Determination of free fatty acids in soil and bryophyte plants by precolumn derivatization via HPLC with fluorescence detection and tandem mass spectrometry (HPLC-MS/MS)

A sensitive method for the determination of 30 kinds of free fatty acids (FFAs, C-1-C-30) with 1-[2-(p-toluenesulfonate)-ethyl]-2-phenylimidazole-[4,5-f] 9,10-phenan- threne (TSPP) as labeling reagent and using high performance liquid chromatography with fluorescence detection and identification by online postcolumn mass spectrometry with atmospheric pressure chemical ionization (APCI) source in positive-ion mode (HPLC/MS/APCI) has been developed. TSPP could easily and quickly label FFAs in the presence of K2CO3 catalyst at 90 degrees C for 30 min in N,N-dimethylformamide (DMF) solvent, and maximal labeling yields close to 100% were observed with a 5-fold excess of molar reagent. Derivatives were stable enough to be efficiently analyzed by high performance liquid chromatography. TSPP was introduced into fatty acid molecules and effectively augmented MS ionization of fatty acid derivatives and led to regular MS and MS/MS information. The collision induced cleavage of protonated molecular ions formed specific fragment ions at m/z [MH](+)(molecular ion), m/z [M'+CH2CH2](+)(M' was molecular mass of the corresponding FFA) and m/z 295.0 (the, mass of protonated molecular core structure of TSPP). Fatty acid derivatives were separated on a reversed-phase Eclipse XDB-C-8 column (4.6 x 150 mm, 5 mu m, Agilent) with a good baseline resolution in combination with a gradient elution. Linear ranges of 30 FFAs are 2.441 x 10(-3) to 20 mu mol/L, detection limits are 3.24 similar to 36.97 fmol (injection volume 10 mu L, at a signal-to-noise ratio of 3, S/N 3:1). The mean interday precision ranged from 93.4 to 106.2% with the largest mean coefficients of variation (R.S.D.) < 7,5%. The mean intraday precision for all standards was < 6.4% of the expected concentration. Excellent linear responses were observed with correlation coefficients of > 0.9991. Good compositional data could be obtained from the analysis of extracted fatty acids from as little as 200 mg of bryophyte plant samples.Therefore, the facile TSPP derivatization coupled with HPLC/MS/APCI analysis allowed the development of a highly sensitive method for the quantitation of trace levels of short and long chain fatty acids from biological and natural environmental samples.

Determination of long-chain fatty acids in bryophyte plants extracts by HPLC with fluorescence detection and identification with MS

A sensitive method for the determination of long-chain fatty acids (LCFAs) (>C20) using 1-[2-(p-toluenesulfonate)-ethyl]-2-phenylimidazole-[4.5-f]-9,10-phenanthrene (TSPP) as tagging reagent with fluorescence detection and identification with post-column APCI/MS has been developed. The LCFAs in bryophyte plant samples were obtained based on distillation extraction with 1: 1 (v/v) chloroform/methanol as extracting solvent. TSPP could easily and quickly label LCFAs at 90 degrees C in the presence of K2CO3 catalyst in DMF. Eleven free LCFAs from the extracts of bryophyte plants were sensitively determined. Maximal labeling yields close to 100% were observed with a five-fold excess of molar reagent. Separation of the derivatized fatty acids exhibited a good baseline resolution in combination with a gradient elution on a reversed-phase Eclipse XDB-C-8 column. Calculated detection limits from 1.0 pmol injection, at a signal-to-noise ratio of 3, were 26.19-76.67 fmol. Excellent linear responses were observed with coefficients of >0.9996. Good compositional data were obtained from the analysis of the extracted LCFAs containing as little as 0.2 g of bryophyte plant samples. Therefore, the facile TSPP derivatization coupled with HPLC/APCI/MS analysis allowed the development of a highly sensitive method for the quantitation of trace levels of LCFAs from biological and natural environmental samples. (c) 2006 Elsevier B.V. All rights reserved.

Analysis of saturated free fatty acids from pollen by HPLC with fluorescence detection

A sensitive method for the determination of free fatty acids using 2-(2-(anthracen-10-yl)-1H-naphtho[2,3-dimidazol-1-yl) ethyl-p-toluenesuIfonate (ANITS) as tagging reagent with fluorescence detection has been developed. ANITS could easily and quickly label fatty acids in the presence of the K2CO3 catalyst at 90 degrees C for 40 min in N,N-dimethylformamide solvent. From the extracts of rape bee pollen samples, 20 free fatty acids were sensitively determined. Fatty acid derivatives were separated on a reversed-phase Eclipse XDB-C8 column by HPLC in conjunction with gradient elution. The corresponding derivatives were identified by post-column APCI/MS in positive-ion detection mode. ANITS-fatty acid derivatives gave an intense molecular ion peak at mlz [M+H](+); with MS/MS analysis, the collision-induced dissociation spectra of m/z [M+H](+) produced the specific fragment ions at mlz [M-345](+) and mlz 345.0 (here, m/z 345 is the core structural moiety of the ANITS molecule). The fluorescence excitation and emission wavelengths of the derivatives were lambda(ex) = 250 nm and lambda(em) = 512 nm, respectively. Linear correlation coefficients for all fatty acid derivatives are > 0.9999. Detection limits, at a signal-to-noise ratio of 3 : 1, are 24.76-98.79 fmol for the labeled fatty acids.

Pre-column derivatization of amines with 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl chloroformate followed by LC-fluorescence and LC-APCI-MS

A pre-column derivatization method for the sensitive determination of amines using the labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl chloroformate (BCIC-Cl) followed by high-performance liquid chromatography with fluorescence detection has been developed. Identification of derivatives is carried out by high performance liquid chromatography/atmospheric pressure chemical ionization (LC-APCl-MS-MS). The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent is replaced by 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl functional group, which results in a sensitive fluorescence derivatizing reagent BCIC-Cl. BCIC-Cl can easily and quickly label amines. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography and show an intense protonated molecular ion corresponding m/z [MH](+) under APCl in positive-ion mode. The collision-induced dissociation of protonated molecular ion formed a product at m/z 260 corresponding to the cleavage of CH2-OCO bond. Studies on derivatization demonstrate excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% are observed with a 3 to 4-fold molar reagent excess. In addition, the detection responses for BCIC derivatives are compared with those obtained using CEOC and FMOC as derivatization reagents. The ratios of l(BCIC)/l(CEOC) and l(BCIC)/l(FMOC) are, respectively, 1.23-3.14 and 1.25-3.08 for fluorescent (FL) responses (here, l is relative fluorescence intensity). Separation of the derivatized amines had been optimized on reversed-phase Eclipse XDB-C-8 column. Detection limits are calculated from 1.0 pmol injection, at a signal-to-noise ratio of 3, are 10.6-37.8 fmol. The mean interday accuracy ranges from 94 to 105% for fluorescence detection with the largest mean %CV < 7.5. The mean interday precision for all standards is < 6.0% of the expected concentration. Excellent linear responses are observed with coefficients of > 0.9997.

Floral closure in response to temperature and pollination in Gentiana straminea Maxim. (Gentianaceae), an alpine perennial in the Qinghai-Tibetan Plateau

Floral closure may be induced by pollination and various other factors, but is rarely studied comprehensively. Different kinds of floral closure should have various effects on reproductive fitness of plants. Two contrasting types of floral closure were observed in the flowers of Gentiana straminea Maxim. in the eastern Qinghai-Tibetan Plateau. The first type occurred prior to pollination during both gender phases, in response mainly to decreasing air temperatures. Flowers closed when decreasing temperatures approached 20 degrees C and subsequently began to reopen the following day during mid-morning when air temperatures warmed to approximately 13-15 degrees C. This kind of floral closure can protect pollen grains on either stamens or stigmas, increasing fitness of both male and female. Following pollination, permanent floral closure occurred, although there was a delay between the dates of pollination and permanent closure, during which flowers continued to show temporary closure in response to low temperature episodes. The time required for permanent, pollination-induced closure varied according to the age of the gender phase, including a prolonged time before closure if pollination occurred early in the female phase. The retaining of permanent closed flowers increased both approaching (to inflorescences) and visiting (to unpollinated flowers) frequencies of individual plants when with fewer open flowers and the persisting corolla is further beneficial for seed sets of these pollinated flowers. Thus, two separate types of floral closure, one in response to environmental cues and the other in response to the age of each gender stage, appeared to have a strong influence on reproductive fitness in this species. These results revealed a different adaptive strategy of alpine plants in the sexual reproduction assurance in addition to the well-known elevated floral longevity, dominant role of more effective pollinators and increased reproduction allocation in the arid habitats.

Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry

A pre-column derivatization method for the sensitive determination of amino acids and peptides using the tagging reagent 1,2-benzo-3,4dihydrocarbazole-9-ethyl chloroformate (BCEOC) followed by high-performance liquid chromatography with fluorescence detection has been developed. Identification of derivatives was carried out by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS/MS). The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent was replaced by 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent BCEOC. BCEOC can easily and quickly label peptides and amino acids. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. The derivatives showed an intense protonated molecular ion corresponding m/z (M + H)(+) under electrospray ionization (ESI) positive-ion mode with an exception being Tyr detected at negative mode. The collision-induced dissociation of protonated molecular ion formed a product at m/z 246.2 corresponding to the cleavage of C-O bond of BCEOC molecule. Studies on derivatization demonstrate excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% are observed with a 3-4-fold molar reagent excess. Derivatives exhibit strong fluorescence and extracted detzvatization solution with n-hexane/ethyl acetate (10:1, v/v) allows for the direct injection with no significant interference from the major fluorescent reagent degradation by-products, such as 1,2-benzo-3,4-dihydrocarbazole-9-ethanol (BDC-OH) (a major by-product), mono- 1,2-benzo-3,4-dihydrocarbazole-9-ethyl carbonate (BCEOC-OH) and bis-(1,2-benzo-3,4-dihydrocarbazole-9-ethyl) carbonate (BCEOC)(2). In addition, the detection responses for BCEOC derivatives are compared to those obtained with previously synthesized 2-(9-carbazole)-ethyl chloroformate (CEOC) in our laboratory. The ratios AC(BCEOC)/AC(CEOC) = 2.05-6.51 for fluorescence responses are observed (here, AC is relative fluorescence response). Separation of the derivatized peptides and amino acids had been optimized on Hypersil BDS C-18 column. Detection limits were calculated from 1.0 pmol injection at a signal-to-noise ratio of 3, and were 6.3 (Lys)-177.6 (His) fmol. The mean interday accuracy ranged from 92 to 106% for fluorescence detection with mean %CV < 7.5. The mean interday precision for all standards was < 10% of the expected concentration. Excellent linear responses were observed with coefficients of > 0.9999. Good compositional data could be obtained from the analysis of derivatized protein hydrolysates containing as little as 50.5 ng of sample. Therefore, the facile BCEOC derivatization coupled with mass spectrometry allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids and peptides from biological and natural environmental samples. (c) 2005 Elsevier B.V. All rights reserved.

基于CAN总线的物流拣选设备的设计

简要介绍了物流的发展现状 ,详细描述了基于CAN总线的物流拣选系统的结构、供电方式和参数设定方法 ,并为系统硬件设计中的电源转换、总线驱动和地址译码等公共电路以及总线控制器、电子标签和指示灯控制器等主要设备提供了设计方案 ,为系统软件设计中的C5 1编译、通信协议、命令类型和程序控制规划等问题给出了相应的解决方法 ,还与进口同类产品的性能和价格进行了比较

边坡工程的极限分析上限法

Theory of limit analysis include upper bound theorem and lower bound theorem. To deal with slope stability analysis by limit analysis is to approximate the real solution from upper limit and lower limit. The most used method of limit analysis is upper bound theorem, therefore it is often applied to slope engineering in many cases. Although upper bound approach of limit analysis can keep away from vague constitutive relation and complex stress analyses, it also can obtain rigorous result. Assuming the critical surface is circular slip surface, two kinematically admissible velocity fields for perpendicular slice method and radial slice method can be established according to the limit analysis of upper bound theorem. By means of virtual work rate equation and strength reduction method, the upper-bound solution of limit analysis for homogeneous soil slope can be obtained. A log-spiral rotational failure mechanism for homogeneous slope is discussed from two different conditions which represent the position of shear crack passing the toe and below the toe. In the dissertition, the author also establishes a rotational failure mechanics with combination of different logarithmic spiral arcs. Furthermore, the calculation formula of upper bound solution for inhomogeneous soil slope stability problem can be deduced based on the upper bound approach of rigid elements. Through calculating the external work rate caused by soil nail, anti-slide pile, geotechnological grid and retaining wall, the upper bound solution of safety factor of soil nail structure slope, slip resistance of anti-slide pile, critical height of reinforced soil slope and active earth pressure of retaining wall can be obtained by upper bound limit analysis method. Taking accumulated body slope as subject investigated, with study on the limit analysis method to calculate slope safety factor, the kinematically admissible velocity fields of perpendicular slice method for slope with broken slip surface is proposed. Through calculating not only the energy dissipation rate produced in the broken slip surfaces and the vertical velocity discontinuity, but also the work rate produced by self-weight and external load, the upper bound solution of slope with broken slip surface is deduced. As a case study, the slope stability of the Sanmashan landslide in the area of the Three Gorges reservoir is analyzed. Based on the theory of limit analysis, the upper bound solution for rock slope with planar failure surface is obtained. By means of virtual work-rate equation, energy dissipation caused by dislocation of thin-layer and terrane can be calculated; furthermore, the formulas of safety factor for upper bound approach of limit analysis can be deduced. In the end, a new computational model of stability analysis for anchored rock slope is presented after taking into consideration the supporting effect of rock-bolts, the action of seismic force and fissure water pressure. By using the model, not only the external woke-rate done by self-weight, seismic force, fissure water pressure and anchorage force but also the internal energy dissipation produced in the slip surface and structural planes can be totally calculated. According to the condition of virtual work rate equation in limit state, the formula of safety factor for upper bound limit analysis can be deduced.

语音意识缺陷儿童的言语知觉技能

Considerable studies find that developmental dyslexia is associated with deficits in phonological processing skills, especially phonological awareness. In order to explore the nature of phonological awareness deficits in dyslexia, researchers have begun to investigate the role of speech perception. The findings about speech perception abilities in dyslexics are inconsistent. The heterogeneity of dyslexia may be responsible for the inconsistency of findings. Considering the general suggestion that phonological awareness deficits in dyslexia are attributed to categorical perception deficits, it is more direct to examine whether children with phonological awareness difficulties or phonological dyslexia show speech categorization deficits consistently. The present study would investigate whether Chinese children with phonological awareness deficits or phonological dyslexia showed abnormal speech perception. The whole study consisted of two parts. Part I screened children with phonological-awareness deficits from Year 3 kindergartens and examined their abilities of perceiving native category continuum, nonnative category contrasts and non-speech sound series. Part II selected phonological dyslexics from an elementary school as participants, and further explored the relation between phonological deficits and speech perception. The first two experiments of Part II examined separately the abilities to label stimuli in native category continuum and brief stops in different contexts, the last experiment investigated the adaptation effects of different participant groups. The main conclusions are as follows: 1) Children with phonological dyslexia showed categorical perception deficits: they had lower consistency than controls when perceiving stimuli within phonetic categories, especially for the stimuli which were not natural sounds. 2) Children with phonological dyslexia exhibited a general difficulty of perceiving brief segments of stops from different contexts. 3) Children with phonological dyslexia did not show adaptation to repeatedly presented stimuli. Based on the present conclusions and the findings of previous studies, we suggested that the representations of sound stimuli in phonological dyslexics’ brains are different from those in normal children’s; the representations of sound stimuli in dyslexics’ cortical neural networks are more diffuse and inconsistent.

Learning Classes Correlated to a Hierarchy

Trees are a common way of organizing large amounts of information by placing items with similar characteristics near one another in the tree. We introduce a classification problem where a given tree structure gives us information on the best way to label nearby elements. We suggest there are many practical problems that fall under this domain. We propose a way to map the classification problem onto a standard Bayesian inference problem. We also give a fast, specialized inference algorithm that incrementally updates relevant probabilities. We apply this algorithm to web-classification problems and show that our algorithm empirically works well.

Synthesis of a terbium fluorescent chelate and its application to time-resolved fluoroimmunoassay

A new nonadentate ligand, N, N, N-1, N-1-[2,6-bis(3'-aminomethyl-1 1'-pyrazolyl)-4-phenylpyridine]tetrakis(acetic acid) (BPTA) for a Tb3+ fluorescent complex was synthesized. The Tb3+ complex is strongly fluorescent, having a large fluorescence quantum yield of 1.00 and very long fluorescence lifetime of 2.681 ms in 0.05 M berate buffer of pH 9.1. Streptavidin (SA) was labeled with SPTA by using its succinimidyl monoester, and the BPTA-Tb3+-labeled SA was used in sandwich-type time-resolved fluoroimmunoassay (TR-FIA) of alpha -fetoprotein (AFP) and carcinoembryonic antigen (CEA) in human sera. The Tb3+-labeled SA was also used in competitive type TR-FIA of bensulfuron- methyl (BSM) in water. The detection limits of these assays are 42 pg/mL for AFP, 70 pg/mL for CEA, and 0.4 ng/mL for BSM. In addition, a new simultaneous measurement method for AFP and CEA in a human serum sample was developed by using 4,4'-bis(1 " ,1 " ,1 " ,2 " ,2 " ,3 " ,3 " -heptafluoro-4 " ,6 " -hexanedion-6 " -yl)chlorosulfo-o-terphenyl ((BHHCT)-Eu3+-labeled anti-AFP antibody, biotinylated anti-CEA antibody, and BPTA-Tb3+-labeled SA. The concentrations of AFP and CEA in 39 human serum samples were determined, and the results were compared with those of the independently determined AFP and CEA by TR-FIA with a single-label method. A good correlation was obtained with the correlation coefficients of 0.991 for AFP and 0.994 for CEA.

What's happening in European supply-chains?

Many multinational companies are supposedly viewing Europe as one region and re-shaping their approach to the supply of products to customers. Several factors are thought to be driving this trend, notably the degree of merger and acquisitions activity; the need for improved financial performance; the pressure to reduce inventories and costs, facilitated by improvements in communication and information technology systems. All of this is in the context of European market and monetary harmonisation. This paper investigates the extent and effect of the amanegement of supply chains on a pan-European baisis by multinational business. A survey was used to examine changes, both made and anticipated. to operational strategies, processes, organisational structures and physical infrastructure across a range of businesses and industry sectors. Cost reducation, driven by the need for profit and shreholder return, was found to be the priority for developments in supply chains.Many businesses reported consolidation of manufacturing and distribution activities whilst retaining discrete country-by-country organisational structures for managing customers and markets.Logistics Service Providers were seen in a traditional role as suppliers of commodity warehousing and transport services and lacked true pan-European capability. Despite the often-vaunted concept of a pan-European business model, individual businesses wwere seen to be negotiating their own path to balancing economies of scale with customers' service needs and expectations.

An application of a multiple agent system for flexible assemble tasks

C.H. Orgill, N.W. Hardy, M.H. Lee, and K.A.I. Sharpe. An application of a multiple agent system for flexible assemble tasks. In Knowledge based envirnments for industrial applications including cooperating expert systems in control. IEE London, 1989.

Learning and Reuse of Experience in Behavior-Based Service Robots

M.H.Lee, Q. Meng and H. Holstein, ?Learning and Reuse of Experience in Behavior-Based Service Robots?, Seventh International Conference on Control, Automation, Robotics and Vision (ICARCV2002), pp1019-24, December 2002, Singapore

A dichotomic search algorithm for mining and learning in domain-specific logics

Ferr?, S. and King, R. D. (2004) A dichotomic search algorithm for mining and learning in domain-specific logics. Fundamenta Informaticae. IOS Press. To appear