Amino Acid Profiling by Reactive Desorption Electrospray Ionization Tandem Mass Spectrometry (DESI-MS/MS)

Desorption electrospray ionization mass spectrometry (DESI MS) has gained significant recognition over the past few years because of its simplicity and rapid throughput capabilities, allowing for the direct analysis of samples with a wide variety of sizes, shapes, and chemistries. Addition of various reagents into the DESI spray solvent enables atmospheric pressure ion molecule reactions between these reagents in the charged micro droplets and analyte molecules on the sample surface affording improved selectivity and sensitivity in some cases. Presented is a rapid screening method for amino acids (aas) based on reactive DESI. Amino acids have been shown to play key roles in the regulation of cellular processes. They are also particularly vital in the determination of metabolic disorders such as phenylketonurea, homocystinuria, and tyrosinemia. The most specific and reliable methods for diagnosing these disorders are based on the determination of aas in body fluids using methodologies such as gas/liquid chromatography, tandem MS, and various combinations thereof. However, these methods are usually time-consuming, increasing the time physicians wait before administering treatment or regulating the diet of diseased infants. The reactive DESI approach presented here is based on the formation of stable noncovalent complexes between alpha-cyclodextrin (dissolved in the spray solvent) and amino acids present in the sample affording a selective method for their detection. However, the selectivity and sensitivity of screening for aas was improved by performing the MS analysis in the multiple reaction monitoring mode when using a quadrupole ion trap or by the precursor ion scan when using a triple quadrupole MS instrument, affording an average twenty-five times sensitivity improvement compared to analysis in 3 full scan mode. The observation of similar complexes with various carboxylic acids including formic acid and acetic acid, and evidence from tandem MS experiments indicate that amino acid:alpha-cyclodextrin complexation reactions occur by hydrogen bonding interaction with carboxyl group of the aas. The specificity and sensitivity provided by this approach seems very promising for applications in the rapid screening of aas directly from body fluids including urine and plasma for amino acid disorders in a clinical setting.