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.

Determination of aliphatic amines from soil and wastewater of a paper mill by pre-column derivatization using HPLC and tandem mass spectrometry (HPLC-MS/MS)

A pre-column derivatization method for the sensitive determination of aliphatic amines using the labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) followed by HPLC with fluorescence detection and APCI/NIS identification in positive-ion mode has been developed. The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent was replaced by the 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent, BCEOC, that could easily and quickly label amines. Derivatives were stable enough to be efficiently analyzed by HPLC and showed an intense protonated molecular ion corresponding m/z [M + H](+) with APCI/MS in positive-ion mode. The collision induced dissociation of the protonated molecular ion formed characteristic fragment ions at m/z 264.1, m/z 246.0 and m/z 218.1, corresponding to the cleavages of CH2CH2O-CO, CH2CH2-OCO, and N-CH2CH2O bonds. Studies on derivatization conditions demonstrated that excellent derivatization yields close to 100% were observed with a 3 to 4-fold molar reagent excess in acetonitrile solvent, in the presence of borate buffer (pH 9.0) at 40 degrees C for 10 min. In addition, the detection responses for BCEOC derivatives were compared with those obtained with CEOC and FMOC as labeling reagents. The ratios I-BCEOC/I-CEOC and I-BCEOC/I-FMOC were, respectively, 1.40-2.76 and 1.36-2.92 for fluorescence responses (here, I was the relative fluorescence intensity). Separation of the amine derivatives had been optimized on an Eclipse XDB-C-8 column. Detection limits calculated from an 0.10 pmol injection, at a signal-to-noise ratio of 3, were 18.65-38.82 fmol (injection volume 10 mu L for fluorescence detection. The relative standard deviations for intraday determination (n = 6) of standard amine derivatives (50 pmol) were 0.0063-0.037% for retention times and 3.36-6.93% for peak areas. The mean intra-and inter-assay precision for all amines were <5.4% and 5.8%, respectively. The recoveries of amines ranged from 96 to 113%. Excellent linear responses were observed with correlation coefficients of >0.9994. The established method provided a simple and highly sensitive technique for the quantitative analysis of trace amounts of aliphatic amines from biological and natural environmental samples.

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 30 free fatty acids in two famous Tibetan medicines by HPLC with fluorescence detection and mass spectrometric identification

A simple and sensitive high-performance liquid chromatographic (HPLC) method with fluorescence detection and mass spectrometric identification has been developed for analysis of 30 long-chain and short-chain free Fatty acids (FFAs). The fatty acids were derivatized to their esters with 1-[2-(p-toluenesulfonate)ethyl]-2-phenylimidazole-[4,5-f]-9,10-phenanthrene (TSPP) in N,N-dimethylformamide (DMF) at 90 degrees C with anhydrous K2CO3 as catalyst. A mixture Of C-1-C-30 fatty acids was completely separated within 60 min by gradient elution on a reversed-phase C-8 column. Qualitative identification of the acids was performed by atmospheric-pressure chemical ionization mass spectrometry (APCI-MS) in positive-ion mode. The fluorescence excitation and emission wavelengths were 260 and 380 nm, respectively. Quantitative determination of the 30 acids in two Tibetan medicines Gentiana straminea and G. dahurica was performed. The results indicated that the medicines contained many FFAs. Linear correlation coefficients for the FFA derivatives were > 0.9991. Relative standard deviations (RSDs, n = 6) for the fatty acid derivatives were < 3%. Detection limits (at a signal-to-noise ratio of 3:1) were 3.1-38 fmol. When the fatty acid derivatives were determined in the two real samples results were satisfactory and the sensitivity and reproducibility of the method were good.

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.

HPLC-APCI-MS determination of free fatty acids in Tibet folk medicine Lomatogonium rotatum with fluorescence detection and mass spectrometric identification

A simple and sensitive method for the determination of free fatty acids (FFAs) using acridone-9-ethyl-p-toluenesulfonate (AETS) as a fluorescence derivatization reagent by high performance liquid chromatography (HPLC) has been developed. Free fatty acid derivatives were separated on an Eclipse XDB-C-8 column with a good baseline resolution and detected with the fluorescence of which excitation and emission wavelengths of derivatives were set at lambda(ex) 404 and lambda(em) 440 nm, respectively. Identification of 19 fatty acid derivatives was carried out by online post-column mass spectrometry with an atmospheric pressure chemical ionization (APCI) source under positive-ion detection mode. Nineteen FFAs from the extract of Lomatogonium rotatum are sensitively determined. The results indicate that the plant Lomatogonium rotatum is enriched with an abundance of FFAs and FFAs of higher contents, which mainly focus on even carbon atoms, C-14, C-16, and C-18. The validation of the method including linearity, repeatability, and detection limits was examined. Most linear correlation coefficients for fatty acid derivatives are > 0.9989, and detection limits (at signal-to-noise of 3: 1) are 12.3-43.7 fmol. The relative standard deviations (RSDs) of the peak areas and retention times for 19 FFAs standards are < 2.24% and 0.45%, respectively. The established method is rapid and reproducible for the separation determination of FFAs from the extract of Lomatogonium rotatum with satisfactory results.

Seasonal regulations of energetics, serum concentrations of leptin, and uncoupling protein 1 content of brown adipose tissue in root voles (Microtus oeconomus) from the Qinghai-Tibetan plateau

Survival of small mammals in winter requires proper adjustments in physiology, behavior and morphology. The present study was designed to examine the changes in serum leptin concentration and the molecular basis of thermogenesis in seasonally acclimatized root voles (Microtus oeconomus) from the Qinghai-Tibetan plateau. In January root voles had lower body mass and body fat mass coupled with higher nonshivering thermogenesis (NST) capacity. Consistently, cytochrome c oxidase activity and mitochondrial uncoupling protein-1 (UCP1) protein contents in brown adipose tissues were higher in January as compared to that in July. Circulating level of serum leptin was significantly lower in winter and higher in July. Correlation analysis showed that serum leptin levels were positively related with body mass and body fat mass while negatively correlated with UCP1 protein contents. Together, these data provided further evidence for our previous findings that root voles from the Qinghai-Tibetan plateau mainly depend on higher NST coupled with lower body mass to enhance winter survival. Further, fat deposition was significantly mobilized in cold winter and leptin was potentially involved in the regulation of body mass and thermogenesis in root voles. Serum leptin might act as a starvation signal in winter and satiety signal in summer.

Determination of five pharmacologically active compounds extracted from Rhodiola for natural product drug discovery with HPLC-APCI-MS

A rapid and sensitive liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (HPLC-APCI-MS) assay for the determination of five pharmacologically active compounds (PAC) extracted from the traditional Chinese medicine, Rhodiola , namely salidroside, tyrosol, rhodionin, gallic acid, and ethyl gallate has been developed. In this method, PAC could be baseline separated and detected with DAD at 275 nm. The validation of the method, including sensitivity, linearity, repeatability, and recovery, was examined. The linear calibration curves were acquired with correlation coefficient >0.999 and the limits of detection LOD (at a signal-to-noise ratio=3:1) were between 0.058 and 1.500 mu mol/L. It was found, that the amounts of PAC varied with different species of Rhodiola . The established method is rapid and reproducible for the separation of five natural pharmacologically active compounds from extracts of Rhodiola with satisfactory results.

双模式相关技术研究与实现

文中研究的相关跟踪技术主要应用于飞航导弹的末制导。现阶段，激光制导技术、GPS制导技术、合成孔径雷达（SAR）制导技术在我国的实际应用还不成熟，传统的电视、红外制导技术仍然具有很强的生命力，因而，基于可见光的视频相关跟踪技术具有重要的研究价值。本文主要论述的两种相关算法是：多灰度点相关（MPC）、区域模板相关（RTC）。其中多点相关（MPC）算法的跟踪灵敏度高，定位精度好，硬件实现比较方便，实时性能好；区域模板相关算法（RTC），在图像的匹配过程中，不仅考虑了目标区域的灰度特征，而且兼顾了区域里多灰度层次的位置特征、面积特征，算法具有很好的鲁棒性。文中深入研究了两种相关跟踪算法，并针对它们在实际应用中的不足，提出了有效的改善措施。最后，本文对两种相关跟踪算法进行了初步融合，一是：通过粗匹配、精匹配过程来选取目标跟踪点；二是：提出了一种度量模板更新的能量准则函数。大量的仿真实验结果表明：改进后的两种相关跟踪技术可以较好地完成一些复杂背景下的目标跟踪任务，两种算法的有效结合又进一步提高了目标跟踪的稳定性能和可靠性能。本文研究的一些相关跟踪技术已经运用到实际工程项目中。

AUV定位信号检测延时的蒙特卡洛模拟分析

在简要介绍AUV声学定位声纳接收机原理基础上,分析了CW脉冲信号在极性相关检测电路中的传输过程,建立了极性相关积分检测延时仿真分析模型。提出采用蒙特卡洛模拟方法获取检测延时的分布特征和统计参数的观点。实验结果表明蒙特卡洛模拟实验与硬件电路实验结果一致,对于解决随机性检测延时问题具有很强的能力。获得的结果可为AUV定位声纳检测门限的设定、声学测距和定位精度分析以及水声通信延时分析提供参考。

东海陆架盆地复杂天然气藏地震勘探判别技术研究

Exploration study proves that East sea shelf basin embeds abundant hydrocarbon resources. However, the exploration knowledge of this area is very low. Many problems in exploration are encountered here. One of them is that the gas reservoir of this area, with rapid lateral variation, is deeply buried. Correlation of Impendence between sandstone, gas sand and shale is very poor. Another problem is that the S/N ratio of the seismic data is very low and multiples are relatively productive which seriously affect reservoir identification. Resolution of the seismic data reflected from 2500-3000 meter is rather low, which seriously affects the application of hydrocarbon direct identification (HDI) technology. This research established a fine geological & geophysical model based on drilling、well logging、geology&seismic data of East sea Lishui area. A Q value extraction method from seismic data is proposed. With this method, Q value inversion from VSP data and seismic data is performed to determine the subsurface absorption of this area. Then wave propagation and absorption rule are in control. Field acquisition design can be directed. And at the same time, with the optimization of source system, the performance of high resolution seismic acquisition layout system is enhanced. So the firm foundation is ensured for east sea gas reservoir exploration. For solving the multiple and amplitude preserving problems during the seismic data processing, wave equation pre-stack amplitude preservation migration and wave equation feedback iteratively multiple attenuation technologies are developed. Amplitude preservation migration technology can preserve the amplitude of imaging condition and wave-field extrapolation. Multiple removing technology is independent of seismic source wavelet and velocity model, which avoiding the weakness of Delft method. Aiming at the complicated formation condition of the gas reservoir in this area, with dissecting typical hydrocarbon reservoir, a series of pertinent advanced gas reservoir seismic identification technologies such as petrophysical properties analyzing and seismic modeling technology、pre-stack/post-stack joint elastic inversion, attribute extraction technology based on seismic non-stationary signal theory and formation absorption characteristic and so on are studied and developed. Integrated analysis of pre-stack/post-stack seismic data, reservoir information, rock physics and attribute information is performed. And finally, a suit of gas reservoir identification technology is built, according to the geological and geophysical characteristics of this area. With developed innovative technologies, practical application and intergrated interpretation appraisal researches are carried out in Lishui 36-1.The validity of these technologies is tested and verified. Also the hydrocarbon charging possibility and position of those three east sea gas exploration targets are clearly pointed out.

基于匹配追踪方法的地震信号时频分析研究

Seismic signal is a typical non-stationary signal, whose frequency is continuously changing with time and is determined by the bandwidth of seismic source and the absorption characteristic of the media underground. The most interesting target of seismic signal’s processing and explaining is to know about the local frequency’s abrupt changing with the time, since this kind of abrupt changing is indicating the changing of the physical attributes of the media underground. As to the seismic signal’s instantaneous attributes taken from time-frequency domain, the key target is to search a effective, non-negative and fast algorithm time-frequency distribution, and transform the seismic signal into this time-frequency domain to get its instantaneous power spectrum density, and then use the process of weighted adding and average etc. to get the instantaneous attributes of seismic signal. Time-frequency analysis as a powerful tool to deal with time variant non-stationary signal is becoming a hot researching spot of modern signal processing, and also is an important method to make seismic signal’s attributes analysis. This kind of method provides joint distribution message about time domain and frequency domain, and it clearly plots the correlation of signal’s frequency changing with the time. The spectrum decomposition technique makes seismic signal’s resolving rate reach its theoretical level, and by the method of all frequency scanning and imaging the three dimensional seismic data in frequency domain, it improves and promotes the resolving abilities of seismic signal vs. geological abnormal objects. Matching pursuits method is an important way to realize signal’s self-adaptive decomposition. Its main thought is that any signal can be expressed by a series of time-frequency atoms’ linear composition. By decomposition the signal within an over completed library, the time-frequency atoms which stand for the signal itself are selected neatly and self-adaptively according to the signal’s characteristics. This method has excellent sparse decomposition characteristics, and is widely used in signal de-noising, signal coding and pattern recognizing processing and is also adaptive to seismic signal’s decomposition and attributes analysis. This paper takes matching pursuits method as the key research object. As introducing the principle and implementation techniques of matching pursuits method systematically, it researches deeply the pivotal problems of atom type’s selection, the atom dictionary’s discrete, and the most matching atom’s searching algorithm, and at the same time, applying this matching pursuits method into seismic signal’s processing by picking-up correlative instantaneous messages from time-frequency analysis and spectrum decomposition to the seismic signal. Based on the research of the theory and its correlative model examination of the adaptively signal decomposition with matching pursuit method, this paper proposes a fast optimal matching time-frequency atom’s searching algorithm aimed at seismic signal’s decomposition by frequency-dominated pursuit method and this makes the MP method pertinence to seismic signal’s processing. Upon the research of optimal Gabor atom’s fast searching and matching algorithm, this paper proposes global optimal searching method using Simulated Annealing Algorithm, Genetic Algorithm and composed Simulated Annealing and Genetic Algorithm, so as to provide another way to implement fast matching pursuit method. At the same time, aimed at the characteristics of seismic signal, this paper proposes a fast matching atom’s searching algorithm by means of designating the max energy points of complex seismic signal, searching for the most optimal atom in the neighbor area of these points according to its instantaneous frequency and instantaneous phase, and this promotes the calculating efficiency of seismic signal’s matching pursuit algorithm. According to these methods proposed above, this paper implements them by programmed calculation, compares them with some open algorithm and proves this paper’s conclusions. It also testifies the active results of various methods by the processing of actual signals. The problems need to be solved further and the aftertime researching targets are as follows: continuously seeking for more efficient fast matching pursuit algorithm and expanding its application range, and also study the actual usage of matching pursuit method.

尾波干涉法精确测量地震波速变化的方法及实验研究

The coda of seismic waves consists of that part of the signal after the directly arrivials. In a finite medium, or in one that is strongly heterogeneous, the coda is dominated by waves which have repeatedly sampled the medium. Small changes in a medium which may have no detectable influence on the first arrivals are amplified by this repeated sampling and may thus be detectable in the coda. Because of this, coda wave is widely used in detecting micro variations in medium。 In this paper, we give a general view of the theory and application of coda wave, especially coda wave interferometry. We focus on discussing the application of coda wave interferometry on data source of active situ experiment。 First, we apply coda wave interferometry in a short time period situ experiment which last for three days. We also apply the method of coda wave interferometry in a situ experiment which last for one month. Daily circle variations of seismic velocity around the experiment site were obtained, and we also observed that the velocity variations in the experiment site have a significant correlation with the environment factors, including air temperature, barometric pressure, solid earth tide and the level of rainfall. We find that the velocity variation during this period is up to 10-3. The relationship between velocity variation and changes in air temperature, barometric pressure and solid earth tide was analyzed with least square linear fitting .The velocity has no dependence on the air temperature. But velocity has a change of 10-6--10-7 when the barometer or earth tide change per Pa. Generally, we conclude the work and results of previous researchers, and we also display our works and results. We hopes to contribute to the future research of coda wave interferometry.

基于分形和统计理论的地层非均质研究

Basin-scale heterogeneity contains information about the traces of the past sedimentary cycle and tectonic process, and has been a major concern to geophysicists because of its importance in resource exploration and development. In this paper, the sonic data of 30 wells of Sulige field are used to inverse the power-law spectra slope and correlation length which are measures of the heterogeneity of the velocity of the log using fractal and statistic correlation methods. By taking the heterogeneity parameters of different wells interpolated, we get power law spectra slope and correlation length contours reflecting the stratum heterogeneity. Then using correlation and gradient, we inverse the transverse heterogeneity of Sulige field. Reservior-scale heterogeneity influnce the distribution of remaining oil and hydrocarbon accumulation. Using wavelet modulus maximum method to divide the sedimentary cycle using Gr data, therefore we can calculate the heterogeneity parameter in each layer of each log. Then we get the heterogeneity distribution of each layer of Sulige field. Finally, we analyze the relation between the signal sigularity and the strata heterogeneity, and get two different sigularity profiles in different areas.

河流-三角洲储层自动对比微相自动识别原理方法和软件系统

Because of the complexity and particularity, especially the result is more depend on the expert' s experience, the calculate method which is based on the simplicity mathematical model can hardly have any effective role in the oilfield .The coalescent method of artificial intelligence and signal manage in the correlation of reservoir use log curve has been put forward.in this paper. Following the principle of "controlled by classification and correlation by deposit gyration ". The system of correlation has been setup, which can identify "standard layer" first by the improved method of gray connection system, and then on the basis of identified "standard layer", interpret the fault, and last identify the layer in the reservoir. A effective method of "the consistent character of a reservoir "has been adopt to solved the puzzle of interpret the fault. On the basis of sedimentary theory and the quantity analysis of log curve shape of different type microfacies, a serial of different type micofacies' s models has been build that use eight optimized parameters, five of eight rationed parameters being used to describe microfacies with log curve, the distribution area of every parameters for the microfacies has been give. Because the classical math can only be used in the areas that principles are very clearly, not be fit for the description of geology character, so The fuzzy math integrate judgment has been adopt in the using log curve to determine microfacies; the accordance ration is 85 percent. A set of software has been programmed which is on the system of Windows. the software has the integration function of data process, auto-contrast reservoir layer, determination of microfacies using log curve, character the connectivity of sandstones and plotting of geology map. Through the application, this system has high precision and has become a useful tool in the study of geology.