RESUMO
Cerebrosides, including glucosylceramides (GlcCers) and galactosylceramides (GalCers), are important membrane components of animal cells with deficiencies resulting in devastating lysosomal storage disorders. Their quantification is essential for disease diagnosis and a better understanding of disease mechanisms. The simultaneous quantification of GlcCer and GalCer isomers is, however, particularly challenging due to their virtually identical structures. To address this challenge, we developed a new LC/MS-based method using differential ion mobility spectrometry (DMS) capable of rapidly and reproducibly separating and quantifying isomeric cerebrosides in a single run. We show that this LC/ESI/DMS/MS/MS method exhibits robust quantitative performance within an analyte concentration range of 2.8-355 nM. We further report the simultaneous quantification of nine GlcCers (16:0, 18:0, 20:0, 22:0, 23:0, 24:1, 24:0, 25:0, and 26:0) and five GalCers (16:0, 22:0, 23:0, 24:1, and 24:0) molecular species in human plasma, as well as six GalCers (18:0, 22:0, 23:0, 24:1, 24:0 and 25:0) and two GlcCers (24:1 and 24:0) in human cerebrospinal fluid. Our method expands the potential of DMS technology in the field of glycosphingolipid analysis for both biomarker discovery and drug screening by enabling the unambiguous assignment and quantification of cerebroside lipid species in biological samples.
Assuntos
Cerebrosídeos/química , Cerebrosídeos/isolamento & purificação , Cromatografia Líquida/métodos , Espectrometria de Mobilidade Iônica/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos , Cerebrosídeos/sangue , Cerebrosídeos/líquido cefalorraquidiano , Cromatografia Líquida/normas , Feminino , Humanos , Espectrometria de Mobilidade Iônica/normas , Isomerismo , Pessoa de Meia-Idade , Padrões de Referência , Espectrometria de Massas por Ionização por Electrospray/normas , Espectrometria de Massas em Tandem/normas , Fatores de TempoRESUMO
A molecule isolated from the cerebrospinal fluid of sleep-deprived cats has been chemically characterized and identified as cis-9,10-octadecenoamide. Other fatty acid primary amides in addition to cis-9,10-octadecenoamide were identified as natural constituents of the cerebrospinal fluid of cat, rat, and human, indicating that these compounds compose a distinct family of brain lipids. Synthetic cis-9,10-octadecenoamide induced physiological sleep when injected into rats. Together, these results suggest that fatty acid primary amides may represent a previously unrecognized class of biological signaling molecules.
Assuntos
Química Encefálica , Cerebrosídeos/líquido cefalorraquidiano , Lipídeos/líquido cefalorraquidiano , Ácidos Oleicos/líquido cefalorraquidiano , Sono , Animais , Gatos , Cerebrosídeos/química , Cerebrosídeos/farmacologia , Humanos , Lipídeos/química , Lipídeos/farmacologia , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Peso Molecular , Ácidos Oleicos/química , Ácidos Oleicos/farmacologia , Ratos , Transdução de Sinais , Sono/efeitos dos fármacos , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Espectrofotometria InfravermelhoRESUMO
The author reports of the achieved results in a quantitative study of general lipids, phospholipids and cerebrosides in the CSF of 37 patients with demyelinating diseases, of 11 patients with vascular brain pathology and 7 with Van Bogart's panencephalitis. The control group consisted of 14 patients without focal lesions of the nervous system, with normal CSF indices. In demyelinating diseases there was a significant increase in the content of kephalines and cerebrosides. In Van Bogart's panencephalitis there was a much higher increase of kephalines, general lipids and phospholipids. In vascular brain disorders there was a moderate increase of all lipids. The possible pathochemical mechanisms of the depicted changes in the content of the lipids in the CSF are discussed.
Assuntos
Doenças Desmielinizantes/líquido cefalorraquidiano , Lipídeos/líquido cefalorraquidiano , Adolescente , Adulto , Cerebrosídeos/líquido cefalorraquidiano , Proteínas do Líquido Cefalorraquidiano/líquido cefalorraquidiano , Transtornos Cerebrovasculares/líquido cefalorraquidiano , Esclerose Cerebral Difusa de Schilder/líquido cefalorraquidiano , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/líquido cefalorraquidiano , Fosfatidiletanolaminas/líquido cefalorraquidiano , Fosfolipídeos/líquido cefalorraquidiano , Panencefalite Esclerosante Subaguda/líquido cefalorraquidianoAssuntos
Lipídeos/líquido cefalorraquidiano , Microquímica , Esclerose Lateral Amiotrófica/líquido cefalorraquidiano , Cerebrosídeos/líquido cefalorraquidiano , Colesterol/líquido cefalorraquidiano , Cromatografia em Camada Fina , Estudos de Avaliação como Assunto , Ácidos Graxos não Esterificados/líquido cefalorraquidiano , Humanos , Hidrocarbonetos/líquido cefalorraquidiano , Métodos , Fosfolipídeos/líquido cefalorraquidiano , Triglicerídeos/líquido cefalorraquidianoAssuntos
Neoplasias Encefálicas/líquido cefalorraquidiano , Glicolipídeos/líquido cefalorraquidiano , Metástase Neoplásica/líquido cefalorraquidiano , Fosfolipídeos/líquido cefalorraquidiano , Neoplasias da Mama/líquido cefalorraquidiano , Cerebrosídeos/líquido cefalorraquidiano , Cromatografia em Camada Fina , Gangliosídeos/líquido cefalorraquidiano , Humanos , Neoplasias Pulmonares/líquido cefalorraquidiano , Métodos , Fosfatidiletanolaminas/líquido cefalorraquidiano , Punção EspinalRESUMO
A technique was developed to isolate sufficient material for compositional analysis of cerebroside from pooled human cerebrospinal fluid. The carbohydrate moiety was principally galactose. The sphingosine base and fatty acid compositions were found to be similar to that of brain cerebroside. The presence of a contaminant in commercial silica gel which chromatographed like the trimethylsilyl derivative of glucose is described.
