A simple method for DNA extraction from sporophyte in the brown alga Laminaria japonica

A simple method was developed for extracting DNA from brown algae Laminaria japonica, which possess large amounts of acidic polysaccharides. Firstly, the sporophyte were washed by eliminating polysaccaride buffer to remove the polysaccharides and then ground in liquid nitrogen. Secondly, the powders were treated with lysing buffer. Thirdly, KAc was used to eliminate the remaining acidic polysaccharides. The extracted DNA was purified using a chloroform-isoamyl alcohol ( 24: 1 v/v), and precipitated in cold isopropanol. The yield was from 18.7 to 37.5 mu g g(-1) (wet weight) and the purity of total DNA was determined spectrophotometrically as the ratio of A(260)/A(280), which was about 1.7 - 1.9. The extracted DNA was of high quality and suitable for molecular analyses, such as PCR, restriction enzyme digestion. This method is a reproducible, simple, and rapid technique for routine DNA extraction from sporophyte in Laminaria japonica. Furthermore, the low cost of this method makes it attractive for large-scale studies.

Extraction of phosphorus in natural waters and sediments by ignition method

Phosphorus recoveries from selected inorganic and organic P-containing compounds after ignition with auxiliaries, such as MgSO4, Mg (NO3)(2), MgCl2, Mg (Ac)(2) and CaCl2 were studied. It was found that the phosphorus could not be completely recovered when most P-containing compounds were ignited with MgSO4 at temperature not higher than 500degreesC if the baked residue was extracted with 0.2 mol/L HCl for 30 min at 80degreesC or at room temperature. In contrast, MgCl2, Mg(Ac)(2) and CaCl2, as well as Mg(NO3)(2), could all yield complete P recoveries. We suggest that MgCl2 rather than MgSO4, which is usually used, should be utilized as ashing auxiliary agent in the P extraction by ignition method. Although Mg (NO3)(2) is a highly effective auxiliary agent, yet interference from MgSO4, danger of explosion, toxicity of nitrogen dioxide and more manipulation steps may limit its widespread utilization. It is suggested that if sediment is ignited with MgCl2, the extraction of residue with 0.2 mol/L HCl for 30 min at 80degreesC could give good result.

An efficient combination of supercritical fluid extraction and high-speed counter-current chromatography to extract and purify homoisoflavonoids from Ophiopogon japonicus (Thunb.) Ker-Gawler

Supercritical fluid extraction (SFE) was used to extract homoisoflavonoids from Ophiopogon japonicus (Thunb.) Ker-Gawler. The optimization of parameters was carried out using an orthogonal test L-9 (3)(4) including pressure, temperature, dynamic extraction time and the amount of modifier. The process was then scaled up by 100 times with a preparative SFE system under the optimized conditions of 25 MPa, 55 degrees C, 4.0 h and 25% methanol as a modifier. Then crude extracts were separated and purified by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane/ethyl acetate/methanol/ACN/water (1.8:1.0:1.0:1.2:1.0 v/v). There three homoisoflavonoidal compounds including methylophiopogonanone A 6-aldehydo-isoophiopogonone A, and 6-formyl-isoophiopogonanone A, were successfully isolated and purified in one step. The collected fractions were analyzed by HPLC. In each operation, 140 mg crude extracts was separated and yielded 15.3 mg of methylophiopogonanone A (96.9% purity), 4.1 mg of 6-aldehydo-isoophiopogonone A (98.3% purity) and 13.5 mg of 6-formyl-isoophiopogonanone A (97.3% purity) respectively. The chemical structure of the three homoisoflavonoids are identified by means of ESI-MS and NMR analysis.

Study on extraction of agaropectin from Gelidium amansii and its anticoagulant activity

Gelidium amansii agar was fractionated on DEAE-cellulose and four fractions were obtained sequentially. The yields of 1.0 mol/L NaCl fraction and 2.5 mol/L NaCl fraction were 2.80% and 2.03%. They are highly sulfated agar, and named as agaropectin with sulfate content being 22.8% and 32.5%, respectively. The anticoagulant experiment results show that agaropectin could effectively prolong the coagulation time in a dose-dependent manner in vitro. Agaropection could be absorbed and effectively prolong the plasma coagulation time in vivo. After intragastric administration at the doses of 100, 200, and 400 mg/kg.d in rats for 15 days, TT (thrombin time), CT (coagulation time), PT (prothrombin time), and APTT (activated partial thromboplastin time) could be effectively prolonged and the plasma Fib level could be significantly lowered.

The extraction of pigments from fresh Laminaria japonica

The pigments in Laminaria japonica was extracted with six organic solvents and analyzed in spectroscopy analysis. The extractions conditions were screened by an orthogonal test and the quantity of extracted pigments was determined spectroscopically. The results show that: (1) among the six organic solvents, acetone was the most effective one for the extraction; (2) the optimum extraction conditions were as follows: the ratio of S/M (solvent volume/ material weight) was 30 ml/g; fresh seaweed was extracted 2 times in 2 h; (3) the average total content of pigments was 1.85 mg/g (calculated with dry L. japonica).

Extraction of the polysaccharides from five algae and their potential antioxidant activity in vitro

The sulfated polysaccharides extracted from algae possess excellent antioxidant activities. In this study, we prepared five polysaccharides extracted from five algae including one brown alga Laminaria japonica. one red alga Porphyra haitanensis and three green algae Ulva pertusa, Enteromorpha linza and Bryopsis plumose. And then the antioxidant activities of all the samples were investigated including scavenging effects of superoxide and hydroxyl radicals, and reducing power. The chemical analysis and FT-IR spectrum showed these extracts were polysaccharides. And in addition, we found that certain polysaccharide exhibited stronger antioxidant activity in certain antioxidant activity. Factors effecting and attributing to radical scavenging effect need to be further studied. (C) 2010 Elsevier Ltd. All rights reserved.

Simultaneous Determination of Multi-Organotin Compounds in Seawater by Liquid-Liquid Extraction-High Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry

The hyphenated technique of high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (HPLC-ICP-MS) was applied to the simultaneous determination of five organotin compounds (trimethyltin, dibutyltin, tributyltin, diphenyltin and triphenyltin) in seawater samples. Agilent TC-C18 column was used for the separation, the mobile phase of HPLC was CH3CN : H2O: CH3COOH=65 : 23 : 12 (phi), 0.05% TEA, and pH value was adjusted to 3.0 by diluent ammonia. The flow rate was 0.6 mL . min(-1). Five mixed organotin compounds in a mix standard solution from 100 to 0.5 mu g . L-1 were applied for the method assessment. The experimental results indicate that the correlation coefficient of calibration curves (R-2) for each organotin compound was over 0.998 and the detection limits of the five organotin compounds were lower than 3 ng . L-1. Different mixed organic solvents including dichloromethane or toluene were used for extraction of organotin and the extraction condition of organotin from seawater was optimized. The 100 mL seawater acidized by hydrochloric acid was extracted by 10 mL carbon dichloride (CH2Cl2) with 2% tropolone for 10 min twice. Extracted organic solvents were mixed And blown to one drop by nitrogen with the rate of 1.7 mL . min(-1), then 1 mL acetonitrile was added to the drop for redissolving the organotin compounds. Finally, the mixed redissolution was filtered by 0.22 mu m organic filter membrane before analysis. it was found that the only organotin compound in seawater was triphenyltin (TPHT) and the content was 53.2 ng . L-1. The recoveries test from the standard addition for diphenyltin (DPHT), dibutyltin (DBT), tributyltin (TBT) and triphenyltin (TPHT) were over 80%. However, the recovery for trimethyltin (TMT) was relatively low and the value was 50%. The reason might be attributed to the decomposition or adsorption of those compounds during the extraction procedure. Further study on this subject is in progress.

The study of immersion chloride extraction of iron artifacts

The chloride extraction rules of iron artifacts were studied by immersion methods. Different chloride extraction results between the alkaline solution and a washing solution were obtained. The microstructure and the anti-corrosion performance of the samples before and after treatment, were, respectively studied by the scanning electron microscope (SEM) and the potentiodynamic scanning method. The results indicated that Cl- removed according to the diffusion law. The microstructure of the samples transformed after treatment. The rusts became more compact, and the porosity also increased. The chloride extraction effect in the washing solution was better than that in the NaOH solution.

A study of electrical chloride extraction of iron artifacts

The chloride extraction rule of iron artifacts was studied by electrical methods. The effect of the current and potential value on the desalination result of simulated iron artifacts was studied through the galvanostatic and potentiostatic experiments the ingredients of the rust before and after treatments were also analyzed by the X-ray diffraction (XRD). It has been found that the optimal current density was between -0.50 and -0.75 mA/cm(2) and the optimal potential was between -1.175 and -1.200 V. The phase of the samples rusts transformed after treatment, as well as the anti-corrosion performance improved.

Extraction of Nitraria tangutorum seed oil by supercritical carbon dioxide and determination of free fatty acids by HPLC/APC/IMS with fluorescence detection

The seed oil from Nitraria tangutorum samples was obtained by supercritical carbon dioxide extraction methods. The extraction parameters for this methodology, including pressure, temperature, particle size and extraction time, were optimized. The free fatty acids in the seed oil were separated with a pre-column derivation method and 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-p-toluenesulfonate (BDETS) as a labeling regent, followed by high-performance liquid chromatography (HPLC) with fluorescence detection. The target compounds were identified by mass spectrometry with atmospheric pressure chemical ionization (APCI in positive-ion mode). HPLC analysis shows that the main compositions of the seed oil samples were free fatty acids (FFAs) in high to low concentrations as follows: linoleic acid, oleic acid, hexadecanoic acid and octadecanoic acid. The assay detection limits (at signal-to-noise of 3:1) were 3.378-6.572 nmol/L. Excellent linear responses were observed, with correlation coefficients greater than 0.999. The facile BDETS derivatization coupled with mass spectrometry detection allowed the development of a highly sensitive method for analyzing free fatty acids in seed oil by supercritical CO2 extraction. The established method is highly efficient for seed oil extraction and extremely sensitive for fatty acid profile determination. (C) 2007 Elsevier B.V. All rights reserved.

胜利勘探数据中心及勘探辅助决策系统研究

In the paper through extensive study and design, the technical plan for establishing the exploration database center is made to combine imported and self developed techniques. By research and repeated experiment a modern database center has been set up with its hardware and network having advanced performance, its system well configured, its data store and management complete, and its data support being fast and direct. Through study on the theory, method and model of decision an exploration decision assistant schema is designed with one decision plan of well location decision support system being evaluated and put into action. 1. Study on the establishment of Shengli exploration database center Research is made on the hardware configuration of the database center including its workstations and all connected hardware and system. The hardware of the database center is formed by connecting workstations, microcomputer workstations, disk arrays, and those equipments used for seismic processing and interpretation. Research on the data store and management includes the analysis of the contents to be managed, data flow, data standard, data QC, data backup and restore policy, optimization of database system. A reasonable data management regulation and workflow is made and the scientific exploration data management system is created. Data load is done by working out a schedule firstly and at last 200 more projects of seismic surveys has been loaded amount to 25TB. 2. Exploration work support system and its application Seismic data processing system support has the following features, automatic extraction of seismic attributes, GIS navigation, data order, extraction of any sized data cube, pseudo huge capacity disk array, standard output exchange format etc. The prestack data can be accessed by the processing system or data can be transferred to other processing system through standard exchange format. For supporting seismic interpretation system the following features exist such as auto scan and store of interpretation result, internal data quality control etc. the interpretation system is connected directly with database center to get real time support of seismic data, formation data and well data. Comprehensive geological study support is done through intranet with the ability to query or display data graphically on the navigation system under some geological constraints. Production management support system is mainly used to collect, analyze and display production data with its core technology on the controlled data collection and creation of multiple standard forms. 3. exploration decision support system design By classification of workflow and data flow of all the exploration stages and study on decision theory and method, target of each decision step, decision model and requirement, three concept models has been formed for the Shengli exploration decision support system including the exploration distribution support system, the well location support system and production management support system. the well location decision support system has passed evaluation and been put into action. 4. Technical advance Hardware and software match with high performance for the database center. By combining parallel computer system, database server, huge capacity ATL, disk array, network and firewall together to create the first exploration database center in China with reasonable configuration, high performance and able to manage the whole data sets of exploration. Huge exploration data management technology is formed where exploration data standards and management regulations are made to guarantee data quality, safety and security. Multifunction query and support system for comprehensive exploration information support. It includes support system for geological study, seismic processing and interpretation and production management. In the system a lot of new database and computer technology have been used to provide real time information support for exploration work. Finally is the design of Shengli exploration decision support system. 5. Application and benefit Data storage has reached the amount of 25TB with thousand of users in Shengli oil field to access data to improve work efficiency multiple times. The technology has also been applied by many other units of SINOPEC. Its application of providing data to a project named Exploration achievements and Evaluation of Favorable Targets in Hekou Area shortened the data preparation period from 30 days to 2 days, enriching data abundance 15 percent and getting information support from the database center perfectly. Its application to provide former processed result for a project named Pre-stack depth migration in Guxi fracture zone reduced the amount of repeated process and shortened work period of one month and improved processing precision and quality, saving capital investment of data processing of 30 million yuan. It application by providing project database automatically in project named Geological and seismic study of southern slope zone of Dongying Sag shortened data preparation time so that researchers have more time to do research, thus to improve interpretation precision and quality.

Analysis of urinary 8-hydroxydeoxyguanosine by capillary electrophoresis and solid-phase extraction

Urinary 8-hydroxydeoxyguanosine (80HdG) has been considered as an excellent marker of individuals at high risk of developing cancer. Until now, urinary 80HdG has largely been measured by high-performance liquid chromatography with electrochemical detection. A new method for the analysis of urinary 80HdG by high-performance capillary electrophoresis has been developed and optimized in our laboratory. A single step solid-phase extraction procedure was optimized and used for extracting 80HdG from human urine. Separations were performed in an uncoated silica capillary (50 cm x 50 tm i.d.) using a P/ACE MDQ system with UV detection. The separation of 80HdG from interfering urinary matrix components is optimized with regard to pH, applied voltage, pressure injection time and concentration of SDS in running buffer. The detection limit of this method is 0.4 mug/ml, the linear range is 0.8-500 mug/ml, the correlation coefficients levels is better than 0.999. The developed method is simple, fast and good reproducibility, furthermore, it requires a very small injection volumes and low costs of analysis, which makes it possible to provide a new noninvasive assay for an indirect measurement of oxidative DNA damage.