Surface-assisted Laser Desorption/ionization Mass Spectrometry Research Articles

Gold nanoparticles modified porous silicon chip for SALDI-MS determination of glutathione in cells

The gold nanoparticles (Au NPs) modified porous silicon chip based surface assisted laser desorption/ionization mass spectrometry (SALDI-MS) was developed to capture and analyze glutathione (GSH) in cells. With silver-assisted chemical etching, Ag nanoparticles (Ag NPs) were generated and deposited on the silicon surface and the nanopores were etched on silicon substrate. Then Au NPs were in-situ synthesized on the ridges of silicon nanopores. This Ag-Au NPs modified porous silicon surface could specially capture and enrich thiol compounds through Au-S binding, and it could also function as matrix to assist ionization for SALDI-MS. The silicon chip was array patterned for high throughput SALDI-MS detection. GSH and cysteine could be distinguished without the interference from matrix signals. This approach was successfully applied to preconcentration and detection of GSH in Caco-2 cells. The GSH alterations in cells under drug stimulation were investigated. This invented silicon chip showed great potential for more efficient analysis of small thiol biomarkers in complex biological samples.

Fabrication of Nanostructured Mesoporous Germanium for Application in Laser Desorption Ionization Mass Spectrometry.

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is a high-throughput analytical technique ideally suited for small-molecule detection from different bodily fluids (e.g., saliva, urine, and blood plasma). Many SALDI-MS substrates require complex fabrication processes and further surface modifications. Furthermore, some substrates show instability upon exposure to ambient conditions and need to be kept under special inert conditions. We have successfully optimized mesoporous germanium (meso-pGe) using bipolar electrochemical etching and efficiently applied meso-pGe as a SALDI-MS substrate for the detection of illicit drugs such as in the context of workplace, roadside, and antiaddictive drug compliance. Argon plasma treatment improved the meso-pGe efficiency as a SALDI-MS substrate and eliminated the need for surface functionalization. The resulting substrate showed a precise surface geometry tuning by altering the etching parameters, and an outstanding performance for illicit drug detection with a limit of detection in Milli-Q water of 1.7 ng/mL and in spiked saliva as low as 5.3 ng/mL for cocaine. The meso-pGe substrate had a demonstrated stability over 56 days stored in ambient conditions. This proof-of-principle study demonstrates that meso-pGe can be reproducibly fabricated and applied as an analytical SALDI-MS substrate which opens the door for further analytical and forensic high-throughput applications.

Gold nanoparticles bridging infra-red spectroscopy and laser desorption/ionization mass spectrometry for direct analysis of over-the-counter drug and botanical medicines

With a coating of gold nanoparticles (AuNPs), over-the-counter (OTC) drugs and Chinese herbal medicine granules in KBr pellets could be analyzed by Fourier Transform Infra-red (FT-IR) spectroscopy and Surface-assisted Laser Desorption/Ionization mass spectrometry (SALDI-MS). FT-IR spectroscopy allows fast detection of major active ingredient (e.g., acetaminophen) in OTC drugs in KBr pellets. Upon coating a thin layer of AuNPs on the KBr pellet, minor active ingredients (e.g., noscapine and loratadine) in OTC drugs, which were not revealed by FT-IR, could be detected unambiguously using AuNPs-assisted LDI-MS. Moreover, phytochemical markers of Coptidis Rhizoma (i.e. berberine, palmatine and coptisine) could be quantified in the concentrated Chinese medicine (CCM) granules by the SALDI-MS using standard addition method. The quantitative results matched with those determined by high-performance liquid chromatography with ultraviolet detection. Being strongly absorbing in UV yet transparent to IR, AuNPs successfully bridged FT-IR and SALDI-MS for direct analysis of active ingredients in the same solid sample. FT-IR allowed the fast analysis of major active ingredient in drugs, while SALDI-MS allowed the detection of minor active ingredient in the presence of excipient, and also quantitation of phytochemicals in herbal granules.

Graphene flakes enhance the detection of TiO2-enriched catechins by SALDI-MS after microwave-assisted enrichment

In this study, we describe a novel method for the rapid detection of catechins in tea leaves using surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). The method involves incubating catechins in a mixture of titanium dioxide (TiO2) nanoparticles and graphene flakes (GF) and then subjecting the mixture to microwave irradiation to facilitate the enrichment of analytes. In this approach, TiO2 nanoparticles serve as probes to enrich catechins while GF serve to increase the desorption/ionization efficiency. The use of the TiO2–GF mixture as matrix provides a number of advantages over TiO2 nanoparticles alone, including higher desorption/ionization efficiency, enhanced analyte enrichment and reduced run time (less 10min). We demonstrate that the addition of GF to nanoparticles of TiO2 improves the detection of catechins by SALDI-MS. At a signal-to-noise ratio of 3, the limits of detection were 0.04pmole for (+)-catechin (C), 0.17pmole for (–)-epigallocatechin (EGC), 0.35pmole for (–)-epicatechin gallate (ECG), and 0.50pmole for (–)-epigallocatechin gallate (EGCG). This method was further applied to determine the catechin content in several tea samples and the results showed good shot-to-shot and sample-to-sample reproducibility (less than 10% and 13%, respectively) as well as good quantitative linearity for the four analytes (R2>0.995). The precision calculated as percent relative standard derivation (%RSD), ranged from 3.6% to 12.4%. Overall, we show that this method is a sensitive, simple and high-throughput technique for the detection of catechins in tea.

Nanoporous Carbons Derived from Metal‐Organic Frameworks as Novel Matrices for Surface‐Assisted Laser Desorption/Ionization Mass Spectrometry

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) represents a powerful tool for the analysis of biomolecules, synthetic polymers, and even small organic compounds; its performances largely depend on the type of matrix materials utilized. Here, for the first time the employment of nanoporous carbons derived from metal-organic frameworks (MOFs) as novel matrices for SALDI-MS is demonstrated. The nanoporous carbons derived from MOFs not only circumvent the shortcomings of existing matrix materials but also demonstrate much higher efficiency of laser desorption/ionization for various compounds than any other nanoporous carbons reported so far. A new perspective for the development of matrix materials for SALDI-MS application is therefore provided.

Identification of Microalgae by Laser Desorption/Ionization Mass Spectrometry Coupled with Multiple Nanomatrices

In this study, we demonstrate a simple method to identify microalgae by surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using three different substrates: HgSe, HgTe, and HgTeSe nanostructures. The fragmentation/ionization processes of complex molecules in algae varied according to the heat absorption and transfer efficiency of the nanostructured matrices (NMs). Therefore, the mass spectra obtained for microalgae showed different patterns of m/z values for different NMs. The spectra contained both significant and nonsignificant peaks. Constructing a Venn diagram with the significant peaks obtained for algae when using HgSe, HgTe, and HgTeSe NMs in m/z ratio range 100-1000, a unique relationship among the three sets of values was obtained. This unique relationship of sets is different for each species of microalgae. Therefore, by observing the particular relationship of sets, we successfully identified different algae such as Isochrysis galbana, Emiliania huxleyi, Thalassiosira weissflogii, Nannochloris sp., Skeletonema cf. costatum, and Tetraselmis chui. This simple and cost-effective SALDI-MS analysis method coupled with multi-nanomaterials as substrates may be extended to identify other microalgae and microorganisms in real samples. Graphical Abstract Identification of microalgae by surface-assisted laser desorption/ionization mass spectrometry coupled with three different mercury-based nanosubstrates.

A novel type of matrix for surface-assisted laser desorption–ionization mass spectrometric detection of biomolecules using metal-organic frameworks

A 3D metal-organic framework (MOF) nanomaterial as matrix for surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) and tandem mass spectrometry (MS/MS) was developed for the analysis of complex biomolecules. Unlike other nanoparticle matrices, this MOF nanomaterial does not need chemical modification prior to use. An exceptional signal reproducibility as well as very low background interferences in analyzing mono-/di-saccharides, peptides and complex starch digests demonstrate its high potential for biomolecule assays, especially for small molecules.

Impact of analyte ablation and surface acidity of Pd nanoparticles on efficiency of surface-assisted laser desorption/ionization-mass spectrometry

In this study, the impact of analyte ablation and substrate surface acidity on ion yield in surface-assisted laser desorption/ionization-mass spectrometry (SALDI-MS) from Pd nanoparticles was examined. The extent of analyte removal from the Pd material was investigated by X-ray analysis and compared to conventional desorption/ionization on silicon (DIOS) and matrix-assisted laser desorption/ionization (MALDI). Despite the different surface morphologies of the investigated materials and the widely varying ionization yields observed during the experiments, virtually the same amount of analyte was removed by laser irradiation from each of the investigated substrates at a given laser fluence. Our experiments therefore suggested that the extent of analyte ablation from the substrate was less important for SALDI efficiency than the chemical properties of the surface.

Analyses of functional polymer-modified nanoparticles for protein sensing by surface-assisted laser desorption/ionization mass spectrometry coupled with HgTe nanomatrices

In this study, we employed HgTe nanostructure-based matrices (nanomartrices; NMs) for surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) for the analyses of polyethylene glycol (PEG) derivatives as well as thiol-PEG-modified gold nanoparticles (PEG-Au NPs). Relative to common organic matrices, the use of HgTe NMs as the matrix for SALDI-MS resulted in more highly efficient analyses of PEG derivatives, in terms of sensitivity and reproducibility. The symmetric MS profiles of PEG (Mw: ca. 8000Da) obtained through HgTe NMs/SALDI-MS analysis revealed the absence of polymer degradation during this process. Under optimal conditions, the HgTe NMs/SALDI-MS system enabled the detection of PEG sample as low as 100pg and with molecular weights of up to approximately 42,000Da. We also used this approach for the analyses of PEG-Au NPs in which various functional groups (carboxymethyl, amine, biotin) were present at the PEG termini, revealing that the combination of SALDI-MS and HgTe NMs have great potential for use in the characterization of modified polymer–ligands on nanomaterials. We also demonstrated the PEG-Au NPs can be coupled with HgTe NMs/SALDI-MS for characterization of biorecognition events. After avidin, the target protein, had been selectively captured by the biotin-PEG-Au NPs, we found that the desorption/ionization efficiency of biotin-PEG from the Au NP surface was suppressed; accordingly, this novel SALDI-MS approach allows rapid detection of avidin with high specificity and sensitivity. Au NP surfaces functionalized with other functional-PEG ligands might also allow amplification of signals from other biological interactions.

Synthesis of a highly dispersive sinapinic acid@graphene oxide (SA@GO) and its applications as a novel surface assisted laser desorption/ionization mass spectrometry for proteomics and pathogenic bacteria biosensing.

Graphene oxide (GO)-modified sinapinic acid (3,5-dimethoxy-4-hydroxycinnamic acid, SA) (SA@GO) was synthesized and characterized; it was then investigated as a new surface assisted laser desorption/ionization mass spectrometry (SALDI-MS) for proteomics and pathogenic bacteria biosensing. SA@GO could effectively decrease the time necessary for sweet spotting searching, reducing the amount of organic matrix and solvent and enhance the sensitivity. SA@GO shows high performance as a matrix alone without the need to add trifluoroacetic acid (TFA). However, the analysis of the intact bacteria cells shows improvement in the signal intensity (2-5 fold) and offers a low limit of detection. All these analyses could be performed with low concentrations (1-10 fmol) and tiny volumes (0.5-1 μL). This study demonstrated that the exploration of new hybrid materials is pivotal to achieve high performance and high ionization. Because of the plane of GO, it assists protein-protein interactions that make it undergo softer ionization.

Direct detection of illicit drugs from biological fluids by desorption/ionization mass spectrometry with nanoporous silicon microparticles.

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is a high throughput analytical technique capable of detecting low molecular weight analytes, including illicit drugs, and with potential applications in forensic toxicology as well as athlete and workplace testing, particularly for biological fluids (oral fluids, urine and blood). However, successful detection of illicit drugs using SALDI-MS often requires extraction steps to reduce the inherent complexity of biological fluids. Here, we demonstrate an all-in-one extraction and analytical system consisting of hydrophobically functionalized porous silicon microparticles (pSi-MPs) for affinity SALDI-MS of prescription and illicit drugs. This novel approach allows for the analysis of drugs from multiple biological fluids without sample preparation protocols. The effect of pSi-MP size, pore diameter, pore depth and functionalization on analytical performance is investigated. pSi-MPs were optimized for the rapid and high sensitivity detection of methadone, cocaine and 3,4-methylenedioxymethamphetamine (MDMA). This optimized system allowed extraction and detection of methadone from spiked saliva and clinical urine samples. Furthermore, by detecting oxycodone in additional clinical saliva and plasma samples, we were able to demonstrate the versatility of the pSi-MP SALDI-MS technique.

Matrix-enhanced surface-assisted laser desorption/ionization mass spectrometry (ME-SALDI-MS) for mass spectrometry imaging of small molecules.

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS), a parallel technique to matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), utilizes inorganic particles or porous surfaces to aid in the desorption/ionization of low-molecular-weight (MW) analytes. As a matrix-free and "soft" LDI approach, SALDI offers the benefit of reduced background noise in the low MW range, allowing for easier detection of biologically significant small MW species. Despite the inherent advantages of SALDI-MS, it has not reached comparable sensitivity levels to MALDI-MS. In relation to mass spectrometry imaging (MSI), intense efforts have been made in order to improve sensitivity and versatility of SALDI-MSI. We describe herein a detailed protocol that utilizes a hybrid LDI method, matrix-enhanced SALDI-MS (ME-SALDI MS), to detect and image low MW species in an imaging mode.

Comparison of the performance of different silicon-based SALDI substrates for illicit drug detection

Surface-assisted laser desorption ionization mass spectrometry (SALDI-MS) is an emerging technique used for the detection of small molecules (<700Da) such as illicit drugs. In recent times, this technique has been employed for the detection of illicit drugs in various body fluids including saliva. Three common SALDI techniques, desorption ionization on porous silicon (DIOS), nanostructure-initiator mass spectrometry (NIMS) and nanostructured laser desorption ionization (NALDI™) are compared for the detection of four drug classes, amphetamines, benzodiazepines, opiates and tropane alkaloids. We focus in our comparison on structural and chemical characteristics, as well as analytical performance and longevity.

Functional microgels assisted tryptic digestion and quantification of cytochrome c through internal standard mass spectrometry.

Quantitation of cytochrome c (Cyt c) in cell lysates through surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using gold nanoparticles (Au NPs) as the matrix and GR-10 peptide as an internal standard has been demonstrated. To shorten digestion time, temperature sensitive microgels containing trypsin (TR) and Au NPs have been employed. As-prepared functional microgels (TR/Au NPs/MGs) allow digestion of Cyt c within 15s under microwave irradiation. The internal standard SALDI-MS approach provides linearity (R(2) = 0.98) of MS signal ratio (I 1168.6/I 1067.6) of the tryptic digested peptide (m/z 1168.6) to GR-10 peptide (m/z 1067.6) against the concentration of Cyt c ranging from 25 to 200nM, with a limit of detection (at a signal-to-noise ratio of 3) of 10nM. This approach has been validated by the analysis of the lysates of HeLa cells, with an average concentration of 13.7 ± 3.5μM for cytoplasmic Cyt c. Increased concentrations of Cyt c in the HeLa cells treated with etoposide (a commercial drug) or carbon dots (potential drug) have been revealed through this simple, sensitive, and rapid SALDI-MS approach, supporting the drugs induced Cyt c-mediated apoptosis of the cells. This study has shown that this internal standard SALDI-MS approach holds great potential for cell study.

Surface-assisted laser desorption/ionization mass spectrometry using ordered silicon nanopillar arrays.

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is ideally suited for the high-throughput analysis of small molecules in bodily fluids (e.g. saliva, urine, and blood plasma). A key application for this technique is the testing of drug consumption in the context of workplace, roadside, athlete sports and anti-addictive drug compliance. Here, we show that vertically-aligned ordered silicon nanopillar (SiNP) arrays fabricated using nanosphere lithography followed by metal-assisted chemical etching (MACE) are suitable substrates for the SALDI-MS detection of methadone and small peptides. Porosity, length and diameter are fabrication parameters that we have explored here in order to optimize analytical performance. We demonstrate the quantitative analysis of methadone in MilliQ water down to 32 ng mL(-1). Finally, the capability of SiNP arrays to facilitate the detection of methadone in clinical samples is also demonstrated.

Decoration of silicon nanostructures with copper particles for simultaneous selective capture and mass spectrometry detection of His-tagged model peptide.

We present in this work a simple and fast preparation method of a new affinity surface-assisted laser/desorption ionization mass spectrometry (SALDI-MS) substrate based on silicon nanostructures decorated with copper particles. The silicon nanostructures were fabricated by the metal-assisted chemical etching (MACE) method. Then, superhydrophilic areas surrounded by superhydrophobic regions were formed through hydrosilylation reaction of 1-octadecene, followed by local degradation of the octadecyl layer. After that, copper particles were deposited in the hydrophilic areas by using the electroless method. We have demonstrated that these surfaces were able to perform high selective capture of model His-tag peptide even in a complex mixture such as serum solution. Then, the captured peptide was detected by mass spectrometry at a femtomolar level without the need of organic matrix.

Gold-decorated titania nanotube arrays as dual-functional platform for surface-enhanced Raman spectroscopy and surface-assisted laser desorption/ionization mass spectrometry.

In this report, we demonstrate gold-decorated titania nanotube arrays (Au-TNA substrate) as a dual-functional platform for surface-enhanced Raman spectroscopy (SERS) and surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). The Au nanoparticles are grown on the substrate using vapor deposition of Au. The resulting substrates perform better than Au colloids in terms of the reproducibility of the SERS measurements, long-term stability of the fabricated structures, and clean surface of the Au. The nanostructure of the Au-TNA substrate was designed to optimize the SALDI-MS and SERS performance. Excellent reproducibility of the SERS measurements using the Au-TNA substrate was obtained, with a standard error less than 6 %. SALDI activity was also demonstrated for the same Au-TNA substrates. Finally, the Au-TNA substrate was used for combined SERS and SALDI-MS analysis (i) to discriminate the structural isomers of pyridine compounds (para-, meta-, and ortho-pyridinecarboxylic acid) and (ii) to detect polycarbamate, a dithiocarbamate fungicide. These results are difficult to obtain using either approach alone.

Quantification of saccharides in honey samples through surface-assisted laser desorption/ionization mass spectrometry using HgTe nanostructures.

Quantification of monosaccharides and disaccharides in five honey samples through surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using HgTe nanostructures as the matrix and sucralose as an internal standard has been demonstrated. Under optimal conditions (1× HgTe nanostructure, 0.2mM ammonium citrate at pH9.0), the SALDI-MS approach allows detection of fructose and maltose at the concentrations down to 15 and 10μM, respectively. Without conducting tedious sample pretreatment and separation, the SALDI-MS approach allows determination of the contents of monosaccharides and disaccharides in honey samples within 30min, with reproducibility (relative standard deviation <15%). Unlike only sodium adducts of standard saccharides detected, sodium adducts and potassium adducts with differential amounts have been found among various samples, showing different amounts of sodium and potassium ions in the honey samples. The SALDI-MS data reveal that the contents of monosaccharides and disaccharides in various honey samples are dependent on their nectar sources. In addition to the abundant amounts of monosaccharides and disaccharides, oligosaccharides in m/z range of 650 - 2700 are only detected in pomelo honey. Having advantages of simplicity, rapidity, and reproducibility, this SALDI-MS holds great potential for the analysis of honey samples.

Sensitive Detection of Aromatic Hydrophobic Compounds in Water and Perfluorooctane Sulfonate in Human Serum by Surface-Assisted Laser Desorption/Ionization Mass Spectrometry (SALDI-MS) with Amine Functionalized Graphene-Coated Cobalt Nanoparticles.

In this article, we describe the application of surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) with the use of amine functionalized graphene-coated cobalt nanoparticles (CoC-NH2 nanoparticles) to analyse aromatic hydrophobic compounds that are known environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs) and pentachlorophenol (PCP). Our results demonstrated that SALDI-MS can detect PCP, anthracene, and pyrene in water. In particular, the CoC-NH2 nanoparticles proved to be an efficient means of capturing PCP in water because of the high adsorption capacity of the nanoparticles for PCP, which resulted in a detectability of 100 ppt. Furthermore, the CoC-NH2 nanoparticles also functioned as an adsorbent for solid-phase extraction of perfluorooctane sulfonate (PFOS) from human serum, displaying good performance with a detectability of 10 ppb by SALDI-MS.

Laser-Based Mass Spectrometric Determination of Aggregation Numbers for Petroleum- and Coal-Derived Asphaltenes

Petroleum- and coal-derived asphaltenes have been studied with three laser-based mass spectrometric techniques: laser desorption ionization–mass spectrometry (LDI–MS), in which a single laser desorbs and ionizes solid analytes; surface-assisted laser desorption ionization–mass spectrometry (SALDI–MS), in which a single laser desorbs and ionizes solid analytes from an activated surface; and laser desorption laser ionization mass spectrometry (L2MS), in which desorption and ionization are separated spatially and temporally with independent pulsed laser sources. We find that asphaltene nanoaggregates can be detected in LDI–MS and SALDI–MS under mild conditions of relatively low laser power, whereas L2MS avoids aggregation and fragmentation, detecting asphaltenes as monomeric molecules. A comparison of the L2MS and SALDI–MS results yields an estimate of the distribution of aggregation numbers (number of molecules comprising the nanoaggregate). The most probable aggregation number observed for nanoaggregates of petroleum asphaltenes is approximately 6–8 molecules, consistent with the Yen–Mullins model of asphaltenes and predictions from the island geometry for asphaltene molecules. Additionally, the nanoaggregates are found to be relatively monodisperse, because most aggregates observed have aggregation numbers within one molecule of the most probable. In contrast, the nanoaggregates of coal asphaltenes are found to be smaller and more polydisperse, with aggregation numbers ranging from 3 to 6 molecules. Under higher powers, SALDI–MS measurements show that nanoaggregates decompose to form small multimers and monomers, suggesting that the aggregates are bound noncovalently. Coal asphaltene nanoaggregates decompose at lower laser powers than petroleum asphaltene nanoaggregates, indicating that the coal asphaltene nanoaggregates are bound less strongly than petroleum asphaltene nanoaggregates.