Surface-assisted Laser Desorption/ionization Mass Spectrometry Research Articles

Steroid hormones analysis with surface-assisted laser desorption/ionization mass spectrometry using catechin-modified titanium dioxide nanoparticles

This paper describes the application of catechin-modified titanium dioxide nanoparticles (TiO2 NPs) as matrices to analyze four steroid hormones by surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). The catechin-modified TiO2 NPs have high absorbance at 337nm and are effective SALDI matrices when using a nitrogen laser. Four test steroid hormones—cortisone, hydrocortisone, progesterone, and testosterone—were directly analyzed by SALDI-MS. The limits of detection at a signal-to-noise ratio of 3 for cortisone, hydrocortisone, progesterone, and testosterone were 1.62, 0.70, 0.66, and 0.23μM, respectively. This approach provides good quantitative linearity for the four analytes (R2>0.986) with good reproducibility (the shot-to-shot and batch-to-batch variations for the four analytes were less than 10% and 15%, respectively). We validated the practicality of this approach—considering its advantages in sensitivity, repeatability, rapidity, and simplicity—through the analysis of testosterone in a urine sample.

3‐Mercaptopropionic acid modified ZnSe quantum dots as the matrix for direct surface‐assisted laser desorption/ionization mass spectrometric analysis of peptides/proteins from sodium salt solution

This study describes a strategy of using zinc selenium quantum dots (ZnSe QDs) modified with 3-mercaptopropionic acid (3-MPA) as the matrix for direct analysis of peptides and proteins from sodium salt solution in surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). The enhancement of detection sensitivity for these biomolecules was due to the adsorption of positively charged peptides or proteins onto the surfaces of negatively charged ZnSe-3MPA QDs via electrostatic interactions resulting in an increase in ionization efficiency for sodium adduct ions ([M+Na](+)). The applicability of the current approach was demonstrated for a variety of peptides, including leucine-enkephalin, methione-enkephalin, HW6, substance P and angiotensin II, and proteins (cytochrome c, myoglobin and lysozyme). Signal intensities of these peptides or proteins can be enhanced by 25-95 times compared with those obtained by LDI-MS in the absence of ZnSe-3MPA QDs. Applying ZnSe-3MPA QDs to serve as the matrix in SALDI-MS is a simple and effective approach for direct analysis of peptide and protein molecules from sodium salt solution without any pretreatment as the peptides and proteins can be successfully detected as sodium adduct ions ([M+Na](+)).

ChemInform Abstract: Nanoparticle‐Based Mass Spectrometry for the Analysis of Biomolecules

AbstractReview: 60 refs.

Nanoparticle-based mass spectrometry for the analysis of biomolecules

Nanoparticles (NPs) are useful as matrixes for the analyses of several types of biomolecules (including aminothiols, peptides, and proteins) and for mass spectrometric imaging through surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS), mainly because of their large surface area, strong absorption in the ultraviolet-near-infrared region, and ready functionalization. Metallic NPs, metal oxide NPs, and semiconductor quantum dots, unmodified or functionalized with recognition ligands, have a strong affinity toward analytes; therefore, they allow the enrichment of biomolecules, leading to improved sensitivity with minimal matrix interference in their mass spectra. SALDI-MS using NPs overcomes the two major problems commonly encountered in matrix-assisted laser desorption/ionization mass spectrometry: the presence of "sweet spots" and the high background signals in the low-mass region. In this tutorial review, we discuss the roles played by the nature, size, and concentration of the NPs, the buffer composition, and the laser energy in determining the sensitivity and mass ranges for the analytes. We describe internal standard SALDI-MS methods that allow the concentrations of analytes to be determined with low variation (relative standard deviations: <10%) and we highlight how the simplicity, sensitivity, and reproducibility of SALDI-MS approaches using various NPs allow the analyses of proteins and small analytes and the imaging of cells.

Nanocomposites as novel surfaces for laser desorption ionizationmass spectrometry.

The possibility to utilize nanocomposite films as easy-to-handle surfaces for surface assisted laser desorption ionization-mass spectrometry (SALDI-MS) of small molecules, such as pharmaceutical compounds, was evaluated. The signal-to-noise values of acebutolol, propranolol and carbamazepine obtained on the nanocomposite surfaces were higher than the values obtained on plain PLA surface showing that the nanoparticles participate in the ionization/desorption process even when they are immobilized in the polymer matrix. The advantages of nanocomposite films compared to the free nanoparticles used in earlier studies are the ease of handling and reduction of instrument contamination since the particles are immobilized into the polymer matrix. Eight inorganic nanoparticles, titanium dioxide, silicon dioxide, magnesium oxide, hydroxyapatite, montmorillonite nanoclay, halloysite nanoclay, silicon nitride and graphitized carbon black at different concentrations were solution casted to films with polylactide (PLA). There were large differences in signal intensities depending on the type of drug, type of nanoparticle and the concentration of nanoparticles. Polylactide with 10% titanium oxide or 10% silicon nitride functioned best as SALDI-MS surfaces. The limit of detection (LOD) for the study was ranging from 1.7 ppm up to 56.3 ppm and the signal to noise relative standard deviations for the surface containing 10% silicon nitride was approximately 20-30%. Scanning electron microscopy demonstrated in most cases a good distribution of the nanoparticles in the polymer matrix and contact angle measurements showed increasing hydrophobicity when the nanoparticle concentration was increased, which could influence the desorption and ionization. Overall, the results show that nanocomposite films have potential as surfaces for SALDI-MS analysis of small molecules.

Platinum Nanoflowers on Scratched Silicon by Galvanic Displacement for an Effective SALDI Substrate

We report a new and facile method for synthesizing 3D platinum nanoflowers (Pt Nfs) on a scratched silicon substrate by electroless galvanic displacement and discuss the applications of the Pt Nfs in surface-assisted laser desorption/ionization-mass spectrometry (SALDI-MS). Surface scratching of n-type silicon is essential to induce Pt Nf growth on a silicon substrate (to obtain a Pt Nf silicon hybrid plate) by the galvanic displacement reaction. The Pt Nf silicon hybrid plate showed excellent SALDI activity in terms of the efficient generation of protonated molecular ions in the absence of a citrate buffer. We propose that the acidity of the Si-OH moieties on silicon increases because of the electron-withdrawing nature of the Pt Nfs; hence, proton transfer from the Si-OH groups to the analyte molecules is enhanced, and finally, thermal desorption of the analyte ions from the surface occurs. Signal enhancement was observed for protonated molecular ions produced from a titania nanotube array (TNA) substrate on which Pt nanoparticles had been photochemically deposited. Moreover, surface modification of the Pt Nf silicon hybrid plate by perfluorodecyltrichlorosilane (FDTS) (to obtain an FDTS-Pt Nf silicon hybrid plate) was found to facilitate soft SALDI of labile compounds. More interestingly, the FDTS-Pt Nf silicon hybrid plate acts 1) as a high-affinity substrate for phosphopeptides and 2) as a SALDI substrate. The feasibility of using the FDTS-Pt Nf silicon hybrid plate for SALDI-MS has been demonstrated by using a β-casein digest and various analytes, including small molecules, peptides, phosphopeptides, phospholipids, carbohydrates, and synthetic polymers. The hybridization of Pt Nfs with a scratched silicon substrate has been found to be important for achieving excellent SALDI activity.

Detection of Proteins and Protein−Ligand Complexes Using HgTe Nanostructure Matrixes in Surface-Assisted Laser Desorption/Ionization Mass Spectrometry

We have analyzed peptides, proteins, and protein-drug complexes through surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using HgTe nanostructures as matrixes. We investigated the effects of several parameters, including the concentration of the HgTe nanostructures, the pH of the buffer, and the concentration of salt, on the performance of this system. When HgTe nanostructures are used as matrixes, [M + H](+) ions were the dominant signals. Relative to other commonly used nanomaterials, HgTe nanostructures provided lower background signals from metal clusters, fewer fragment ions, less interference from alkali-adducted analyte ions, and a higher mass range (up to 150,000 Da). The present approach provides limits of detection for angiotensin I and bovine serum albumin of 200 pM and 14 nM, respectively, with great reproducibility (RSD: <25%). We validated the applicability of this method through the detections of (i) the recombinant proteins that were transformed in E. coli, (ii) the specific complex between bovine serum albumin and l-tryptophan, and (iii) a carbonic anhydrase-acetazolamide complex. Our results suggest that this novel and simple SALDI-MS approach using HgTe nanostructures as matrixes might open several new ways for proteomics and the analysis of drug-protein complexes.

Nanomaterial-Based Surface-Assisted Laser Desorption/Ionization Mass Spectrometry of Peptides and Proteins

We have investigated six nanomaterials for their applicability as surfaces for the analyses of peptides and proteins using surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). Gold nanoparticles (NPs) were useful nanomaterials for small analytes (e.g., glutathione); Pt nanosponges and Fe(3)O(4) NPs were efficient nanomaterials for proteins, with an upper detectable mass limit of ca. 25 kDa. Nanomaterials have several advantages over organic matrices, including lower limits of detection for small analytes and lower batch-to-batch variations (fewer problems associated with "sweet spots"), when used in laser desorption/ionization mass spectrometry.

Exploring the interactions between gold nanoparticles and analytes through surface‐assisted laser desorption/ionization mass spectrometry

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is applied to provide strong evidence for the chemical reactions of functionalized gold nanoparticles (Au NPs) with analytes--Hg(2+) ions induced MPA-Au NPs aggregation in the presence of 2,6-pyridinedicarboxylic acid (PDCA) and H(2)O(2) induced fluorescence quenching of 11-MUA-Au NDs. PDCA-Hg(2+)-MPA coordination is responsible for Au NPs aggregation, while the formation of 11-MUA disulfide compounds that release into the bulk solution is responsible for H(2)O(2)-induced fluorescence quenching. In addition to providing information about the chemical structures, SALDI-MS is also selective and sensitive for the detection of Hg(2+) ions and H(2)O(2). The limits of detection (LODs) for Hg(2+) ions and H(2)O(2) by SALDI-MS were 300 nM and 250 microM, respectively. The spot-to-spot variations in the two studies were both less than 18% (50 sample spots). Our results reveal that SALDI-MS can be used to study analyte-induced changes in the surface properties of nanoparticles.

Effectiveness of platinum particle deposition on silicon surfaces for surface-assisted laser desorption/ionization mass spectrometry of peptides

One of the problems associated with surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using metal nanoparticles is that peptide analytes are characterized by alkali metal ion adducts even though in the presence of an excess proton source of trifluoroacetic acid in the mass spectra, and their alkali metal ion adducts are often observed in the low peak intensity. In this study, to improve the detection of the proton adduct form of the peptides, the effectiveness of platinum (Pt) particle deposition on a silicon plate for SALDI-MS was examined using the following different Pt-deposited silicon substrates: (1) Pt galvanic deposition (PGDS), (2) Pt particles deposited by suspension deposition on silicon, (3) evaporated Pt on oxidized silicon, and (4) evaporated Pt on bare silicon. Among these substrates, the PGDS plate exhibited the dominant proton adduct form of peptides in the SALDI mass spectra. In addition to platinum deposition, we attempted to employ palladium galvanic deposition as a SALDI-MS substrate, but it was found to lead mainly to the Na+ and K+ forms of the molecular ion. We propose that UV laser radiation-mediated surface-localized positive charges generated by effective charge separation (electron and hole) in the PGDS are important for the production of the proton adduct forms of peptides. Gly-Gly-Tyr-Arg, caffeine, raffinose, and β-cyclodextrin were used as examples to demonstrate the feasibility of PGDS as a matrix-free method for LDI-MS. It was possible to detect these small molecules with SALDI-MS using PGDS; however, the detection sensitivity of these small molecules remains an unresolved issue.

Accurate quantitation of glutathione in cell lysates through surface-assisted laser desorption/ionization mass spectrometry using gold nanoparticles

We developed a method for the determination of three aminothiols—cysteine, glutathione (GSH), and homocysteine—using surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). The analytes were first captured using the unmodified 14-nm gold nanoparticles; N-2-mercaptopropionylglycine–modified gold nanoparticles serving as internal standard were sequentially added, and then the sample was analyzed using SALDI-MS. This approach provided good quantitative linearity of the three analytes ( R 2 = ∼0.99), with good reproducibility (relative standard deviations: <10%), in the analyses of GSH in the lysates of human red blood cells and MCF-7 cancer breast cells in the presence and absence of the anti-inflammatory drug sulfasalazine. The internal-standard SALDI-MS approach provides simplicity, accuracy, and precision to the determination of GSH in cells under drug invasion, to open an avenue for SALDI-MS to be used for the precise quantitative determination of a variety of analytes. From the Clinical Editor This paper reports the development of a surface assisted laser desorption/ionization mass spectrometry method to precisely determine aminothiols-cysteine (Cys), glutathione (GSH), and homocysteine (HCys)

Quantification of Captopril in Urine Through Surface-Assisted Laser Desorption/Ionization Mass Spectrometry Using 4-Mercaptobenzoic Acid-Capped Gold Nanoparticles as an Internal Standard

We have developed a new internal standard method for the determination of the concentration of captopril (CAP) through surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using gold nanoparticles (Au NPs). This approach provided linearity for CAP over the concentration range 2.5-25 microM (R(2) = 0.987), with a limit of detection (signal-to-noise ratio = 3) of 1.0 microM. The spot-to-spot variations in the concentration of CAP through SALDI-MS analyses performed in the absence and presence of the internal standard were 26% and 9%, respectively (15 measurements). This approach provides simplicity, accuracy, precision, and great reproducibility to the determination of the levels of CAP in human urine samples.

Quantitative Analysis of an Antioxidant Additive in Insoluble Plastics by Surface-Assisted Laser Desorption/Ionization Mass Spectrometry (SALDI-MS) Using TiO2 Nanoparticles

The applicability of matrix-free surface-assisted laser desorption/ionization mass spectrometry using porous silicon (DIOS-MS) and TiO2 nanoparticles (TiO2-SALDI-MS) was examined by analyzing an antioxidant added to insoluble polypropylene (PP) materials. The optimized solvent extraction, which involved freezing and crushing the PP, results in a good recovery, 89±4 and 97±8% for 0.5 wt% of the added antioxidant, tetrakis[methylene3-(3,5-di-t-butyl-4-hydroxyphenyl)propionate]methane (commercial name; Irganox1010) in laboratory-produced and commercial PP composites, respectively. The positive ion measurements in DIOS and TiO2-SALDI-MS yielded sodium adduct ions without any interference from other signals. Using an internal standard with a similar structure, a good calibration curve linearity was established in the Irganox1010 concentration range of 0.01-2.0 mg/mL in PP, which was sufficient for monitoring the plastic additives. However, TiO2-SALDI-MS was superior to DIOS-MS in terms of mass resolution and easiness of sample preparation. A solvent extraction method followed by TiO2-SALDI-MS appears to be a anticipated method for the analysis of additives in insoluble plastics.

Screening and Quantification of Pesticides in Water Using a Dual-Function Graphitized Carbon Black Disk

A simple platform for combining solid phase extraction (SPE) and surface-assisted laser desorption ionization mass spectrometry (SALDI-MS) of extracted analytes, using disks prepared by embedding graphitized carbon black (GCB-4) particles in a network of polytetrafluoroethylene (PTFE), is presented. The system provides a convenient approach for rapid SALDI-MS screening of substances in aqueous samples, which can be followed by robust quantitative and/or structural analyses by liquid chromatography (LC)/MS/MS of positive samples. The extraction discs are easily transferred between gaskets where the sample extraction and desorption of selected samples is performed and the mass spectrometer. The SPE and SALDI properties of the new GCB-4 disc have been characterized for 15 pesticides with varying chemical properties, and the screening strategy has been applied to the analysis of pesticides in agricultural drainage water. Atrazine and atrazine-desethyl-2-hydroxy were detected in the sampled water by SALDI-MS screening and subsequently confirmed and quantified using LC/MS/MS.

Functionalized pyrolytic highly oriented graphite polymer film for surface‐assisted laser desorption/ionization mass spectrometry in environmental analysis

The pyrolytic highly oriented graphite polymer film (PGS) was first employed to analyze low-mass analytes in environmental analysis by surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). PGS is a synthetic uniform and highly oriented graphite polymer film with high thermal anisotropic conductivity. We have found that negative ion mode SALDI-MS using oxidized PGS (PGS-SALDI-MS) can be used to detect [M-H]- ions from perfluorooctanoic acid (PFOA) and other perfluoroalkylcarboxylic acids when the PGS surface is modified with the cationic polymer polyethyleneimine (PEI). The signal intensity of PFOA when employing the PEI modification showed a ten-fold increase over that obtained from desorption/ionization on porous silicon (DIOS). PFOA was quantified using PGS-SALDI-MS and the calibration curve showed a wide linear dynamic range of response (20-1000 ppb). The combination of atmospheric pressure ionization and PGS (AP-PGS-SALDI) showed greater signal intensity than vacuum PGS-SALDI for deprotonated PFOA. Several other environmentally important chemicals, including perfluoroalkylsulfonic acid, pentachlorophenol, bisphenol A, 4-hydroxy-2-chlorobiphenyl, and benzo[a]pyrene, were also successfully used to evaluate PGS-SALDI-MS. In addition, we found that nonafluoro-1-butanesulfonic acid was able to produce protonated peptides in positive ion PGS-SALDI-MS, but that perfluoropentanoic acid and trifluoroacetic acid were not. It is suggested that perfluoroalkylsulfonic acids are better protonating agents than perfluoroalkylcarboxylic acids in SALDI-MS.

Detection of aminothiols through surface‐assisted laser desorption/ionization mass spectrometry using mixed gold nanoparticles

We have employed mixtures of two differently sized (average diameters: 3.5 and 14 nm) gold nanoparticles (Au NPs) as selective probes and matrices for the determination of aminothiols using surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). When using 38 and 150 pM solutions of the 3.5- and 14-nm Au NPs, respectively, as the probe and matrix, SALDI-MS provided limits of detection (signal-to-noise ratio = 3) of 2, 20, and 44 nM for 1.0 mL solutions of glutathione (GSH), cysteine (Cys), and homocysteine, respectively. The signal intensities of these analytes varied by less than 20% for SALDI-MS analyses recorded over 50 sample spots; in contrast, they varied by as much as 60% when using a conventional matrix (2,5-dihydroxybenzoic acid). We validated the practicality of this approach - with its advantages of sensitivity, reproducibility, rapidity, and simplicity - through the analysis of GSH in MCF-7 cell lysates and Cys in plasma.

Effect of urea surface modification and photocatalytic cleaning on surface‐assisted laser desorption ionization mass spectrometry with amorphous TiO2 nanoparticles

We have investigated the effect of urea surface modification and the photocatalytic cleaning on surface-assisted laser desorption ionization mass spectrometry (SALDI-MS) with amorphous TiO2 nanoparticles for the reduction of the background noise and the improvement of the sensitivity. In the use of nanoparticles of high surface area, chemical background signals arising from ambient environments and organic contaminants can frequently be serious problems below 500 Da, possibly reducing the advantages of the matrix-free approach. In this study, removal of contaminants and enhanced SALDI efficiency were easily achieved with UV irradiation via the photocatalyst effect of TiO2 before SALDI-MS measurements. The surface cleaning achieved by the UV photocatalytic procedure reduced the background noise and increased the peak intensities of peptides. In addition, we found that urea surface modification of TiO2 nanoparticles increased the performance of the TiO2-SALDI-MS. (1) The urea-surface modification of TiO2 made it possible to produce proton-adduct forms without citrate buffer, resulting in low background noises below 500 Da, in contrast to the essential use of a citrate buffer in the bare TiO2-SALDI-MS. (2) The detection sensitivity of angiotensin I increased to 0.3 fmol with the urea-surface modification, as compared to the use of bare TiO2 nanoparticles (6 fmol). The urea-TiO2 could ionize proteins of more than 20,000 Da such as trypsinogen (600 fmol). (3) The urea modification of TiO2 had the advantage of selective detection of phosphopeptides without sample clean up, or prefractionation in tryptic digest products of bovine hemoglobin.

Ion Desorption Efficiency and Internal Energy Transfer in Carbon-Based Surface-Assisted Laser Desorption/Ionization Mass Spectrometry: Desorption Mechanism(s) and the Design of SALDI Substrates

Ion desorption efficiency and internal energy transfer were probed and correlated in carbon-based surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using benzylpyridinium (BP) salt as the thermometer chemical. In a SALDI-MS experiment with a N(2) laser (at 337 nm) used as the excitation light source and with multiwalled carbon nanotubes (CNT), buckminsterfullerene (C(60)), nanoporous graphitic carbon (PGC), non-porous graphite particles (G), highly oriented pyrolytic graphite (HOPG), or nanodiamonds (ND) as the SALDI substrate, both the desorption efficiency in terms of ion intensity of BP and the extent of internal energy transfer to the ions are dependent on the type and size of the carbon substrates. The desorption efficiency (CNT approximately C(60) > PGC > G > HOPG > ND) in general exhibits an opposite trend to the extent of internal energy transfer (CNT < C(60) approximately PGC < G approximately HOPG < ND), suggesting that increasing the extent of internal energy transfer in the SALDI process may not enhance the ion desorption efficiency. This phenomenon cannot be explained by a thermal desorption mechanism, and a non-thermal desorption mechanism is proposed to be involved in the SALDI process. The morphological change of the substrates after the laser irradiation and the high initial velocities of BP ions (1100-1400 ms(-1)) desorbed from the various carbon substrates suggest that phase transition/destruction of substrates is involved in the desorption process. Weaker bonding/interaction and/or a lower melting point of the carbon substrates favor the phase transition/destruction of the SALDI substrates upon laser irradiation, consequently affecting the ion desorption efficiency.

Detailed Investigation on the Possibility of Nanoparticles of Various Metal Elements for Surface-Assisted Laser Desorption/Ionization Mass Spectrometry

In this paper, we describe systematic detailed considerations of the feasibility of using various metal nanoparticles for organic-matrix-free surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). In order to avoid the influence of organic molecules on the nanoparticles, stabilizer-free bare nanoparticles of Ag, Au, Cu and Pt were prepared by laser ablation. Although all metal nanoparticles absorbed N(2) laser light (337 nm) energy, the performance of desorption/ionization of a representative peptide, angiotensin I, strongly depended on the metal element. Citrate buffer was used as a proton source; it reduced the amount of alkali cation adducts present. Then, protonated molecules of analytes predominated in the mass spectra when Au and Pt nanoparticles were used. Pt nanoparticles showed the highest performance in SALDI-MS, owing to their smaller heat conductivity and higher melting temperature. The selective desorption of a cationic surfactant with longer alkyl chains and a peptide with methionine was also observed.

Surface cleaning of germanium nanodot ionization substrate for surface‐assisted laser desorption/ionization mass spectrometry

In surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS), a chemical background signal, arising from organic contaminants such as plasticizers, is frequently observed mainly under m/z ca. 600, which impairs the advantages of the matrix-free approach. Silver salts, which are used for the cationization of aromatic compounds, are also difficult to remove completely after the measurements. In this study, surface cleaning techniques used in semiconductor processing were used to clean our developed silicon-based SALDI substrate on which self-assembled germanium nanodots (GeNDs) had been deposited (termed a GeND chip). An immersion cleaning method using acetone with sonication, and a sulfuric-peroxide mixture (SPM) cleaning method using a mixture of H(2)SO(4)/H(2)O(2)/deionized water, were examined for their effectiveness in removing organic compounds and residual silver salts. Removal of both types of contaminants was successfully performed by SPM cleaning. The limit of detection for glutathione was improved from ca. 5 pmol without cleaning to ca. 50 fmol after the SPM cleaning. Since GeND chips can tolerate acidic cleaning and sonication due to their chemical inertness and rigid nanodot structures, they appear to be an ideal reusable SALDI substrate.