Identification of triacylglycerol using automated annotation of high resolution multistage mass spectral trees

High complexity of identification for non-target triacylglycerols (TAGs) is a major challenge in lipidomics analysis. To identify non-target TAGs, a powerful tool named accurate MSⁿ spectrometry generating so-called ion trees is used. In this paper, we presented a technique for efficient structural elucidation of TAGs on MSⁿ spectral trees produced by LTQ Orbitrap MSⁿ, which was implemented as an open source software package, or TIT. The TIT software was used to support automatic annotation of non-target TAGs on MSⁿ ion trees from a self-built fragment ion database. This database includes 19108 simulate TAG molecules from a random combination of fatty acids and corresponding 500582 self-built multistage fragment ions (MS ≤ 3). Our software can identify TAGs using a “stage-by-stage elimination” strategy. By utilizing the MS¹ accurate mass and referenced RKMD, the TIT software can discriminate unique elemental composition candidates. The regiospecific isomers of fatty acyl chains will be distinguished using MS² and MS³ fragment spectra. We applied the algorithm to the selection of 45 TAG standards and demonstrated that the molecular ions could be 100% correctly assigned. Therefore, the TIT software could be applied to TAG identification in complex biological samples such as mouse plasma extracts.     

微藻脂肪合成与代谢调控

随着能源与环境问题的日益严峻,利用微藻生产生物柴油已经成为研究者们关注的焦点。与传统油料作物相比,微藻具有生长速度快、含油量高、不占用耕地等优势,是极具潜力的生物燃料生产原料。虽然许多微藻在压力条件下会在细胞内积累脂肪,特别是中性脂肪三酰甘油(TAG),它是生产生物柴油的主要原料,但目前对于微藻脂肪的合成和代谢调控还了解的很少。为了更好地理解和操纵微藻脂肪代谢以增强脂肪积累,本文综述了微藻脂肪合成与代谢调控的研究进展,包括TAG生物合成途径,提高脂肪积累的生化调控和基因工程策略,阐述了营养控制对脂肪积累的影响,总结了增强脂肪酸合成途径、增强Kennedy途径、调控TAG旁路途经、抑制脂肪合成的竞争途径、抑制脂肪的分解代谢等5种基因工程策略,同时也对微藻脂肪代谢研究的发展进行了展望。     

Tb3Al5O12∶Ce3+发光粉的微波辅助合成和性能表征

采用微波辅助合成-高温热处理制备了铈掺杂铽铝石榴石(Tb3Al5O12∶Ce3 ,简称TAG∶Ce)发光粉。用XRD、SEM、激发和发射光谱等对粉末的晶型、形貌以及发光性能进行了表征。结果表明,发光粉晶粒清晰、表面光滑,粒径在2μm以下。TAG∶Ce的激发光谱主激发峰在467nm,330nm左右存在次激发峰,375nm附近存在两个强度较弱的峰;发射光谱峰值波长为550nm,与传统固相法制备的TAG∶Ce基本一致。与传统高温固相反应法相比,此方法具有高效、节能、环保等优点。通过分析反应历程,证明了Tb2O3-Al2O3系统中Al离子的分扩散系数大于Tb离子的。     

Chiral recognition by enantioselective liquid chromatography: Mechanisms and modern chiral stationary phases

An overview of the state-of-the-art in LC enantiomer separation is presented. This tutorial review is mainly focused on mechanisms of chiral recognition and enantiomer distinction of popular chiral selectors and corresponding chiral stationary phases including discussions of thermodynamics, additivity principle of binding increments, site-selective thermodynamics, extrathermodynamic approaches, methods employed for the investigation of dominating intermolecular interactions and complex structures such as spectroscopic methods (IR, NMR), X-ray diffraction and computational methods. Modern chiral stationary phases are discussed with particular focus on those that are commercially available and broadly used. It is attempted to provide the reader with vivid images of molecular recognition mechanisms of selected chiral selector–selectand pairs on basis of solid-state X-ray crystal structures and simulated computer models, respectively. Such snapshot images illustrated in this communication unfortunately cannot account for the molecular dynamics of the real world, but are supposed to be helpful for the understanding. The exploding number of papers about applications of various chiral stationary phases in numerous fields of enantiomer separations is not covered systematically.     

Enantio- and chemo-selective HPLC separations by chiral-achiral tandem-columns approach: the combination of CHIROBIOTIC TAG and SCX columns for the analysis of propionyl carnitine and related impurities

We describe a new tandem-columns chiral-achiral HPLC arrangement by using a chiral column (CHIROBIOTIC TAG) connected in series with an achiral column (Spherisorb S5 SCX), based on a strong cationic exchange mechanism; this approach is very useful for the analysis of chiral molecules, containing cationic groups in their structures. We used this special combination to develop an easy and convenient procedure for the enantio- and chemo-selective dosage of propionyl L-carnitine (1) and relative impurities (2-6), which allowed for the simultaneous separation and quantitation within 30 min. Under the best chromatographic conditions (acetonitrile-10 mM sodium dihydrogen phosphate 65:35, v/v (pHa 6.80) as the mobile phase and UV detection at 205 nm], all the individual peaks were well separated. The applicability of the method, fully validated, was demonstrated by the analysis of a pharmaceutical batch of propionyl L-carnitine, where we found the following contents: 98.5% for 1 (drug substance); 0.15% for 3; 0.1% for 5 and 0.2% for 6. The enantiomeric excess (e.e.%) measured for the drug substance was 98.9%. Finally, a single mixed-bed column, packed with a binary mixture of the chiral and achiral phases, in a 1:1 ratio, gave similar chromatographic results as the tandem-columns approach, and thus, offered an easy alternative solution to the separation of the considered mixture.     

Deep-tissue confocal imaging of the central projections of ovipositor sensory afferents in the Egyptian cotton leafworm,Spodoptera littoralis

The pre-ovipositon behavior of moths is largely dependent upon the cues that a gravid female perceives while assessing potential oviposition sites. Assessment of such sites is accomplished, at least in part, by mechanosensory and gustatory sensilla located on the ovipositor whose sensory neurons project into the terminal abdominal ganglion (TAG). Using anterograde backfill staining, confocal laser scanning microscopy, and three dimensional reconstruction, we traced and analyzed the central projections of the sensory neurons housed in the sensilla located on the ovipositor papillae and explored the neuropilar composition of the TAG in the Egyptian cotton leafworm, Spodoptera littoralis. The TAG consists of three fused neuromeres (6–8th Ner) associated with the 6–8th abdominal segments. Within the TAG, and specifically in the 8th neuromere, four unstructured neuropilar compartments are present; the dorso-ipsilateral motor neuropil (MN), the medio-ipsilateral mechanosensory neuropil (MchN), the medio-ipsilateral small gustatory neuropil (GN), and the medio-contralateral posterior ovipositor glomerulus (Og). The Og appears quite compact, with a hollow core free of terminal arborizations. The MchN is further subdivided into 4 unstructured glomeruli in the 8th neuromere, whose afferents are subsequently extended into 3 glomeruli in the 7th and 6th neuromeres. Few neurites of the Og are populated with large dense varicosities reminiscent of neurosecretory vesicles. Given that all ovipositor nerves converge into a common ganglionic center, the TAG, we assume that this ganglion may be a center for coordination of oviposition behaviors, including movements of the ovipositor during assessment of oviposition substrates and egg laying in S. littoralis.     

Olive oil quantification of edible vegetable oil blends using triacylglycerols chromatographic fingerprints and chemometric tools

The present work studies the effectiveness of the use of triacylglycerols (TAGs) for the quantification of olive oil in blends with vegetable oils. The determinations were obtained using high-performance liquid chromatography (HPLC) coupled to a Charged Aerosol Detector (CAD), in combination with Partial Least Squares (PLS) regression and using interval PLS (iPLS) for variable selection.Results revealed that PLS models can predict olive oil concentrations with reasonable errors. Variable selection through iPLS did not improve predictions significantly, but revealed the chemical information important in the chromatogram to quantify olive oil in vegetable oil blends.     

反相高效液相色谱法同时测定发酵乳中的蛋白糖和氨基酸

介绍了同时测定发酵乳中蛋白糖和氨基酸的ＨＰＬＣ方法。样品经甲醇除去蛋白,异硫氰酸苯衍生,ＲＰ－ＨＰＬＣ法测定。蛋白糖回收率为９４．２％～９８．７％,精密度以相对标准偏差表示为２．１５％～３．７４％。     

Supercritical fluid chromatography as a tool for enantioselective separation; A review

Supercritical fluid chromatography (SFC) has become popular in the field of enantioselective separations. Many works have been reported during the last years. This review covers the period from 2000 till August 2013. The article is divided into three main chapters. The first one comprises a basic introduction to SFC. The authors provide a brief explanation of general principles and possibilities of this method. The advantages and drawbacks are also listed. Next part deals with chiral separation systems available in SFC, namely with the commonly used chiral stationary phases. Properties and interaction possibilities of the chiral separation systems are described. Recent theoretical papers are emphasized in this chapter. The last part of the paper gives an overview of applications of enantioselective SFC in analytical chemistry, in both analytical and preparative scales. Separation systems and conditions are summed up in tables so that they provide a helpful tool for analysts who search for a particular method of analysis.     

Lipid analysis by thin-layer chromatography--a review of the current state

High-performance thin-layer chromatography (HPTLC) is a widely used, fast and relatively inexpensive method of separating complex mixtures. It is particularly useful for smaller, apolar compounds and offers some advantages over HPLC. This review gives an overview about the special features as well as the problems that have to be considered upon the HPTLC analysis of lipids. The term "lipids" is used here in a broad sense and comprises fatty acids and their derivatives as well as substances related biosynthetically or functionally to these compounds. After a short introduction regarding the stationary phases and the methods how lipids can be visualized on an HPTLC plate, the individual lipid classes will be discussed and the most suitable solvent systems for their separation indicated. The focus will be on lipids that are most abundant in biological systems, i.e. cholesterol and its derivates, glycerides, sphingo- and glycolipids as well as phospholipids. Finally, a nowadays very important topic, the combination between HPTLC and mass spectrometric (MS) detection methods will be discussed. It will be shown that this is a very powerful method to investigate the identities of the HPTLC spots in more detail than by the use of common staining methods. Future aspects of HPTLC in the lipid field will be also discussed.