Lipid structural information from a single equine embryo by MALDI-MS / Informação estrutural de lipídios a partir de um único embrião equino pela técnica de MALDI-MS

It was reported the potential of MALDI-MS for the characterization of lipid species present in a single equine embryo, and studied some lipid structures detected by collision induced dissociation (CID) experiments. In the positive ion mode spectrum, it were observed mostly protonated and sodiated species of sphingomyelins (SM), phosphatidylcholines (PC) and triacylglycerols (TAG). In the negative ion mode, it were observed phosphatidylethanolamines (PE) and phosphatidylinositols (PI). MS/MS spectrum of most intense lipid ions was performed to show MALDI-MS/MS structural information potential. MS/MS spectrum in the positive mode of m/z 760.6 (attributed as PC34:1) depicted characteristic PC fragments of m/z 184.1 (choline polar head), and the neutral loss (NL) of 183 (phosphorylcholine). For the ion of m/z 766.6 (attributed as PE 38:5), we observed the NL of 140, characteristic of PE. For the ion of m/z 808.7 (attributed as PC 38.5), besides the fragment at m/z 184.1 at the NL of 183, it was possible to observe the loss of trimethylamine (ion of m/z 749.6), and the cyclophosphane (ion of m/z 147.0). Finally, for the negative ion mode, we isolated and fragmented the ion at m/z 863.6, which was attributed as PI 36:1 due to the presence of m/z 153 (glycerol phosphate H2 O-H), 223 (phospho inositol 2H2 O-H), 241 (phospho inositol H2 O-H), 281 (oleic acid), and 581.3 (lysophosphoinositol H2 O-H). It was concluded that MALDI-MS allowed the detection of a broad range of PC, SM, PE, PI and TAG lipid species, as well as a fast and confident characterization of lipid structures from a single equine embryo.(AU)

É relatado o potencial da técnica de MALDI-MS para caracterizar espécies de lipídios presentes em um único embrião equino e estudadas algumas estruturas lipídicas detectadas por dissociação induzida por colisão (CID). No espectro de modo íon positivo, foram observadas espécies, principalmente, protonadas e sodiadas de esfingomielinas (SM), fosfatidileolinas (PC) e triacilgliceróis (TAG). No modo negativo, foram observadas fosfatidiletanolaminas (PE) e fosfatidilinositos (PI). Espectros de íons de lípidos com maior intensidade foram utilizados para demonstrar o potencial da informação estrutural por MALDI-MS/MS. O espectro no modo positivo de m/z (massa sobre carga) 760,6 (atribuída como PC34:1) apresentou características de fragmentos PC de m/z 184,1 (denominada cabeça polar de colina), além de perda neutral (NL) de m/z 183 (fosforilcolina). Para o íon de m/z 766,6 (atribuída como PE38:5), observou-se a NL de 140, característica do PE. Para o íon de m/z 808,7 (38,5 atribuído como PC), além do fragmento m/z 184,1 na NL de 183, foi possível observar a perda de trimetilamina (íon de m/z 749,6) e o ciclofosfano (íon de m/z 147,0). Finalmente, para o modo de íon negativo, foram isolados e fragmentados o íon de m/z 863,6 que foi atribuído como PI36:1, devido à presença de m/z 153 (fosfato de glicerol H2 O-H ), 223 (inositol fosfo - 2H2 O-H) , 241 (fosfoinositol H2 O-H), 281 (ácido oleico) e 581,3 (lisofosfoinositol H2 O+H). Foi concluído que a MALDI - MS permite a detecção de uma ampla gama de espécies de PC, SM, PE, PI e TAG lipídicas, bem como a caracterização rápida e confiante de estruturas lipídicas a partir de um único embrião equino.(AU)

Proteomics, metabolomics and lipidomics in reproductive biotechnologies: the MS solutions

Background : A broader view of living systems complexity is bringing important contributions to biological sciences, since the genome expression is affected by other classes of molecules, which in their turn interact themselves in cellular metabolic pathways and biochemical networks. This level of information has been made possible by the emergence of the omic strategies, such as proteomics, metabolomics and lipidomics, that are mainly based on mass spectrometry (MS) platforms. MS has presented an incredible development over the last years, evolving to a powerful and universal analytical technique. Its ability to analyze proteins and small molecules such as lipids, sugars and metabolites at the structural level, with sensitivity and speed inconceivable a few years ago, is the major driving force in the omic fields. The development of electrospray and matrix-assisted laser desorption/ionization (MALDI) ionization techniques has decisively contributed to the many applications of this technology nowadays. Herein, we present and discuss omic concepts and strategies, as well as detail basic principles of MS. Applications and future perspectives of these approaches are focused in the reproductive medicine area. Review: The omic technologies propose global characterization of specific classes of target biomolecules of cellular systems as a strategy to achieve comprehensive understanding of biological functions. The genomics, aimed at performing the entire genetic sequencing of organisms, represented the seminal step towards the understanding of the complex logic that orchestrates the function of all organisms or the defects leading to diseases. But to express the phenotype, information needs to flow from DNA via carrier biomolecules through processes that are being addressed by new omic sciences such as the transcriptomics, proteomics, metabolomics, glycomics, lipidomics, and fluxomics. Mass spectrometry (MS) is nowadays the most powerful technique for the structural characterization of biomolecules, and has therefore become the central technique for the omic sciences. Using revolutionary ionization techniques such as electrospray (ESI) and matrix-assisted laser desorption ionization (MALDI), a wide range of biomolecules such as peptides, proteins, lipids and sugars are efficiently transferred in intact ionized forms to the gas phase for MS analysis. The development of ESI-MS and MALDI-MS has been awarded the Nobel Prize for Chemistry in 2002, rocketing the application of MS in the omic sciences. More recently, ambient ionization MS techniques, such as desorption electrospray ionization (DESI) and easy ambient sonic-spray ionization (EASI), have been developed for ionization in the open atmosphere, in a workup free and high throughput fashion directly from sample in their original environments. For the more complex samples, the coupling with separation techniques such as liquid chromatrography (LC) as well as the use of tandem MS (LC-MS/MS) has allowed comprehensive mixture characterization of major biomolecules. Conclusion: This manuscript describes recent advances of MS in the proteomics, metabolomics and lipidomics for biological sciences, and points out the relevant contributions that MS is likely to bring to fundamental and applied research in human and animal embryo biotechnologies.