High-throughput screening assay for the quantification of Cer d18:1/16:0, d18:1/24:0, d18:1/24:1, d18:1/18:0, d18:1/14:0, d18:1/20:0, and d18:1/22:0 in HepG2 cells using RapidFire mass spectrometry.
Biomed Chromatogr
; 34(5): e4790, 2020 May.
Article
em En
| MEDLINE
| ID: mdl-31883352
ABSTRACT
Ceramides are known to be involved in various biological processes with their physiological levels elevated in various disease conditions such as diabetes, Alzheimer's, atherosclerosis. To facilitate the rapid screening of Cer d181/160, d181/240, d181/241, d181/180, d181/140, d181/200, and d181/220 inhibition in HepG2 cells, a RapidFire coupled to tandem mass spectrometry (RF-MS/MS) method has been developed. The RF platform provides an automated solid-phase extraction system that gave a throughput of 12.6 s per sample to an MS/MS system using electrospray ionization under the positive ion mode. Chromatographic separation of Cer d181/160, d181/240, d181/241, d181/180, d181/140, d181/200, and d181/220 was achieved using a ternary gradient on C8 type E cartridge. The MS/MS ion transitions monitored were 538.2 â 264.2, 650.7 â 264.2, 648.6 â 264.2, 566.4 â 264.2, 510.4 â 264.2, 594.4 â 264.2, 622.5 â 264.2, and 552.3 â 250.2 for Cer d181/160, d181/240, d181/241, d181/180, d181/140, d181/200, d181/220, and the internal standard (Cer d171/180), respectively. The RF-MS/MS methodology showed an excellent performance with an average Z' value of 0.5-0.7. This is the first report of an RF-MS/MS assay for screening of ceramides which is amenable for high-throughput screening.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Ceramidas
/
Espectrometria de Massas em Tandem
/
Ensaios de Triagem em Larga Escala
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
/
Screening_studies
Limite:
Humans
Idioma:
En
Revista:
Biomed Chromatogr
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Índia