Double Cyclization Tandem Mass for Identification and Quantification of Phosphatidylcholines Using Isobaric Six-Plex Capillary nLC-MS/MS.
J Am Soc Mass Spectrom
; 35(7): 1403-1412, 2024 Jul 03.
Article
en En
| MEDLINE
| ID: mdl-38870035
ABSTRACT
Multiplexing of phosphatidylcholine analysis is hindered by a lack of appropriate derivatization. Presented here is a tagging scheme that uses a quaternary amine tag and targets the hydroxy group of the phosphate, which switches the net charge from neutral to +2. Quantitative yields were achieved from >99% reaction completion derived by dimethoxymethyl morpholinium (DMTMM) activation. Fragmentation of phosphatidylcholines (PCs) and lysophosphatidylcholines (LPCs) releases two trimethylamines and the acyl chains through neutral loss and generates a unique double cyclization constant mass reporter. Selective incorporation of isotopes onto the tag produces a six-plex set of isobaric reagents. For equivalent six-plex-labeled samples, <14% RSD was achieved, followed by a dynamic range of 110 without signal compression. Quantification of PCs/LPCs in human hepatic cancer cells was conducted as six-plex using data-dependent analysis tandem MS. We report a six-plex qualitative and quantitative isobaric tagging strategy expanding the limits of analyzing PCs/LPCs.
Texto completo:
1
Colección:
01-internacional
Asunto principal:
Fosfatidilcolinas
/
Espectrometría de Masas en Tándem
Límite:
Humans
Idioma:
En
Revista:
J Am Soc Mass Spectrom
Año:
2024
Tipo del documento:
Article
País de afiliación:
Estados Unidos