A method for comprehensive analysis of urinary acylglycines by using ultra-performance liquid chromatography quadrupole linear ion trap mass spectrometry |
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Authors: | Avalyn E Lewis-Stanislaus Liang Li |
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Institution: | aDepartment of Chemistry, University of Alberta, Edmonton, Alberta, Canada |
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Abstract: | Acylglycines are an important class of metabolites that have been used in the diagnosis of several inborn errors of metabolism
(IEM). However, current analytical methods detect only a few acylglycines. There is a need to profile these metabolites in
a comprehensive manner for studying their functions and improving their diagnostic values for different IEM and potentially
other diseases. We describe a sensitive method that combines the chromatographic resolving power of ultra-performance liquid
chromatography (UPLC) to separate closely related metabolites including isomers with tandem mass spectrometry (MS/MS). Acylglycines
were extracted from urine using an anion exchange solid-phase extraction (SPE) cartridge. After UPLC separation, the acylglycines
were detected on a hybrid triple quadrupole linear ion trap mass spectrometer. A set of standards were used for the development
of an optimal MS acquisition method. Several acquisition modes using information derived from collisioninduced dissociation
breakdown curves were used to detect acylglycines. Using this method, 18 acylglycines were detected in the urine of healthy
individuals and confirmed using standards, while 47 additional acylglycines were detected and tentatively identified, based
on their retention and fragmentation pattern. Among the 65 acylglycines detected, only 18 of them have been previously reported
in biofluids of healthy individuals. These results will be deposited in a public human metabolome database. This example illustrates
that by developing a method tailored to the analysis of a class of metabolites sharing similar structural moieties, we can
potentially identify many more new metabolites, thereby expanding the overall metabolome coverage. |
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