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1.
Food Chem ; 334: 127558, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32711269

RESUMO

Shellfishes contain plasmalogens correlating to the functions of brain, heart, etc. Herein, a mild acid hydrolysis and hydrophilic interaction chromatography (HILIC) tandem mass spectrometry method was developed for analyzing plasmalogens in six shellfish species. A total of 19 plasmalogen molecular species were successfully identified, including nine phosphatidylcholine plasmalogen (plasPC), seven phosphatidylethanolamine plasmalogen (plasPE), and three phosphatidylserine plasmalogen (plasPS). The quantitative results indicated that mussel (32 µg·mg-1) possessed the highest content of plasmalogens, followed by oyster (21 µg·mg-1) and razor clam (15 µg·mg-1). The statistic models showed that the plasPE P-18:0/20:5 (m/z 748), plasPE P-16:0/22:2 & P-18:0/20:2 (m/z 754) and plasPS were the most contributing difference between shellfishes. The results indicated that this method was sensitive and precise to determine plasmalogens in shellfish, and mussel was demonstrated to be a good choice for the large-scale preparation of plasmalogens.


Assuntos
Bivalves/química , Cromatografia/métodos , Plasmalogênios/análise , Frutos do Mar/análise , Espectrometria de Massas em Tandem/métodos , Animais , Análise de Alimentos/métodos , Interações Hidrofóbicas e Hidrofílicas , Lipidômica/métodos , Ostrea/química , Fosfatidilcolinas/análise , Fosfatidilserinas/análise , Plasmalogênios/química
2.
Food Chem ; 339: 128114, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33152890

RESUMO

Lipids are hydrophobic metabolites implicated in tea flavor quality. Understanding their transformations during tea manufacture is of particular interest. To date, the detailed lipid composition and variations during green tea manufacture are largely unknown. Herein, we performed a comprehensive characterization of the dynamic changes of lipids during green tea manufacture, by applying nontargeted lipidomics using ultrahigh performance liquid chromatography-quadrupole-Orbitrap mass spectrometry (UHPLC-Q-Exactive/MS) combined with chemometric tools. Totally, 283 lipid species were detected, covering 20 subclasses. Significant lipidomic variations were observed during green tea manufacture, especially in the fixation stage, mainly associated with chlorophyll decomposition, phosphatidic acids (PAs) reduction and glycolipids degradation, which potentially contribute to tea color and aroma quality. Specifically, the most prominent decrease of PAs content during green tea manufacture was identified for the first time. This study provides insights into the lipid metabolic fates upon green tea manufacture, and their roles in green tea sensory quality.


Assuntos
Lipidômica/métodos , Lipídeos/análise , Lipídeos/química , Chá/química , China , Cromatografia Líquida de Alta Pressão , Cor , Indústria de Processamento de Alimentos , Metabolismo dos Lipídeos , Espectrometria de Massas , Odorantes/análise
3.
J Am Soc Mass Spectrom ; 31(10): 2013-2024, 2020 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-32880453

RESUMO

As corona virus disease 2019 (COVID-19) is a rapidly growing public health crisis across the world, our knowledge of meaningful diagnostic tests and treatment for severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) is still evolving. This novel coronavirus disease COVID-19 can be diagnosed using RT-PCR, but inadequate access to reagents, equipment, and a nonspecific target has slowed disease detection and management. Precision medicine, individualized patient care, requires suitable diagnostics approaches to tackle the challenging aspects of viral outbreaks where many tests are needed in a rapid and deployable approach. Mass spectrometry (MS)-based technologies such as proteomics, glycomics, lipidomics, and metabolomics have been applied in disease outbreaks for identification of infectious disease agents such as virus and bacteria and the molecular phenomena associated with pathogenesis. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS) is widely used in clinical diagnostics in the United States and Europe for bacterial pathogen identification. Paper spray ionization mass spectrometry (PSI-MS), a rapid ambient MS technique, has recently open a new opportunity for future clinical investigation to diagnose pathogens. Ultra-high-pressure liquid chromatography coupled high-resolution mass spectrometry (UHPLC-HRMS)-based metabolomics and lipidomics have been employed in large-scale biomedical research to discriminate infectious pathogens and uncover biomarkers associated with pathogenesis. PCR-MS has emerged as a new technology with the capability to directly identify known pathogens from the clinical specimens and the potential to identify genetic evidence of undiscovered pathogens. Moreover, miniaturized MS offers possible applications with relatively fast, highly sensitive, and potentially portable ways to analyze for viral compounds. However, beneficial aspects of these rapidly growing MS technologies in pandemics like COVID-19 outbreaks has been limited. Hence, this perspective gives a brief of the existing knowledge, current challenges, and opportunities for MS-based techniques as a promising avenue in studying emerging pathogen outbreaks such as COVID-19.


Assuntos
Infecções por Coronavirus/etiologia , Lipidômica/métodos , Espectrometria de Massas/métodos , Metabolômica/métodos , Pneumonia Viral/etiologia , Proteômica/métodos , Cromatografia Líquida de Alta Pressão , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Glicômica/métodos , Humanos , Pandemias , Reação em Cadeia da Polimerase , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
4.
Life Sci ; 260: 118489, 2020 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-32976882

RESUMO

AIMS: Cervical cancer (CC) is a common tumor of women worldwide. Here, we conducted a non-targeted lipidomic study to discover novel lipid biomarkers for early-stage CC. MAIN METHODS: The lipidomic analysis of 71 samples in discovery set and 72 samples in validation set were performed by coupling ultra-high-pressure liquid chromatography (UHPLC) with quadrupole time-of-flight tandem mass spectrometry (Q-TOF-MS). Lipids with variable importance (VIP) values greater than 1, adj. p < 0.05 (the adjusted p value obtained from false discovery rate correction) and fold change (FC) higher than 1.5 were reserved as potential biomarkers. Subsequently, receiver operating characteristic (ROC) curve and binary logistic regression were implemented to assess the diagnostic potential of these biomarkers and to acquire the best biomarker combination. KEY FINDINGS: A lipid biomarker panel, including phosphatidylcholine (PC, PC 14:0/18:2) and phosphatidylethanolamine (PE, PE 15:1e/22:6 and PE 16:1e/18:2), was established. This panel was effective in distinguishing between CC and non-CC (squamous intraepithelial lesions [SIL] and healthy controls) within the area under the ROC curve (AUC), sensitivity, and specificity reaching 0.966, 0.952, and 0.860 for discovery set and 0.961, 0.920, and 0.915 for external validation set. Furthermore, this panel was also capable of discriminating early-stage CC from SIL with AUC, sensitivity, and specificity reaching 0.946, 0.952, and 0.800 for discovery set and 0.956, 0.960, and 0.815 for external validation set. SIGNIFICANCE: The combination of PC 14:0/18:2, PE 15:1e/22:6, and PE 16:1e/18:2 could serve as a promising serum biomarker for discriminating early-stage CC from SIL and healthy subjects.


Assuntos
Biomarcadores Tumorais/sangue , Lipidômica/métodos , Lipídeos/sangue , Neoplasias do Colo do Útero/sangue , Adulto , Estudos de Casos e Controles , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Espectrometria de Massas/métodos , Pessoa de Meia-Idade , Análise Multivariada , Fosfatidilcolinas/sangue , Fosfatidiletanolaminas/sangue , Reprodutibilidade dos Testes , Lesões Intraepiteliais Escamosas/sangue , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/patologia
5.
PLoS One ; 15(9): e0238921, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32925944

RESUMO

The objective of this study was to characterize the chemical composition and lipid profile of colostrum and milk of purebred Quarter Horse mares. Thirty-four (34) purebred mares were selected, which were then separated into groups according to age, birth order and lactation stage. Colostrum samples were collected in the first six hours after delivery and milk samples from the 7th postpartum day, with intervals of 14 days until the end of lactation. The samples were refrigerated and sent to the Milk Laboratory of the University (Laboleite-UFRN), where they were analyzed for chemical composition. Colostrum was assessed by refractometry. The lipid profile was determined by gas chromatography through a separation of methyl esters. The data were tabulated and subjected to descriptive statistics and analysis of variance by the F-Test, and the groups were compared by the Tukey test using a significance level of 5%. There was high protein content and reduced lactose content for the colostrum of the Quarter Horse mares, differing from other breeds. The milk composition was not influenced by the mares' age. However, variations in the lactation stage and in the birth order of the Quarter Horse mares altered the milk's chemical composition. There is variation in the lipid composition of milk according to the lactation stage, without changing the characteristic profile of the mares' milk or diminishing the nutritional quality of the lipid fraction.


Assuntos
Colostro/química , Lipidômica/métodos , Lipídeos/análise , Leite/química , Animais , Peso Corporal , Cruzamento , Cromatografia Gasosa , Feminino , Cavalos , Valor Nutritivo , Gravidez , Refratometria
6.
Nat Commun ; 11(1): 4107, 2020 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-32796836

RESUMO

Foamy macrophages, which have prominent lipid droplets (LDs), are found in a variety of disease states. Toll-like receptor agonists drive triacylglycerol (TG)-rich LD development in macrophages. Here we explore the basis and significance of this process. Our findings indicate that LD development is the result of metabolic commitment to TG synthesis on a background of decreased fatty acid oxidation. TG synthesis is essential for optimal inflammatory macrophage activation as its inhibition, which prevents LD development, has marked effects on the production of inflammatory mediators, including IL-1ß, IL-6 and PGE2, and on phagocytic capacity. The failure of inflammatory macrophages to make PGE2 when TG-synthesis is inhibited is critical for this phenotype, as addition of exogenous PGE2 is able to reverse the anti-inflammatory effects of TG synthesis inhibition. These findings place LDs in a position of central importance in inflammatory macrophage activation.


Assuntos
Inflamação/metabolismo , Lipidômica/métodos , Triglicerídeos/metabolismo , Animais , Células Cultivadas , Citometria de Fluxo , Glucose/metabolismo , Glicerol/metabolismo , Células HEK293 , Humanos , Metabolismo dos Lipídeos/fisiologia , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Confocal , Microscopia Eletrônica , Palmitatos/metabolismo , Análise de Sequência de RNA
7.
J Vis Exp ; (159)2020 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-32538912

RESUMO

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still in need. In recent years, mass spectrometry has become a method of choice for untargeted profiling of discriminatory compound as potential biomarkers of a disease. Biofluids are generally considered as preferable matrices given their accessibility and easier sample processing while direct tissue profiling provides more selective information about a given cancer. Preparation of tissues for the analysis via traditional methods is much more complex and time-consuming, and, therefore, not suitable for fast on-site analysis. The current work presents a protocol combining sample preparation and extraction of small molecules on-site, immediately after tumor resection. The sampling device, which is of the size of an acupuncture needle, can be inserted directly into the tissue and then transported to the nearby laboratory for instrumental analysis. The results of metabolomics and lipidomics analyses demonstrate the capability of the approach for the establishment of phenotypes of tumors related to the histological origin of the tumor, malignancy, and genetic mutations, as well as for the selection of discriminating compounds or potential biomarkers. The non-destructive nature of the technique permits subsequent performance of routinely used tests e.g., histological tests, on the same samples used for SPME analysis, thus enabling attainment of more comprehensive information to support personalized diagnostics.


Assuntos
Neoplasias Encefálicas/metabolismo , Lipidômica/métodos , Metabolômica/métodos , Biomarcadores/metabolismo , Humanos , Espectrometria de Massas , Manejo de Espécimes
8.
Nat Commun ; 11(1): 2967, 2020 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-32528049

RESUMO

The recent outbreak of Zika virus (ZIKV) was associated with birth defects and pregnancy loss when maternal infection occurs in early pregnancy, but specific mechanisms driving placental insufficiency and subsequent ZIKV-mediated pathogenesis remain unclear. Here we show, using large scale metabolomics, that ZIKV infection reprograms placental lipidome by impairing the lipogenesis pathways. ZIKV-induced metabolic alterations provide building blocks for lipid droplet biogenesis and intracellular membrane rearrangements to support viral replication. Furthermore, lipidome reprogramming by ZIKV is paralleled by the mitochondrial dysfunction and inflammatory immune imbalance, which contribute to placental damage. In addition, we demonstrate the efficacy of a commercially available inhibitor in limiting ZIKV infection, provides a proof-of-concept for blocking congenital infection by targeting metabolic pathways. Collectively, our study provides mechanistic insights on how ZIKV targets essential hubs of the lipid metabolism that may lead to placental dysfunction and loss of barrier function.


Assuntos
Placenta/virologia , Infecção por Zika virus/imunologia , Infecção por Zika virus/metabolismo , Feminino , Humanos , Transmissão Vertical de Doença Infecciosa , Lipidômica/métodos , Gravidez , Complicações Infecciosas na Gravidez/imunologia , Complicações Infecciosas na Gravidez/metabolismo , Zika virus/imunologia , Zika virus/patogenicidade
9.
Nat Commun ; 11(1): 3186, 2020 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-32581242

RESUMO

Mass spectrometry based metabolomics is a widely used approach in biomedical research. However, current methods coupling mass spectrometry with chromatography are time-consuming and not suitable for high-throughput analysis of thousands of samples. An alternative approach is flow-injection mass spectrometry (FI-MS) in which samples are directly injected to the ionization source. Here, we show that the sensitivity of Orbitrap FI-MS metabolomics methods is limited by ion competition effect. We describe an approach for overcoming this effect by analyzing the distribution of ion m/z values and computationally determining a series of optimal scan ranges. This enables reproducible detection of ~9,000 and ~10,000 m/z features in metabolomics and lipidomics analysis of serum samples, respectively, with a sample scan time of ~15 s and duty time of ~30 s; a ~50% increase versus current spectral-stitching FI-MS. This approach facilitates high-throughput metabolomics for a variety of applications, including biomarker discovery and functional genomics screens.


Assuntos
Análise de Injeção de Fluxo/métodos , Espectrometria de Massas/métodos , Metabolômica/métodos , Linhagem Celular Tumoral , Análise de Injeção de Fluxo/normas , Ensaios de Triagem em Larga Escala , Humanos , Íons/química , Lipidômica/métodos , Espectrometria de Massas/normas , Metabolômica/normas , Soro/química , Soro/metabolismo
10.
PLoS Negl Trop Dis ; 14(5): e0008145, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32401760

RESUMO

Schistosomes are human pathogens causing the neglected tropical disease schistosomiasis, which occurs worldwide in (sub-)tropical regions. This infectious disease is often associated with poverty, and more than 700 million people are at risk of infection. Exploitation of novel habitats and limited therapeutic options brought schistosomes into research focus. Schistosomes are the only trematodes that have evolved separate sexes. They are covered by their metabolically active tegument, a surface area representing the interface between male and female in their permanent mating contact but also between parasite and host. The tegument comprises, besides others, numerous specific lipid compounds. Limited information is available on the exact lipid composition and its spatial distribution. We used atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization (AP-SMALDI) mass spectrometry imaging (MSI) to characterize the Schistosoma mansoni tegument surface in comparison to tissue sections of whole worms or couples. We found that phosphatidylcholines (PC) and specific phosphatidylethanolamines (PE) are significantly more abundant inside the worm body compared to the tegument. On the other hand, the latter was found to be enriched in sphingomyelins (SM), phosphatidylserines (PS), lysophosphatidylcholines (LPC), and specific PE species. We further investigated lipid classes concerning number of carbon atoms in fatty acyl chains as well as the degree of unsaturation and found pronounced differences between the tegument and whole-worm body. Furthermore, differences between male and female teguments were found. The lipid composition of S. mansoni tissues has been investigated in an untargeted, spatially resolved manner for the first time.


Assuntos
Lipidômica/métodos , Lipídeos/química , Schistosoma mansoni/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Feminino , Metabolismo dos Lipídeos , Masculino , Schistosoma mansoni/metabolismo
11.
Arch Biochem Biophys ; 688: 108431, 2020 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-32461102

RESUMO

Phenylketonuria (PKU) is the most prevalent inborn error of amino acid metabolism. The disease is due to the deficiency of phenylalanine (Phe) hydroxylase activity, which causes the accumulation of Phe. Early diagnosis through neonatal screening is essential for early treatment implementation, avoiding cognitive impairment and other irreversible sequelae. Treatment is based on Phe restriction in the diet that should be maintained throughout life. High dietary restrictions can lead to imbalances in specific nutrients, notably lipids. Previous studies in PKU patients revealed changes in levels of plasma/serum lipoprotein lipids, as well as in fatty acid profile of plasma and red blood cells. Most studies showed a decrease in important polyunsaturated fatty acids, namely DHA (22:6n-3), AA (20:4n-6) and EPA (20:5n-6). Increased oxidative stress and subsequent lipid peroxidation have also been observed in PKU. Despite the evidences that the lipid profile is changed in PKU patients, more studies are needed to understand in detail how lipidome is affected. As highlighted in this review, mass spectrometry-based lipidomics is a promising approach to evaluate the effect of the diet restrictions on lipid metabolism in PKU patients, monitor their outcome, namely concerning the risk for other chronic diseases, and find possible prognosis biomarkers.


Assuntos
Ácidos Graxos/metabolismo , Lipidômica , Lipoproteínas/metabolismo , Fenilcetonúrias/fisiopatologia , Ácidos Graxos/análise , Humanos , Inflamação/complicações , Inflamação/fisiopatologia , Peroxidação de Lipídeos/fisiologia , Lipidômica/métodos , Lipoproteínas/análise , Estresse Oxidativo/fisiologia , Fenilcetonúrias/complicações , Fenilcetonúrias/dietoterapia , Triglicerídeos/análise , Triglicerídeos/metabolismo
12.
Nat Commun ; 11(1): 2057, 2020 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-32345972

RESUMO

Mass spectrometry (MS)-based targeted lipidomics enables the robust quantification of selected lipids under various biological conditions but comprehensive software tools to support such analyses are lacking. Here we present LipidCreator, a software that fully supports targeted lipidomics assay development. LipidCreator offers a comprehensive framework to compute MS/MS fragment masses for over 60 lipid classes. LipidCreator provides all functionalities needed to define fragments, manage stable isotope labeling, optimize collision energy and generate in silico spectral libraries. We validate LipidCreator assays computationally and analytically and prove that it is capable to generate large targeted experiments to analyze blood and to dissect lipid-signaling pathways such as in human platelets.


Assuntos
Lipidômica/métodos , Software , Adulto , Plaquetas/metabolismo , Calibragem , Feminino , Humanos , Lipídeos/sangue , Lipídeos/química , Masculino , Ativação Plaquetária , Probabilidade , Reprodutibilidade dos Testes , Transdução de Sinais , Adulto Jovem
13.
PLoS One ; 15(4): e0231289, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32287294

RESUMO

Breast cancer (BC) is the most prevalent type of cancer in women in western countries. BC mortality has not declined despite early detection by screening, indicating the need for better informed treatment decisions. Therefore, a novel noninvasive diagnostic tool for BC would give the opportunity of subtype-specific treatment and improved prospects for the patients. Heterogeneity of BC tumor subtypes is reflected in the expression levels of enzymes in lipid metabolism. The aim of the study was to investigate whether the subtype defined by the transcriptome is reflected in the lipidome of BC cell lines. A liquid chromatography mass spectrometry (LC-MS) platform was applied to analyze the lipidome of six cell lines derived from human BC cell lines representing different BC subtypes. We identified an increased abundance of triacylglycerols (TG) ≥ C-48 with moderate or multiple unsaturation in fatty acyl chains and down-regulated ether-phosphatidylethanolamines (PE) (C-34 to C-38) in cell lines representing estrogen receptor and progesterone receptor positive tumor subtypes. In a cell line representing HER2-overexpressing tumor subtype an elevated expression of TG (≤ C-46), phosphatidylcholines (PC) and PE containing short-chained (≤ C-16) saturated or monounsaturated fatty acids were observed. Increased abundance of PC ≥ C-40 was found in cell lines of triple negative BC subtype. In addition, differences were detected in lipidomes within these previously defined subtypes. We conclude that subtypes defined by the transcriptome are indeed reflected in differences in the lipidome and, furthermore, potentially biologically relevant differences may exist within these defined subtypes.


Assuntos
Neoplasias da Mama/classificação , Neoplasias da Mama/metabolismo , Metabolismo dos Lipídeos , Lipidômica/métodos , Linhagem Celular Tumoral , Feminino , Humanos , Fosfatidilcolinas/metabolismo , Fosfatidiletanolaminas/metabolismo , Triglicerídeos/metabolismo
14.
J Gastroenterol Hepatol ; 35(8): 1355-1364, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32285970

RESUMO

BACKGROUND AND AIM: Lipids play important roles in inflammation and may be involved in the pathophysiology of inflammatory bowel disease (IBD). Here, we evaluated the characteristics of the plasma lipid profile in patients with IBD. METHODS: Plasma samples were collected from 20 patients with Crohn's disease (CD), 20 patients with ulcerative colitis (UC), and 10 healthy volunteers (HVs) after overnight fasting. The subjects were men between 20 and 49 years of age with no history of hyperlipidemia. A total of 698 molecular species in 22 lipid classes were analyzed by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. RESULTS: Lipid classes of lysophosphatidic acid, lysophosphatidylserine (LPS), phosphatidylserine (PS), and shingosine-1-phosphate (S1P) were significantly increased in UC patients compared with the HV. The LPS, PS, and S1P levels were significantly increased, while those of lysophosphatidylinositol and phosphatidylcholine were significantly decreased in CD patients compared with HV. Among PS species, the levels of PSacyl (PSa) 40:3, PSa 38:3, and PSa 42:4 were significantly higher in CD patients, both active and remissive stage, than in HV. The LPS 18:0 level was significantly higher in CD and UC patients compared with HV. PSa 40:3 and PSa 38:3 levels positively correlated with the Crohn's Disease Activity Index, erythrocyte sedimentation rate, and platelet count and negatively correlated with hemoglobin, hematocrit, and albumin levels in CD patients. CONCLUSION: The lipid profile in IBD patients exhibits significant alterations, and PS levels are associated with clinical disease activity in CD patients.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Doenças Inflamatórias Intestinais/diagnóstico , Lipidômica/métodos , Fosfatidilserinas/sangue , Espectrometria de Massas por Ionização por Electrospray/métodos , Adulto , Biomarcadores/sangue , Colite Ulcerativa/sangue , Colite Ulcerativa/diagnóstico , Doença de Crohn/sangue , Doença de Crohn/diagnóstico , Feminino , Humanos , Doenças Inflamatórias Intestinais/sangue , Lisofosfolipídeos/sangue , Masculino , Pessoa de Meia-Idade , Adulto Jovem
15.
Food Chem ; 318: 126504, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32146310

RESUMO

Tuna adulteration and mislabeling are serious problem worldwide and have caused economic loss and consumer rights violation. In this study, an electrometric knife (iKnife) coupling rapid evaporative ionization mass spectrometry (REIMS) and a multivariate recognition model were developed and employed for in situ and real-time authentication of four tuna species without sample preparation. The results showed that the lipidomic profiles were successfully acquired and the differences in fatty acids and phospholipids were statistically analyzed to be significant (p < 0.05). The model displayed the superb classification accuracy (>93%) and validation (R2(Y) = 0.992, Q2 = 0.986), and the main contributors of m/z 817.64, m/z 809.68, etc. were screened out to be used as potential biomarkers. Based on this technique, the identity of blind tuna samples could be unambiguously authenticated with the results displayed on a monitor screen directly. This study provided a front-line rapid detection method to prove the authenticity of tuna species.


Assuntos
Contaminação de Alimentos/análise , Lipidômica/métodos , Espectrometria de Massas/métodos , Animais , Ácidos Graxos/química , Fosfolipídeos/química , Atum
16.
Sci Adv ; 6(12): eaay4361, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32219161

RESUMO

Biomaterials composed of extracellular matrix (ECM) provide both mechanical support and a reservoir of constructive signaling molecules that promote functional tissue repair. Recently, matrix-bound nanovesicles (MBVs) have been reported as an integral component of ECM bioscaffolds. Although liquid-phase extracellular vesicles (EVs) have been the subject of intense investigation, their similarity to MBV is limited to size and shape. Liquid chromatography-mass spectrometry (LC-MS)-based lipidomics and redox lipidomics were used to conduct a detailed comparison of liquid-phase EV and MBV phospholipids. Combined with comprehensive RNA sequencing and bioinformatic analysis of the intravesicular cargo, we show that MBVs are a distinct and unique subpopulation of EV and a distinguishing feature of ECM-based biomaterials. The results begin to identify the differential biologic activities mediated by EV that are secreted by tissue-resident cells and deposited within the ECM.


Assuntos
Vesículas Extracelulares , Lipidômica , Nanopartículas , Análise de Sequência de RNA , Células 3T3 , Animais , Materiais Biocompatíveis , Cromatografia Líquida , Matriz Extracelular , Ácidos Graxos/metabolismo , Lipidômica/métodos , Microextração em Fase Líquida , Camundongos , Fosfolipídeos/metabolismo , Frações Subcelulares
17.
Anal Bioanal Chem ; 412(10): 2365-2374, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32130438

RESUMO

In this work, a lipidomics workflow based on offline semi-preparative lipid class-specific fractionation by supercritical fluid chromatography (SFC) followed by high-resolution mass spectrometry was introduced. The powerful SFC approach offered separation of a wide polarity range for lipids, enabled enrichment (up to 3 orders of magnitude) of lipids, selective fractionation of 14 lipid classes/subclasses, and increased dynamic range enabling in-depth characterization. A significantly increased coverage of low abundant lipids improving lipid identification by numbers and degree (species and molecular level) was obtained in Pichia pastoris when comparing high-resolution mass spectrometry based lipidomics with and without prior fractionation. Proof-of-principle experiments using a standard reference material (SRM 1950, NIST) for human plasma showed that the proposed strategy enabled quantitative lipidomics. Indeed, for 70 lipids, the consensus values available for this sample could be met. Thus, the novel workflow is ideally suited for lipid class-specific purification/isolation from milligram amounts of sample while not compromising on omics type of analysis (identification and quantification). Finally, compared with established fractionation/pre-concentration approaches, semi-preparative SFC is superior in terms of versatility, as it involved only volatile modifiers and salt additives facilitating any follow-up use such as qualitative or quantitate analysis or further purification down to the single lipid species level. Graphical Abstract.


Assuntos
Cromatografia com Fluido Supercrítico/métodos , Lipidômica/métodos , Lipídeos/química , Espectrometria de Massas/métodos , Humanos , Metabolismo dos Lipídeos , Lipídeos/sangue , Pichia/química , Pichia/metabolismo , Plasma/química
18.
Molecules ; 25(3)2020 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-32023992

RESUMO

Ceramides are important lipid metabolites for primal skin functions. There is increasing evidence that alteration of the profile and metabolism of ceramides is associated with skin diseases, such as psoriasis vulgaris. Most studies have reported alteration in ceramide content in the stratum corneum, but these have been scarcely reported for other skin layers. In the present work, we aimed to explore changes in the ceramide profile of fibroblasts and keratinocytes in patients with psoriasis vulgaris and healthy subjects. Using the reversed-phase liquid chromatography-quadrupole-time-of-flight-tandem-mass spectrometry (RPLC-QTOF-MS/MS) platform, we identified ceramide containing non-hydroxy fatty acid ([N]), α-hydroxy fatty acid ([A]), and esterified ω-hydroxy fatty acid ([EO]) and 3 sphingoid bases, dihydrosphingosine ([DS]), sphingosine ([S]), and phytosphingosine ([P]). We found that in the keratinocytes of patients with psoriasis, CER[NS], CER[NP], CER[AS], CER[ADS], CER[AP] and CER[EOS] tended to be expressed at higher relative levels, whereas CER[NDS] tended to be expressed with lower levels than in healthy subjects. In the case of fibroblasts, significant differences were observed, mainly in the three ceramide classes (CER[AS], CER[ADS] and CER[EOS]), which were expressed at significantly higher levels in patients with psoriasis. The most significant alteration in the fibroblasts involved elevated levels of CER[EOS] that contained ester-linked fatty acids. Our findings provide insights into the ceramide profile in the dermis and epidermis of patients with psoriasis and contribute for the research in this field, focusing on the role of keratinocyte-fibroblast crosstalk in the development of psoriasis vulgaris.


Assuntos
Ceramidas/análise , Queratinócitos/química , Lipidômica/métodos , Psoríase/metabolismo , Adulto , Estudos de Casos e Controles , Ceramidas/classificação , Cromatografia de Fase Reversa , Derme/química , Epiderme/química , Feminino , Fibroblastos/química , Humanos , Masculino , Pessoa de Meia-Idade , Espectrometria de Massas em Tandem
19.
J Am Soc Mass Spectrom ; 31(2): 319-325, 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-32031399

RESUMO

Infrared matrix-assisted laser desorption ionization (IR-MALDESI) is an ambient mass spectrometry imaging (MSI) technique that relies on electrospray ionization (ESI) for ion generation of desorbed neutrals. Although many mechanisms in IR-MALDESI have been studied in depth, there has not yet been a comprehensive study of how the ESI parameters change the profiles of tissue specific lipids. Acetonitrile (ACN)/water and methanol (MeOH)/water solvent systems and compositions were varied across a series of applied ESI voltages during IR-MALDESI analysis of rat liver tissue. Gradients of 12 min were run from 5 to 95% organic solvent in both positive and negative polarities across 11 voltages between 2.25 and 4.5 kV. These experiments informed longer gradients (25-30 min) across shorter solvent gradient ranges with fewer voltages. Optimal ESI parameters for lipidomics were determined by the number and abundance of detected lipids and the relative proportion of background ions. In positive polarity, the best solvent composition was 60-75% ACN/40-25% H2O with 0.2% formic acid at 3.2 kV applied voltage. The best parameters for negative polarity analysis are 45-55% ACN/55-45% H2O with 1 mM of acetic acid for voltages between 2.25 and 3.2 kV. Using these defined parameters, IR-MALDESI positive polarity lipidomics studies can increase lipid abundances 3-fold, with 15% greater coverage, while an abundance increase of 1.5-fold and 10% more coverage can be achieved relative to commonly used parameters in negative polarity.


Assuntos
Lipidômica/métodos , Imagem Molecular/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Lipídeos/análise , Fígado/química , Fígado/diagnóstico por imagem , Ratos
20.
J Am Soc Mass Spectrom ; 31(2): 463-466, 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-32031403

RESUMO

Natural lipidomes represent a complex mixture of lipid molecular species with a variety of biological and signaling functions. Modern mass spectrometry (MS)-based analytical platforms are often used to resolve the complexity of natural lipidomes. The quantitative transfer of lipid molecular species in the gas phase during the electrospray ionization required for MS analysis might be challenged by lipid in-source fragmentation (ISF) hampering their accurate identification and quantification. Here we evaluated the effect of transmission radio frequency (RF) levels and ion transfer temperatures (ITTs) on the analysis of four different lipids (ceramide, cholesteryl ester, phosphatidylethanolamine, and triacylglyceride) ionized in positive ion mode on three different Orbitrap-based platforms. ITT and RF levels were ramped in a systematic way to determine the best settings, allowing the most sensitive detection accompanied by the lowest ISF of a lipid. The extent of the ISF was shown to depend on the configurations of the transmission devices (S-lens vs letterbox/ion funnel) at defined RF and ITT levels for each studied lipid class. We provide here the recommendations for reducing the extent of lipid ISF without a significant loss in sensitivity for Q Exactive HF, Q Exactive HF-X, and Orbitrap Fusion Lumos platforms.


Assuntos
Lipidômica/métodos , Lipídeos/análise , Lipídeos/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Ceramidas , Ésteres do Colesterol , Fosfatidiletanolaminas , Sensibilidade e Especificidade
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