棘胸蛙不同发育阶段肌肉差异蛋白的质谱分析

利用串联质谱对棘胸蛙的不同发育阶段个显著差异的肌肉蛋白质进行分析,根据质谱数据搜索蛋白数据库成功获得蛋白信息.构建了适应于质谱鉴定的棘胸蛙肌肉蛋白样处理方法,为进一步深入研究影响棘胸蛙发育的蛋白质奠定了基础.     

微量含硒蛋白检测技术进展

生物必需微量元素硒在体内主要通过硒蛋白发挥作用,因此微量含硒蛋白的高灵敏检测和识别是验证生物信息学所预测的硒蛋白和开展硒蛋白组学研究的重要手段,也是硒蛋白功能研究的先决条件。文章重点介绍了近年迅速发展的激光剥蚀电感耦合等离子体质谱技术原理及其对凝胶电泳分离后蛋白质硒的检测,简要综述了凝胶电泳分离后其他质谱技术在含硒蛋白分析鉴定中的应用,包括毛细管高效液相色谱/纳升高效液相色谱-电感耦合等离子体质谱对含硒多肽的分离检测、基质辅助激光解吸离子化-飞行时间-质谱对含硒蛋白的识别、以及纳升高效液相色谱-电喷雾-串联质谱对含硒蛋白的鉴定等内容,指出了微量含硒蛋白检测中存在的问题和发展方向。     

新疆木垒香阿魏成分提取及分析

超临界CO2流体萃取、溶剂二氯甲烷和水蒸气蒸馏等不同方法对新疆木垒香阿魏根进行提取,气相色谱-质谱联用分析比较其中成分;从该种植物中共得到42个峰,其中鉴定出28个化合物;三种方法中超临界CO2流体萃取方法优于传统方法,三者中主要挥发性成分愈创木醇的含量都很高。     

SPME/GC/MS分析小一点红挥发性化学成分

利用固相微萃取/气相色谱/质谱(SPME/GC/MS)联用技术对小一点红挥发性成分进行研究。共鉴定出25种化学成分,占挥发性总成分的91.99%。小一点红主要挥发性化学成分是β-月桂烯(51.18%),3,7,11,15-四甲基-2-十六烯-1-醇(21.55%),β-水芹烯(8.42%),n-十六酸(2.48%)等。     

SPME/GC/MS分析荷花玉兰挥发性成分

利用固相微萃取/气相色谱/质谱(SPME/GC/MS)联用技术对荷花玉兰挥发性成分进行研究。共鉴定出48个化学成分,占挥发性总成分的85.76%。荷花玉兰主要挥发性化学成分是反式-香叶醇(38.11%),苯甲醇(4.99%),3,7-二甲基-1,6-辛二烯-3-醇(4.33%),松香芹酮(3.24%)等。     

新型白杨素-7-氨基磷酸酯衍生物的合成与波谱学研究

以5, 7-二羟基黄酮和磷酰化氨基酸酯为原料,利用磷酰化反应将磷酰化氨基酸酯引入到白杨素中,得到了一种新型的白杨素-7-氨基磷酸酯衍生物. 通过DEPT、¹H-¹H COSY、¹³C-¹H HSQC和¹³C-¹H HMBC等2D NMR核磁共振技术对该化合物的¹H、¹3C NMR谱的信号进行了全归属和较详细的解析,并探讨了其ESI MS/MS质谱裂解规律.      

四氯蔗糖及其衍生物的NMR碳氢化学位移完全解析及结构研究

4,6,1′,6′-四氯蔗糖及其两个单酯化衍生物4,6,1′,6′-四氯蔗糖-3-乙酯和4,6,1′,6′-四氯蔗糖-2-乙酯是在合成三氯蔗糖的过程中得到的3种副产物，应用质谱（MS）和核磁共振（NMR）中的¹H-¹H COSY、HSQC等多种二维谱技术对它们的结构进行了研究，报道了四氯蔗糖及其两个单酯衍生物的合成及NMR碳氢化学位移完全解析，并对其氯化产物的立体化学进行了讨论.     

黄芩素的分离纯化与结构表征

以聚酰胺为柱层析吸附剂,以乙醇-氯仿混合溶剂梯度洗脱,从中药黄芩的甲醇提取液中分离出有效成分黄芩素的粗品。 通过对有效成分进行重结晶等纯化处理,得到了一种黄色短棱柱晶状产物。 利用红外光谱(FTIR)、紫外-可见光谱(UV-Vis)、质谱(MS)、氢谱(1H-NMR)及碳谱(13C-NMR)等分析测试手段,对产物的组成和结构进行了表征。 红外光谱测定结果表明,产物的结构中含有共轭芳香体系和单取代苯;紫外-可见光谱和质谱表明,产物为黄酮而非黄酮苷化合物,A环上含有三个酚羟基, B环无羟基和其他基团取代;核磁共振氢谱及碳谱表明,产物的结构中含15个碳原子, 三组羟基质子。 同时,文章还对所有1H-NMR和13C-NMR信号进行了归属,对FTIR光谱特征吸收峰所对应的官能团及其振动形式进行了指认,给出了化合物及碎片离子在质谱中的裂解方式,最终将黄色短棱柱晶状产物鉴定为5,6,7-三羟基黄酮,即黄芩素,分子式为C₁₅H₁₀O₅。     

根区增温对玉米幼苗主要代谢物傅里叶红外光谱特性及叶绿素含量的影响

为深入研究根区温度对玉米幼苗主要代谢物以及叶绿素含量的影响,试验设置常温(24℃)、中温(30℃)和高温(36℃)三个根区温度水平,以SD609(保绿型)和SD902(衰老型)两个不同抗逆型玉米品种为样品,采用傅里叶变换红外光谱仪(TensorⅡ,德国布鲁克)测定玉米幼苗根、茎、叶三个器官的光谱特征。结果表明:不同根区温度处理对玉米幼苗各个器官有机物含量有显著影响,但影响程度不同,大体表现为根茎叶;玉米幼苗根、茎、叶在3 330, 2 927, 1 639, 1 515, 1 350, 1 250和1 055 cm~(-1)等波数处透射率均受根区温度的影响,且在根区中温条件下各器官上述波数处透射率较低,根区高温条件下透射率较高,在1 055 cm~(-1)处各处理差异最为明显;不同抗逆型品种在根区增温条件下FTIR光谱特征有较大差异,抗逆性较差的衰老型品种(SD902)在不同根区温度处理下FTIR光谱特征差异较大,叶绿素含量受根区增温影响较大,而抗逆性较好的保绿型品种(SD609)不同处理间差异较小,叶绿素含量受根区增温影响较小;根区温度对不同玉米品种根系活力及叶绿素含量影响均表现为中温(30℃)对照(24℃)高温(36℃)。因此,适当提高根区温度可以显著提高玉米幼苗根系活力、叶绿素、碳水化合物尤其是多糖含量,以及茎秆和叶片中蛋白质、核酸含量,但温度过高会使多糖、脂类、叶绿素含量以及根系活力迅速下降;与SD609相比, SD902对根区温度反应较为敏感,在根区高温条件下,玉米内源物质合成受阻,而SD609能保持相对较高的多糖及叶绿素含量,对根区高温条件有相对较好的适应性。根区温度对玉米幼苗根系作用最大,可通过影响根系吸收、物质合成及物质转运功能,进而影响茎秆、叶片有机物以及叶绿素含量。     

苦参生物成碱的二维核磁共振谱(2D-NMR)研究Ⅰ.中国东北产苦参中的N-甲基金雀儿碱与氧化槐根碱的2D-NMR研究

由我国东北产的苦参中分离出七种生物碱。由MS、IR及2D-NMR鉴定为:(+)-苦参碱、(+)-氧化苦参碱、5α-羟基苦参碱、N-甲基金雀儿碱、9α-羟基苦参碱及氧化槐根碱。另一个生物碱(6)的化学结构尚在鉴定之中。本课题采用了二种2D-NMR技术,H-H相关谱(300、400及500MHz)及C-H相关谱(H:300MHz;C:75MHz)。先发表其中两个生物碱的2D-NMR,其它将陆续发表。     

The limits of specificity: An experimental analysis with RNA aptamers to MS2 coat protein variants

It has been hypothesized that selections for aptamers with high affinity for a given target molecule will of necessity identify aptamers that have high specificity for that target. We have attempted to assess this hypothesis by selecting aptamers that can bind to MS2 coat protein or to single- or double-substitution variants of the coat protein. Some aptamers selected to bind MS2 coat protein or its variants were mildly specific for their cognate targets, discriminating by two- to fourfold against closely related proteins. Specificity determinants on both the coat proteins and the aptamers could be identified. However, many aptamers could readily bind to each of the different coat proteins. The identification of such aptamer 'generalists belies the proposed relationship between the affinities and specificities of selected RNA ligands. These results imply that, while aptamers may not finely discriminate between closely related targets, neither will their binding be negated by mutations in targets. Aptamer pharmaceuticals may therefore better resist the evolution of resistance.     

Full utilization of a mass spectrometer using on-demand sharing with multiple LC units

The applicability of liquid chromatography-mass spectrometry (LC/MS) is often limited by throughput. The sharing of a mass spectrometer with multiple LCs significantly improves throughput; however, the reported systems have not been designed to fully utilize the MS duty cycle, and as a result to achieve maximum throughput. To fully utilize the mass spectrometer, the number of LC units that a MS will need to recruit is application dependent and could be significantly larger than the current commercial or published implementations. For the example of a single analyte, the number may approach the peak capacity to a first degree approximation. Here, the construction of a MS system that flexibly recruits any number of LC units demanded by the application is discussed, followed by the method to port a previously developed LC/MS method to the system to fully utilize a mass spectrometer. To demonstrate the performance and operation, a prototypical MS system of eight LC units was constructed. When 1-min chromatographic separations were performed in parallel on the eight LCs of the system, the average LC/MS analysis time per sample was 10.5 s when applied to the analysis of samples in 384-well plate format. This system has been successfully used to conduct large-volume biochemical assays with the analysis of a variety of molecular entities in support of drug discovery efforts. Allowing the recruitment of the number of LC units appropriate for a given application, this system has the potential to be a plug-and-play system to fully utilize a mass spectrometer. Copyright ? 2012 John Wiley & Sons, Ltd.     

Biophysical characteristics of cells cultured on cholesteryl ester liquid crystals

This study aimed at examining the biophysical characteristics of human derived keratinocytes (HaCaT) cultured on cholesteryl ester liquid crystals (CELC). CELC was previously shown to improve sensitivity in sensing cell contractions. Characteristics of the cell integrin expressions and presence of extracellular matrix (ECM) proteins on the liquid crystals were interrogated using various immunocytochemical techniques. The investigation was followed by characterization of the chemical properties of the liquid crystals (LC) after immersion in cell culture media using Fourier transform infrared spectroscopy (FTIR). The surface morphology of cells adhered to the LC was studied using atomic force microscopy (AFM). Consistent with the expressions of the integrins α2, α3 and β1, extracellular matrix proteins (laminin, collagen type IV and fibronectin) were found secreted by the HaCaT onto CELC and these proteins were also secreted by cells cultured on the glass substrates. FTIR analysis of the LC revealed the existence of spectrum assigned to cholesterol and ester moieties that are essential compounds for the metabolizing activities of keratinocytes. The immunostainings indicated that cell adhesion on the LC is mediated by self-secreted ECM proteins. As revealed by the AFM imaging, the constraint in cell membrane spread on the LC leads to the increase in cell surface roughness and thickness of cell membrane. The biophysical expressions of cells on biocompatible CELC suggested that CELC could be a new class of biological relevant material.     

Determination of levamisole in feeds by liquid chromatography coupled to electrospray mass spectrometry on an ion trap

Test methods have to be developed by laboratories for official control to monitor possible misuse of veterinary drugs in animal productions, also through feeding stuff. A novel method for identification and quantification of levamisole in feeds by liquid chromatography coupled to electrospray mass spectrometry in an ion trap (LC/ESI-MS/MS) is herein described; after a single-step cleanup by liquid-liquid extraction from the feed and separation by reversed-phase liquid chromatography, levamisole was determined and unambiguously confirmed by tandem mass spectrometry, on the basis of two product ions. The method was in-house validated, according to the Regulation 882/2004/EC, evaluating trueness, repeatability, within-laboratory reproducibility, ruggedness, specificity, and the limit of quantification (LOQ). The method is reliable and specific for complete and complementary feeds for pigs, cattle, rabbits and poultry; very good mean recoveries (higher than 92 %) and precision (RSD values?相似文献   

Identification and structural characterization of in vivo metabolites of ketorolac using liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS)

In vivo metabolites of ketorolac (KTC) have been identified and characterized by using liquid chromatography positive ion electrospray ionization high resolution tandem mass spectrometry (LC/ESI-HR-MS/MS) in combination with online hydrogen/deuterium exchange (HDX) experiments. To identify in vivo metabolites, blood urine and feces samples were collected after oral administration of KTC to Sprague-Dawley rats. The samples were prepared using an optimized sample preparation approach involving protein precipitation and freeze liquid separation followed by solid-phase extraction and then subjected to LC/HR-MS/MS analysis. A total of 12 metabolites have been identified in urine samples including hydroxy and glucuronide metabolites, which are also observed in plasma samples. In feces, only O-sulfate metabolite and unchanged KTC are observed. The structures of metabolites were elucidated using LC-MS/MS and MS(n) experiments combined with accurate mass measurements. Online HDX experiments have been used to support the structural characterization of drug metabolites. The main phase I metabolites of KTC are hydroxylated and decarbonylated metabolites, which undergo subsequent phase II glucuronidation pathways.     

The detection of nicotine in a Late Mayan period flask by gas chromatography and liquid chromatography mass spectrometry methods

Several ancient Mayan vessels from the Kislak Collection of the US Library of Congress were examined for the presence of alkaloids. One of them, a codex-style flask, bears a text that appears to read yo-'OTOT-ti 'u-MAY, spelling y-otoot 'u-may 'the home of its/his/her tobacco'. Samples extracted from this Late Classic period (600 to 900?AD) container were analyzed by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS) methods. Nicotine was identified as the major component of the extracts. LC/MS analyses also yielded signals due to nicotine mono-oxides. The identities of the compounds were determined by comparison of the chromatographic and/or mass spectral characteristics with those from standards and literature data. High-resolution high mass accuracy tandem mass spectrometry (MS/MS) spectra of protonated nicotine and nicotine mono-oxides were measured to verify and to correct previous product ion assignments. These analyses provided positive evidence for nicotine from a Mayan vessel, indicating it as a likely holder of tobacco leafs. The result of this investigation is the first physical evidence of tobacco from a Mayan container, and only the second example where the vessel content recorded in a Mayan hieroglyphic text has been confirmed directly by chromatography/mass spectrometry trace analysis.     

深度神经网络和高光谱显微图像的二维材料纳米片识别

近年来,二维材料由于其独特的性质而受到了广泛关注。在制备二维层状晶体的各种方法中,机械剥离法获得的薄层二维材料晶体质量高,适用于基础研究及性能演示。然而用机械剥离法从衬底上获得的材料具有一定的随机性,可能包含了少许相对较厚的部分。实现对这些二维薄层材料有效、快速且智能化的表征有利于促进二维材料性能的进一步研究。提出了一种基于深度学习的表征方法,通过搭建的编解码结构的卷积神经网络语义分割算法,可以根据光学显微镜图像进行分割和快速识别二维材料纳米片。卷积神经网络作为深度学习在图像处理领域中的典型算法,能够对光学显微镜图像中的复杂信息进行特征提取。首先采用机械剥离制备MoS₂纳米片样本,通过光学显微镜采集高光谱图像并对样本进行标记,根据样本的厚度范围标记出不同的区域,对标记后的图像进一步处理,包括图像的颜色校准和剪切操作,得到用于网络训练和测试的数据集。针对光学图像中二维纳米薄片存在的低对比度、碎裂等特点,编码时加入残差结构和金字塔池化模型,有助于特征信息的提取;解码时融合编码路径中提取的浅层特征信息,以提高网络分割精度。实验中采用带权重的交叉熵损失函数解决类别数量不平衡问题和采用数据增强扩大数据集。对训练后的网络测试结果表明,模型像素精度为97.38%,平均像素精度为90.38%,均交并比为75.86%。之后通过迁移学习成功地对剥离的单层和双层石墨烯纳米片样本进行了识别,均交并比达到了81.63%,表明该方法具有普适性。通过MoS₂和石墨烯纳米片的识别演示,实现了深度学习在二维材料的光学显微镜图像中的成功应用。该方法有望在更多的二维材料上得到扩展并突破自动动态处理光学显微镜图像的问题,同时为其他纳米材料的高光谱图像处理提供参考。     

基于全血可见吸收光谱的物种鉴别

血液及血液制品是一种特殊的商品,在流通过程中需要严加管控。传统的方法通常为应用特征抗体检测人血红蛋白,或应用液相色谱(HPLC)、质谱(MS)等方法对血液物种进行鉴别,鉴别过程中外界环境和血液样本会发生接触。为了防止血液样本和外界环境相互污染,因此需要发展非接触式的血液物种鉴别技术。设计了一种基于吸收光谱的血液物种鉴别方法。实验测试了人与牛全血样本共24份,每种样本各10份,另有4份标记为未知样本。装有血液样本的PET管置于积分球内,使用超连续谱光源对积分球进行照射,测量420～740 nm范围内匀光后的吸收光谱,每个样品测试5次取平均。实验测试了使用主成分分析法对测得的原始光谱数据进行降维。并对降维后的数据应用二维正态分布进行拟合。通过样本在置信空间内的概率分布或到已知分类中心的距离对未知样本的物种进行鉴别。可以得出结论,应用PCA和二维正态分布拟合的方法,能够较为精确地实现对全血的可见吸收光谱物种鉴别。