectra originating from the natural radionuclides in seawater recorded by a Na I(Tl) detector. A newly developed 75 mm × 75 mm Na I(Tl) detector was calibrated using four reference radioactive sources 137 Cs, 60 Co, 40 K and 54 Mn in the laboratory before the field measurements in seawater. A simulation model was established for the detector immersed in seawater. The simulated spectra were all broadened with Gaussian pulses to reflect the statistical fluctuations and electrical noise in the real measurement. The simulated spectra show that the single-energy photons into the detector are mostly scattering low-energy photons and the high background in the low energy region mainly originates from the Compton effect of the high energy γ-rays of natural radionuclides in seawater. The simulated background spectrum was compared with the experimental one recorded in field measurement and they seem to be in good agreement. The simulation method and spectra can be used for the accurate analysis of the filed measurement results of low concentration radioactivity in seawater.     

Determination of trace vanadium (V) in seawater and fresh water by the catalytic kinetic spectrophotometric method

A new kinetic spectrophotometric method has been developed for the determination of vanadium (V). The method is based on the catalytic effect of vanadium (V) on the oxidation of weak acid brilliant blue dye (RAWL) by KBrO₃ using the citric acid as activation reagent. The obtained optimum conditions are: c (RAWL) = 1×10⁻⁴ molL⁻¹, c (KBrO₃) = 3×10⁻² molL⁻¹, c (citric acid) = 9×10⁻³ molL⁻¹, pH = 2.50, the reaction time being 7.0 min and the temperature being 25.0°C. Under the optimum conditions, the proposed method allows the determination of vanadium (V) in the range of 0–70.0 ng mL⁻¹ and the detection limit is down to 0.407 ng mL⁻¹. For standard vanadium (V) solution determination, the recovery efficiency is in the range of 98.5%–102% and the RSD ranges from 0.76%–1.25%. Moreover, it is demonstrated that most cations and anions do not interfere with the determination of vanadium (V) under the analytical condition. The new method was successfully applied in the determination of vanadium (V) in fresh water and seawater samples with satisfactory results. Vanadium (V) in the seawater samples from Qingdao offshore was determined using the method and the distribution of vanadium (V) was mapped. Compared with other instrumental analytical methods, the proposed method shows good selectivity, sensitivity, simplicity, lower cost and rapidity. It can be employed on shipboard easily.     

Determination of Ca,K, Mg and Fe in four fish species by FAAS

A method for determining calcium(Ca),potassium(K),magnesium(Mg) and iron(Fe) in four fish species was optimized and validated.It included microwave mineralization of the samples and subsequent quantification by flame atomic absorption spectroscopy(FAAS) with Zeeman-effect background correction.Using HNO3(65%) and H2O2(33%) as extraction solutions,the optimal conditions of extraction were established as follows:0.5 g of sample mass;microwave time program of 300 W/5 min and 600 W/5 min.The method was free of matrix interferences.The linear correlation coefficients were ≥0.9991,the recovery percentage of analytes was from 99.31% to 103.70% and the RSD(relative standard deviation) was lower than 2.06%.The detection limits obtained were 32.3,43.2,14.0 and 68.6 ng mL-1 for Ca,K,Mg and Fe in FAAS respectively.It is shown that the method is rapid,simple,sensitive and accurate.The method was applied to the studies of digestibility and measurement of these nutrients in samples of fish collected from Norway,Japan and China.     

Optimization of fluorescence in situ hybridization （FISH） for the identification of two polar coccoid green algae species

Standard FISH protocols using fluorochrome-labeled oligonucteotide probes have been successfully applied for in situ detection.However,optimized protocols of FISH for specific eukaryotes in marine environments are often not developed.This study optimized the conditions of fluorescence in situ hybridization (FISH) by using two polar isolated microalgae.The modified conditions were as follows: (1) 10 mg·mL-1 lysozyme solution pretreatment at 37℃ for 30 min;(2) the hybridization buffer including 20% formamide;(3) the hybridization condition was 47℃ for 6 h. The cells enumerated by FISH were compared with those enumerated by flow eytometry (FCM) and DAPI to confirm the cell loss and hybridization efficiency.The optimized protocol was also successfully applied to Arctic Ocean samples,which were found to be dominated by Micromonas sp.The modified protocol showed a high relative efficiency and could be successfully applied for the detection of specific microbial eukaryotes in environmental samples.     

Identification and characterization of a new IgE-binding protein in mackerel (Scomber japonicus) by MALDI-TOF-MS

As fish is one source of the ‘big eight’ food allergens,the prevalence of fish allergy has increased over the past few years.In order to better understand fish allergy,it is necessary to identify fish allergens.Based on the sera from fish-allergenic patients,a 28 kDa protein from local mackerel (Scomber japonicus),which has not been reported as a fish allergen,was found to be reactive with most of the patients’ sera.The 28 kDa protein was analyzed by MALDI-TOF-MS (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry).Mascot search in NCBI database (Date: 08/07/2010) showed that the top protein matched,i.e.triosephosphate isomerase (TPI) from Xiphophorus maculatus and Poecilia reticulata,had a mowse (molecular weight search) score of 98.In addition,TPI from Epinephelus coioides also matched this mackerel protein with a mowse score of 96.Because TPI is con-sidered as an allergen in other non-fish organisms,such as lychee,wheat,latex,archaeopotamobius (Archaeopotamobius sibiriensis) and crangon (Crangon crangon),we consider that it may also be an allergen in mackerel.