Gas chromatography with mass spectrometry analysis of phosphoserine, phosphoethanolamine, phosphoglycerol, and phosphate

Abstract

A new, rapid, sensitive, robust, and reliable method has been developed for the qualitative analysis of phosphoserine, phosphoethanolamine, phosphoglycerol, and phosphate using gas chromatography with mass spectrometry and two-step trimethylsilylation. The method employs hexamethyldisilazane for silylation of the phosphate and hydroxyl groups in the first phase and bis(trimethylsilyl)trifluoroacetamide for silylation of the less-reactive amino groups in the second phase. This order is of key importance for the method because of the different reactivities of the two reagents and the mechanism of derivatization of the active groups of the analytes. Trimethylsilylated derivatives of the analytes were identified on the basis of their retention times and mass spectra. The probable structures of the major fragments were identified in the spectra of the trimethylsilylated derivatives and characteristic m/z fragments were selected for each analyte. Fragments with m/z 73 and 299 occurred in the spectra of all the analytes. The characteristic retention data were employed to calculate the retention indices of the individual silylated phosphorylated substances in the hydrocarbon range C12-C19 for the DB-5ms column. The method was employed to measure the polar fraction of the hydrolysate of the cytoplasmic membrane of Bacillus subtilis. The detection limits vary between 5 μg/mL (trimethylsilylated phosphate) and 72 μg/mL (trimethylsilylated phosphoethanolamine).

Simultaneous analysis of carbohydrates, polyols and amines in urine samples using chemical ionization gas chromatography with tandem mass spectrometry

“… and Bierhanzl et al. , with HMDS and BSTFA was applied for the trimethylsilylation of polar functional groups of the target analytes present in the samples. The procedure was as follows: (i) in the first step, 700 μL of HMDS + ACN mixture (1:1, v/v) was added as a silylation agent to 20 μL of urine sample for the derivatization of easily silylable functional groups (e.g.…”

“…The aim of this work was the development of a new method enabling the simultaneous derivatization of carbohydrates, polyols and amines in urine samples without any sample pretreatment before derivatization, based on a previously published method by the authors , and their subsequent analysis by GC–MS/MS with positive chemical ionization. The newly developed method was used for the analysis of the selected compounds in urine samples of healthy individuals and neonates with galactosemia.…”

The effects of molecular collisions between the mobile phase and the solute in gas-solid chromatography

“…This paper addresses these interactions using gas‐solid chromatography and provides a new perspective on the chromatographic process. Chromatography is an indispensable means of analysis and separation in many fields . This significance motivates research on the basic theory of chromatography.…”

Avaliação antitumoral da fosfoetanolamina sintética e da formulação lipossomal DODAC/fosfoetanolamina sintética em células tumorais de mama humana

“…Os FLs fazem parte da fração não polar de amostras biológicas e são investigados do ponto de vista dos teores de ácidos graxos em conjunto com lipídios apolares. PLs são formados por um esqueleto de glicerol com duas moléculas de éter ligado a ácidos graxos (parte hidrofóbica), o qual o grupo fosfato funcional está ligado, o que é frequentemente modificado por uma estrutura de baixo peso molecular adicional (parte hidrofílica, por exemplo, serina, etanolamina, glicerol, colina, etc) (BIERHANZL et al, 2015). A fosfatidiletanolamina é o fosfolipídio mais abundante em células eucarióticas.…”