Precise and global identification of phospholipid molecular species by an Orbitrap mass spectrometer and automated search engine Lipid Search

In the present research, we have established a new lipidomics approach for the comprehensive and precise identification of molecular species in a crude lipid mixture using a LTQ Orbitrap mass spectrometer (MS) and reverse-phase liquid chromatography (RPLC) combination with our newly developed lipid search engine “Lipid Search”. LTQ Orbitrap provides high mass accuracy MS spectra by Fourier-transform (FT) mass spectrometer mode and can perform rapid MSⁿ by ion trap (IT) mass spectrometer mode. In this study, the negative ion mode was selected to detect fragment ions from phospholipids, such as fatty acid anions, by MS2 or MS3. We selected the specific detection approach by neutral loss survey-dependent MS3, for the identification of molecular species of phosphatidylcholine, sphingomyelin and phosphatidylserine. Identification of molecular species was performed by using both the high mass accuracy of the mass spectrometric data obtained from FT mode and structural data obtained from fragments in IT mode. Some alkylacyl and alkenylacyl species have the same m/z value as molecular-related ions and fragment ions, thus, direct acid hydrolysis analysis was performed to identify alkylacyl and alkenylacyl species, and then the RPLC–LTQ Orbitrap method was applied. As a result, 290 species from mouse liver and 248 species from mouse brain were identified within six different classes of phospholipid, only those in manually detected and confirmed. Most of all manually detected mass peaks were also automatically detected by “Lipid Search”. Adding to differences in molecular species in different classes of phospholipids, many characteristic differences in molecular species were detected in mouse liver and brain. More variable number of saturated and monounsaturated fatty acid-containing molecular species were detected in mouse brain than liver.     

A level set projection model of lipid vesicles in general flows

A new numerical method to model the dynamic behavior of lipid vesicles under general flows is presented. A gradient-augmented level set method is used to model the membrane motion. To enforce the volume- and surface-incompressibility constraints a four-step projection method is developed to integrate the full Navier–Stokes equations. This scheme is implemented on an adaptive non-graded Cartesian grid. Convergence results are presented, along with sample two-dimensional results of vesicles under various flow conditions.     

Progress in liquid crystalline dispersions: Cubosomes

Dispersed particles of bicontinuous cubic liquid crystalline phase, cubosomes, are self-assembled nanostructured particles that can be formed in aqueous lipid and surfactant systems. Contributions to cubosome research have come from the fields of biology, material science, medicine, and mathematics and much is known about their formation and properties. At the center of much of the discovery and innovation is the technique of cryo-transmission electron microscopy. Most of the research into cubosomes is motivated by potential applications in drug delivery and material synthesis although no commercialized product based on cubosomes is known. Recent advances in understanding and use of cubosomes are discussed in the context of some of the more promising application areas and the opportunities for microscopy techniques to make unique contributions to these areas.     

Characterization of lipid components of Melanorrhoea usitata lacquer sap

The lipid component of Melanorrhoea usitata lacquer sap isolated by acetone was analyzed and compared to synthesized ω-phenylalkylcatechols and ω-phenylalkylphenols. In addition, laccol and urushiol analogues synthesized in our laboratory were used as standard materials to analyze the lipid component of the Myanmar lacquer sap. The GC and GC/MS measurements confirmed the results of Kumanotani and Du that neither ω-phenylalkylcatechol nor ω-phenylalkylphenol exist in the lacquer saps from Rhus vernicifera and R. succedanea.     

甲酸处理对微藻油脂提取的影响

采用易于回收的有机酸处理微藻细胞,以微藻Chlorella protothecoides为原料,系统研究了甲酸处理对微藻油脂提取的影响.结果表明,甲酸能有效破碎微藻细胞,最终油脂得率和脂肪酸甲酯(FAME)得率均与传统无机酸热法相当.甲酸浓度、温度和处理时间对油脂提取有显著影响.当甲酸浓度为88%,液固比为6 m L/g,100℃下处理30 min,Chlorella protothecoides油脂得率为49.2%,FAME得率高达92.1%.甲酸处理提取到的微藻油脂脂肪酸组成与无机酸热法结果无明显区别.Chlorella protothecoides油脂主要含有5种脂肪酸,其中C18∶1含量最高.实验还考察了甲酸处理对Nannochlorum sp和Nannochloropsis oceanic的适用性,结果表明,对于不同微藻原料,甲酸处理提取到的微藻油脂得率和FAME得率均与无机酸热法结果相当.     

Standard operation protocol for analysis of lipid hydroperoxides in human serum using a fully automated method based on solid-phase extraction and liquid chromatography-mass spectrometry in selected reaction monitoring

Standard operating procedures (SOPs) are of paramount importance in the analytical field to ensure the reproducibility of the results obtained among laboratories. SOPs gain special interest when the aim is the analysis of potentially unstable compounds. An SOP for analysis of lipid hydroperoxides (HpETEs) is here reported after optimization of the critical steps to be considered in their analysis in human serum from sampling to final analysis. The method is based on automated hyphenation between solid-phase extraction (SPE) and liquid chromatography-mass spectrometry (LC-MS). The developed research involves: (i) optimization of the SPE and LC-MS steps with a proper synchronization; (ii) validation of the method-viz. accuracy study (estimated as 86.4% as minimum value), evaluation of sensitivity and precision, which ranged from 2.5 to 7.0 ng/mL (0.25-0.70 ng on column) as quantification limit and precision below 13.2%), and robustness study (reusability of the cartridge for 5 times without affecting the accuracy and precision of the method); (iii) stability study, involving freeze-thaw stability, short-term and long-term stability and stock solution stability tests. The results thus obtained allow minimizing both random and systematic variation of the metabolic profiles of the target compounds by correct application of the established protocol.     

Influence of oleic acid on the structure of a mixture of hydrated model stratum corneum fatty acids and their soaps

The phase behaviour of a hydrated (32% (w/w) water) mixture of saturated and unsaturated fatty acids and their soaps has been examined. By progressively increasing the proportion of oleic acid, it could be shown that this unsaturated component exists partly as a separate phase within the others. The unsaturated fatty acids form an H_II phase at 25°C, whereas the saturated fatty acids form a fatty acid—soap crystal under the conditions employed. The results help to explain the role played by the fatty acids (and also cholesterol) within the lipid fraction of human stratum corneum. A feature of this study is a comparison of the results obtained by X-ray diffraction, differential scanning calorimetry and Fourier transform infrared spectroscopy.     

Raman Scattering from Liquid Crystals

Abstract

The use of Raman scattering technique as a tool for studying local order in complex liquid crystal systems is illustrated by three examples, namely the locally anisotropic liquid L-phase, lipid membranes, and PDLC's.