Determination of tetrodotoxin by liquid chromatography coupled with electrospray ion-trap mass spectrometry

Two methods for tetrodotoxin analysis using liquid chromatography coupled with electrospray iontrap mass spectrometry (LC-ESI-MS) have been established with C,, reversed phase column and hydrophilic interaction liquid chromatography (HILIC) column, respectively. Sensitivity and reproducibility of the methods were compared. The method using C-18 column in selected ion monitoring (SIM) mode had a detection limit (S/N = 3) of 120 pg, and a good linearity of the calibration curve was obtained for tetrodotoxin (r = 0. 9992). High reproducibility of the method was observed, with a relative standard deviation (RSD) below 10%. The method using HILIC column in SIM mode and selected reaction monitoring (SRM) mode had detection limits (S/N = 3) of 15 and 3.75 pg, respectively. Good linearity of the calibration curves was obtained for tetrodotoxin (r = 0. 9996 and 0. 9998 in SIM and SRM mode, respectively). T he reproducibility was high in SIM mode but relatively poor in SRM mode. Based on the results, the method using HILIC column in SIM mode was suggested for the analysis of tetrodotoxin with LC-MS system.

A three-dimensional coupled physical-biological model study in the spring of 1993 in the Bohai Sea of China

A three-dimensional (3-D) coupled physical and biological model was used to investigate the physical processes and their influence on the ecosystem dynamics of the Bohai Sea of China. The physical processes include M-2 tide, time - varying wind forcing and river discharge. Wind records from I to 31 May in 1993 were selected to force the model. The biological model is based on a simple, nitrate and phosphate limited, lower trophic food web system. The simulated results showed that variation of residual currents forced by M, tide, river discharge and time-varying wind had great impact on the distribution of phytoplankton biomass in the Laizhou Bay. High phytoplankton biomass appeared in the upwelling region. Numerical experiments based on the barotropic model and baroclinic model with no wind and water discharge were also conducted. Differences in the results by the baroclinic model and the barotropic model were significant: more patches appeared in the baroclinic model comparing with the barotropic model. And in the baroclinic model, the subsurface maximum phytoplankton biomass patches formed in the stratified water.

Bromophenols coupled with methyl gamma-ureidobutyrate and bromophenol sulfates from the red alga Rhodomela confervoides

Four new bromophenols C-N coupled with methyl gamma-ureidobutyrate (1-4), a phenylethanol bromophenol (5), and three phenylethanol sulfate bromophenols (6-8) have been isolated from polar fractions of an ethanolic extract of the red alga Rhodomela confervoides. On the basis of spectroscopic evidence including HRMS and 2D NMR data, the structures of the new compounds were determined as methyl N'-(2,3-dibromo-4,5-dihydroxybenzyl)-gamma-ureidobutyrate (1), methyl N,N'-bis(2,3-dibromo-4,5-dihydroxybenzyl)-gamma-ureidobutyrate (2), methyl N'-[3-bromo-2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxybenzyl]-gamma-ureidobutyrate (3), methyl N'-(2,3-dibromo-4,5-dihydroxybenzyl)-A7-[3-bromo2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxybenzyl]-gamma-ureidobutyrate (4), 2,3-dibromo-4,5-dihydroxyphenylethanol (5), 2,3-dibromo-4,5-dihydroxyphenylethanol Sulfate (6), 3-bromo-4,5-dihydroxyphenylethanol sulfate (7), and 3-bromo2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxyphenylethanol sulfate (8). The cytotoxicity of all compounds was evaluated against several human cancer cell lines including human colon cancer (HCT-8), hepatoma (Bel7402), stomach cancer (BGC-823), lung adenocarcinoma (A549), and human ovarian cancer (A2780). Among them, the phenylethanol and the phenylethanol sulfate bromophenols (5-8) showed moderate cytotoxicity against all tested cell lines.

Bromophenols coupled with nucleoside bases and brominated tetrahydroisoquinolines from the red alga Rhodomela confervoides

Three new bromophenols C-N coupled with nucleoside base derivatives (1-3) and three new brominated 1,2,3,4-tetrahydroisoquinolines (5-7, together with a new brominated tyrosine derivative (4, have been isolated from polar fractions of an ethanolic extract of the red alga Rhodomela confervoides. By spectroscopic and chemical methods including HRMS and 2D NMR data, their structures were determined as 7-[3-bromo-2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxybenzyl]-3,7-dihydro-1H-purine-2,6-dione (1), 7-(2,3-dibromo-4,5-dihydroxybenzyl)-3,7-dihydro-1H-purine-2,6-dione (2, 9-[3-bromo-2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxybenzyl]adenine (3), (-)-8S-(3-bromo-5-hydroxy-4-methoxy)phenylalanine (4), (-)-3S-8-bromo-6-hydroxy-7-methoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (5), methyl (-)-3S-8-bromo-6-hydroxy-7-methoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylate (6), and methyl (-)-3S-6-bromo-8-hydroxy-7-methoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylate (7). Compounds 5-7 were semisynthesized by using 4 as the starting material.

Bromophenols coupled with derivatives of amino acids and nucleosides from the red alga Rhodomela confervoides

Three new bromophenols coupled with pyroglutamic acid derivatives and one bromophenol coupled with deoxyguanosine were obtained from the red alga Rhodomela confervoides. By spectroscopic methods including 2D NMR and single-crystal X-ray structure analysis their structures were elucidated as N-(2,3-dibromo-4,5-dihydroxybenzyl)methyl pyroglutamate (1), N-(2,3-dibromo-4,5-dihydroxybenzyl)pyroglutamic acid (2), N-[3-bromo-2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxybenzyllmethyl pyroglutamate (3), and 2-N-(2,3-dibromo-4,5-dihydroxybenzylamino)deoxyguanosine (4), respectively. Compounds 1-4 were evaluated against several microorganisms and human cancer cell lines, but found inactive. To our knowledge this is the first report of bromophenols coupled with amino acid or nucleoside derivatives through the C-N bond.

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.

Speciation Analysis of Organotin Compounds in Shellfish by Hyphenated Technique of 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 in the shellfish samples. Agilent TC-C-18 column was selected, mobile phase of the HPLC was CH3CN:H2O: CH3COOH = 65:23:12 (V/V), 0. 05% TEA, pH = 3.0 at flow rate 0.4 mL/min. Five mixed organotin standards from 100 mu g/L to 0. 5 mu g/L was used for the method evaluation. The experimental results indicate that the linearity (R-2) for each compound was over 0.998. The shellfish samples were treated by supersonic extraction with mobile phase for 30min. Four organotin compounds including dibutyltin (DBT), tributyltin (TBT), diphenyltin (DphT) and triphenyltin (TPhT) in shellfish samples were detected with method mentioned above. It was found that the domain compounds in the samples were tributyltin (TBT) and triphenyltin (TPhT). The recoveries test from the standard addition for trimethyltin (TMT tributyltin (TBT), and triphenyltin (TPhT) were, over 80%. However, the recoveries for diphenyltin (DPhT) and dibutyltin (DBT) were relatively low, 37.3% and 75.2% respectively. The reason might be attributed to the decomposition of those compounds during the extraction procedure. The further study on this subject is under the progress.

Fingerprinting quality control of Qianghuo by high-performance liquid chromatography-photodiode array detection

A novel, simple and accurate fingerprint method was developed using high-performance liquid chromatography-photodiode array detection (HPLC-DAD) for the quality control of Qianghuo, a Tibetan folk and Chinese herbal medicine used as a diaphoretic, an antifebrile and an anodyne. For the first time, the feasibility and advantages of employing chromatographic fingerprint were investigated and demonstrated for the evaluation of Qianghuo by systematically comparing chromatograms of aqueous extracts with the professional analytical software recommended by State Food and Drug Administration (SFDA). Our results revealed that the chromatographic fingerprint combing similarity evaluation could efficiently identify and distinguish raw herbs of Qianghno from different sources and different species. The effects on Notopterygium forbesii Boiss (Apiaceae) chromatographic fingerprints resulted from collecting locations, harvesting time were also examined. (c) 2006 Elsevier lrelanc Ltd. All rights reserved.

泥河湾盆地磁性地层学研究——以东谷索和岑家湾剖面为例

The Nihewan Basin is a key area for research into human occupation at high northern latitudes in northeast Asia after the initial expansion of early humans out of Africa. Well-developed late Cenozoic lacustrine deposit sequence in this basin offers a unique opportunity to address this issue. In this thesis, detailed magnetostratigraphic investigation coupled with mineral magnetism was conducted on the Donggutuo and Cenjiawan sections in the eastern basin, where lacustrine deposits sequences containing the Donggutuo, Maliang and Cenjiawan Paleolithic sites are well developed. The sequences are mainly composed of grayish-white clays, grayish-green clayey silts, grayish-yellow silts and fine-grained brown sands, which have recorded reliable polarity variations of geomagnetic field.Characteristics of the anisotropy of magnetic susceptibility show that the sediments have preserved typical original magnetic fabric for sediments, indicating that the strata were developed in a low-energy lake environment and were never perturbed by tectonic stress since deposition. High-temperature magnetic susceptibility measurements (x-T) of representative specimens and demagnetization experiments indicate that the dominant magnetic mineral and remanent carrier at the two sections is magnetite. In addition, hematite and possibly minor maghemite exists in some portions of the sequences. The majority of the samples have relatively simple demagnetization behaviors. After removal of soft magnetic components, the stable characteristic remanent magnetizations (ChRMs) are isolated, which can represent the original remanences.The Donggutuo section mainly records the Brunhes chron, the Matuyama chron and the Jaramillo subchron. The Maliang stone artifact layer occurs just below the Brunhes/Matuyama boundary; and the Donggutuo artifact layer, just below the Jaramillo onset. Accordingly, the Maliang and Donggutuo sites can be dated at about 0.78 Ma and 1.1 Ma, respectively. The Cenjiawan section has recorded a portion of the Matuyama chron. After correlations with the magnetic polarity sequence of the Majuangou section adjacent to this section, the Cenjiawan stone artifact layer is determined below the Jaramillo onset, with an estimated age of 1.1 Ma.To establish the magnetic stratigraphy framework for the lacustrine sediments in the eastern Nihewan Basin, this thesis draws on the magnetic polarity sequences of the Donggutuo and Cenjiawan sections as well as previously obtained results from the Majuangou, Haojiatai, Xiaochangliang and Donggou sections for magnetostratigraphic correlations. The accumulation of the lacustrine sequences at the east margin of the basin commenced from about 2.0 Ma. These sequences record not only the coarse magnetostratigraphy of the Brunhes normal chron and the middle to late Matuyama reverse chron (that is, the Jaramillo and Olduvai subchrons) but also some of the fine structure (that is, the Kamikatsura, Santa Rosa, Punaruu and Cobb Mountain geomagnetic events). The development of the Nihewan paleolake experienced at least twice large expansion periods, split by a large-scale shrinking event in the middle period of the paleolake development. The accumulation of the lacustrine strata was controlled by fault activities.After temporal control for the Donggutuo, Maliang and Cenjiawan Paleolithic sites were established, the three sites along with other well-dated Paleolithic/hominin sites of the Early Pleistocene in North China were combined to construct a chronological sequence of early human occupation in northeastern Asia. Furthermore, after incorporation of paleoclimate changes retrieved from Chinese loess/paleosol sequences and marine sediments, it could be possibly proposed that human groups of the Early Pleistocene in North China might have survived repeated warm/humid interglacials and cold/dry glacials, which were paced by earth orbital variations of the Eastern paleomonsoon.