We report that modifying the surface of gold nanoparticles (Au NPs) with 2-mercaptopyridine-3-carboxylic acid (MPyCA) enhances surface-assisted laser desorption/ionization (SALDI) performance in the analysis of small molecules. The MPyCA ligand has a strong UV absorbance at the wavelengths of the typical MALDI laser at 337 nm, resulting in efficient thermal/energy transfer from the Au NPs to analytes during pulse-laser irradiation. In addition, the MPyCA ligand contains carboxylic acid and pyridine groups, providing affinity to various analytes through acid-base interactions.

Irganox1010, glucose and meropenem were utilized as model analytes to evaluate SALDI performance because these molecules are generally ionized with difficulty by conventional MALDI-MS. Our results demonstrate that the MPyCA-Au NP based SALDI-MS could detect Irganox1010, glucose and meropenem with stronger ion peaks for these molecules compared to MALDI-MS using CHCA. The limit of detection (LOD) for meropenem was much lower in the case of SALDI (LOD=1 ng/mL) compared to MALDI (LOD=10 μg/mL).

We have developed a rapid and sensitive analytical method for α-tocopherol and its oxidative products by combining online hyphenation of supercritical fluid extraction-supercritical fluid chromatography (SFC) with proton transfer reaction (PTR) ionization mass spectrometry (MS). α-Tocopherol is a well-known antioxidant that plays a vital role in the antioxidant defense system in plant cells. However, studies on the cellular mechanisms of α-tocopherol have been limited owing to the lack of a rapid analytical method, which limits the comparison of plant cells incubated in various conditions. Additionally, complex sample preparation and long chromatography separation times are required. Moreover, the majority of the involved molecules are a combination of isomers, which must be separated before applying tandem MS. α-Tocopherol produces the α-tocopheroxyl radical in the first step of its antioxidant function; another ion with the same mass may also be generated from the source. SFC separation effectively distinguished the observed ions from their oxidative products in the sample and those produced during the ionization reaction process. This method enabled the measurement of α-tocopherol and its oxidative products such as α-tocopheroxyl radical and α-tocopheryl quinone in approximately 3 min per sample, including the time required for sample preparation.

The insertion of ion mobility spectrometry (IMS) between LC and MS can improve peptide identification in both proteomics and phosphoproteomics by providing structural information that is complementary to LC and MS, because IMS separates ions on the basis of differences in their shapes and charge states. However, it is necessary to know how phosphate groups affect the peptide collision cross sections (CCS) in order to accurately predict phosphopeptide CCS values and to maximize the usefulness of IMS. In this work, we systematically characterized the CCS values of 4,433 pairs of mono-phosphopeptide and corresponding unphosphorylated peptide ions using trapped ion mobility spectrometry (TIMS). Nearly one-third of the mono-phosphopeptide ions evaluated here showed smaller CCS values than their unphosphorylated counterparts, even though phosphorylation results in a mass increase of 80 Da. Significant changes of CCS upon phosphorylation occurred mainly in structurally extended peptides with large numbers of basic groups, possibly reflecting intramolecular interactions between phosphate and basic groups.