Surface-assisted Laser Desorption Ionization Mass Research Articles

Serum lipidomic profiling for liver cancer screening using surface-assisted laser desorption ionization MS and machine learning

The liver is a major organ in metabolism, and alterations in serum lipids are associated with liver disorders. Here, a rapid, easy, and reliable screening technique based on lipidomic profiling was developed using machine learning and surface-assisted laser desorption ionization mass spectrometry (SALDI MS) for liver cancer diagnosis. A graphitized carbon matrix (GCM) was created for serum lipid profiling in SALDI MS and demonstrated a better performance for neutral lipids analysis than conventional organic matrices. The fingerprint of serum lipids, including triacylglycerols (TGs), diacylglycerols (DGs), cholesteryl esters (CEs), glycerophospholipids (GPs), and other components, could be directly obtained by GCM-assisted LDI MS without extraction. Five machine learning methods were applied to distinguish liver cancer (LC) patients from healthy controls (HC) and chronic hepatitis B (CHB) patients. The best diagnostic performance was attained by linear discriminant analysis (LDA), which has a confusion matrix accuracy of 98.3 %. The receiver operating characteristic (ROC) curve for liver cancer exhibited an area under the curve (AUC) of 0.99, indicating a high degree of prediction accuracy. One-way ANOVA analysis revealed that numerous TGs were down-regulated in LC group. The results demonstrated the viability of GCM-assisted LDI MS as a valuable diagnostic tool for liver cancer.

Aptamer-conjugated graphene oxide-based surface assisted laser desorption ionization mass spectrometry for selective extraction and detection of Aβ1-42 in an Alzheimer's disease SH-SY5 cell model.

The generation and accumulation of amyloid-beta peptide (Aβ1–42) in amyloid plaques are key characteristics of Alzheimer’s disease (AD); thus, specific detection of Aβ1–42 is essential for the diagnosis and treatment of AD. Herein, an aptamer-conjugated graphene oxide (Apt-GO) sensor was synthesized by π-π and hydrophobic interactions using thiol poly (ethylene glycol) amine (SH-PEG-NH2) as a spacer unit. Then, it was applied to selective capture of Aβ1–42, and the resulting complex was directly analyzed by surface-assisted laser desorption ionization mass spectrometry (SALDI-MS). The results revealed that the Apt-GO could enhance the detection specificity and reduce non-specific adsorption. This method was validated to be sensitive in detecting Aβ1–42 at a low level in human serum (ca. 0.1 μM) within a linear range from 0.1 to 10 μM. The immobilizing amount of aptamer on the GO was calculated to be 36.1 nmol/mg (RSD = 11.5%). In conclusion, this Apt-GO-based SALDI-MS method was sensitive and efficient in selective extraction and detection of Aβ1–42, which proved to be a good option for early AD diagnosis.

Nanostructured Layer of Silver for Detection of Small Biomolecules in Surface-Assisted Laser Desorption Ionization Mass Spectrometry.

A facile approach for the synthesis of a silver nanostructured layer for application in surface-assisted laser desorption/ionization mass spectrometry of low-molecular-weight biomolecules was developed using electrochemical deposition. The deposition was carried out using the following silver salts: trifluoroacetate, acetate and nitrate, varying the voltage and time. The plate based on trifluoroacetate at 10 V for 15 min showed intense SALDI-MS responses for standards of various classes of compounds: fatty acids, cyclitols, saccharides and lipids at a concentration of 1 nmol/spot, with values of the signal-to-noise ratio ≥50. The values of the limit of detection were 0.71 µM for adonitol, 2.08 µM for glucose and 0.39 µM for palmitic acid per spot. SEM analysis of the plate showed anisotropic flower-like microstructures with nanostructures on their surface. The reduced chemical background in the low-mass region can probably be explained by the absence of stabilizers and reducing agents during the synthesis. The plate synthesized with the developed approach showed potential for future use in the analysis of low-molecular-weight compounds of biological relevance. The absence of the need for the utilization of sophisticated equipment and the synthesis time (10 min) may benefit large-scale applications of the layer for the detection of various types of small biomolecules.

Dual-Mechanism-Driven Strategy for High-Coverage Detection of Serum Lipids on a Novel SALDI-MS Target.

Serum lipid metabolites have been emerging as ideal biomarkers for disease diagnosis and prediction. In the current stage, nontargeted or targeted lipidomic research mainly relies on a liquid chromatography-mass spectrometry (LC-MS) platform, but future clinical applications need more robust and high-speed platforms. Surface-assisted laser desorption ionization mass spectrometry (SALDI-MS) has shown excellent advantages in the high-speed analysis of lipid metabolites. However, the platform in the positive ion mode is more inclined to target a certain class of lipids, leading to the low coverage of lipid detection and limiting its practical translation to clinical applications. Herein, we proposed a dual-mechanism-driven strategy for high-coverage detection of serum lipids on a novel SALDI-MS target, which is a composite nanostructure comprising vertical silicon nanowires (VSiNWs) decorated with AuNPs and polydopamine (VSiNW-Au-PDA). The performance of laser desorption and ionization on the target can be enhanced by charge-driven desorption coupled with thermal-driven desorption. Simultaneous detection of 236 serum lipids (S/N ≥ 5) including neutral and polar lipids can be achieved in the positive ion mode. Among these, 107 lipid peaks were successfully identified. When combined with VSiNW-Au-PDA and VSiNW chips, 479 lipid peaks can be detected in serum samples in positive and negative ion modes, respectively. Based on the platform, serum samples from 57 hepatocellular carcinoma (HCC) patients and 76 healthy controls were analyzed. After data mining, 14 lipids containing different lipid types (TAG, CE, PC) were selected as potential lipidomic biomarkers. With the assistance of an artificial neural network, a diagnostic model with a sensitivity of 92.7% and a specificity of 96% was constructed for HCC diagnosis.

Laser Desorption Ionization–Mass Spectrometry with Graphite Carbon Black Nanoparticles for Simultaneous Detection of Taste- and Odor-Active Compounds

There is no analytical system to directly and simultaneously detect taste- and odor-active compounds. Graphite carbon black (GCB) nanoparticles, a chemical adsorbent for a variety of compounds, including gaseous molecules, which consists of nanoparticles made from multilayered sheets of sp2-conjugated atomic carbon, have the characteristics required for a surface-assisted laser desorption ionization–mass spectrometry (SALDI-MS) material. Thus, in this study, GCB-LDI-MS was developed for the simultaneous detection of taste- and odor-active compounds. The proposed GCB-LDI-MS successfully detected volatile compounds, including ethyl esters, alcohols, fatty acids, and aldehydes with up to 16 carbons (maximum carbon number of the tested compounds), which could not be detected by conventional matrix-assisted-LDI-MS. Moreover, a series of taste-active compounds (e.g., amino acids and sugars) and various volatile compounds (e.g., ethyl esters, aldehydes, alcohols, fatty acids, and lactones) as a mixture in liquid samples were also detected by the present GCB-LDI-MS. Thus, the proposed simple and rapid detection technique can digitize the analysis of flavor compounds, which provides an analytical concept for flavor scanning to evaluate the palatability of food products.

Detection of Acetaminophen and Its Glucuronide in Fingerprint by SALDI Mass Spectrometry Using Zeolite and Study of Time-Dependent Changes in Detected Ion Amount

The ingredients of an antipyretic (acetaminophen, AAP) and their metabolites excreted into fingerprint were detected by surface-assisted laser desorption ionization (SALDI) mass spectrometry using zeolite. In the fingerprint taken 4 h after AAP ingestion, not only AAP but also the glucuronic acid conjugate of AAP (GAAP), caffeine (Caf), ethenzamide (Eth), salicylamide (Sala; a metabolite of Eth), and urea were detected. Fingerprints were collected over time to determine how the amounts of AAP and its metabolite changed with time, and the time dependence of the peak intensities of protonated AAP and GAAP was measured. It was found that the increase of [GAAP+H]+ peak started later than that of [AAP+H]+ peak, reflecting the metabolism of AAP. Both AAP and GAAP reached maximum concentrations approximately 3 h after ingestion, and were excreted from the body with a half-life of approximately 3.3 h. In addition, fingerprint preservation was confirmed by optical microscopy, and fingerprint shape was retained even after laser irradiation of the fingerprint. Our method may be used in fingerprint analysis.

Application of Graphdiyne in Surface-Assisted Laser Desorption Ionization Mass Spectrometry.

Graphdiyne (GD) is a new kind of carbon nanomaterial which has carbon-carbon triple bonds to form a layered structure. Here, we report the application of GD as the matrix for small molecule analysis in laser desorption ionization mass spectrometry (LDI MS). The GD matrix displayed two advantages: little background in the low mass range and good molecular ion signal in negative ion mode for many small molecules, e.g., fatty acids, amino acids, peptides, and drugs can be obtained in negative ion mode. By comparing the signal intensity of tetraphenylborate and juglone with and without GD existing, it was found that GD can enhance both of the desorption efficiency and ionization efficiency in LDI. Through analysis of the serum samples from liver cancer patients and healthy people, the GD-assisted LDI MS results showed that fatty acids could be used as potential biomarkers for the early diagnosis of liver cancer.

Rapid Fingerprinting of High-Molecular-Weight Polymers by Laser Desorption-Ionization Using Through-Hole Alumina Membrane High-Resolution Mass Spectrometry

Residual acid found in the desorption ionization using through-holes alumina membranes (DIUTHAME) induces a reproducible protonation/in-source dissociation of polymers made of ester, amide, or siloxane moieties during their surface-assisted laser desorption ionization (SALDI) mass analysis. Deposited on the DIUTHAME chips in solution (solvent-based) or in pure form by melting the polymer powder in situ (solvent-free), high-molecular-weight nylons, silicone, or functionalized celluloses among other polymers are instantly fingerprinted by laser DIUTHAME high-resolution mass spectrometry (MS) with specific patterns resembling their direct analysis in real-time (DART) single-stage or tandem mass spectra. Depending on the polymer, two main types of fingerprints are observed with either the protonated monomer or product ions revealing the nature of the repeating unit or its functionalization. This technique allows a rapid molecular analysis of industrial homopolymers regardless of their molecular weight and complementary to DART with simple or no sample preparation and also promisingly applicable for copolymers.

Crystalline MOF nanofilm-based SALDI-MS array for determination of small molecules.

A highly oriented crystalline metal-organic framework (MOF) nanofilm array was prepared and used for surface-assisted laser desorption ionization mass spectrometry (SALDI-MS) for determinationof small molecules. The MOF nanofilm was characterized using scanning/transmission electron microscopy, X-ray diffraction, and ultraviolet-visible spectroscopy. Different small molecules (anthracene, n-eicosanoic acid, Rhodamine B) were successfully determined by this MOF nanofilm array with limits of detection (LOD) between 0.1-5ng·mL-1 and limits of quantification (LOQ) between 1 and 10ng·mL-1. Compared topreviously reported MOF-based SALDI-MS, this array exhibits better reproducibility (3.3-5.2%) and recovery (89-105%). The intensity of the MS peak remains the same after 25 repeated cycles. This indicates good repeatability. This MOF nanofilm-based SALDI-MS array can be used for determination of fatty acid and Rhodamine B in real samples with good recovery (83-106%). Graphical abstract Schematic representation of the principle of crystalline MOFs nanofilm-based SALDI-MS array.

Gas-aggregated Ag nanoparticles for detection of small molecules using LDI MS.

Several reports demonstrate that silver nanomaterials can serve as surface-assisted laser desorption ionization mass spectrometry (SALDI MS) substrates for low molecular weight analytes. Substrate with tailored silver nanostructures, primarily representing the upmost layer of the bulk, i.e., occurring beneath the analyzed medium, limits the use of silver only for desorption enhancement; the charge transfer progresses through atoms from the absorbing analyte or an additional matrix (resulting in the formation of analyte/hydrogen, sodium, or potassium adducts in the most cases). In the presented approach, we utilize a homogeneous layer of silver nanoparticles, prepared under low-pressure conditions, deposited onto a dried analyte. We demonstrate that the nanoparticle layer can fully replace a matrix typically used for the detection of small molecules by laser desorption/ionization mass spectrometry-based technique (LDI MS) and can be applied to the already prepared samples. Various chemical species were detected as [M + Ag]+ adduct ions employing the proposed technique. The normalized signal of the analyte/silver adduct can be utilized to characterize a quantitative presence of analytes on the surface similar to signal-to-noise value, here demonstrated by the detection of trimethoprim molecule. This study also includes a detailed description of additional features one needs to take into account, such as a formation of [Mx+ Agy]+ adducts, presence of silver ions (can be used for m/z calibration), analyte fragmentation, and influence of deposited nanoparticles quantity on the signal intensity. Graphical abstract.

Front Cover: TiO2 Nanocoral Structures as Versatile Substrates for Surface‐Assisted Laser Desorption/Ionization Mass Spectrometry (ChemNanoMat 4/2019)

The cover image illustrates the matrix-free surface-assisted laser desorption/ionization (SALDI) of small organic molecules, gold clusters, and polyoxometalates using versatile TiO2 nanocoral (TNC) substrates. SALDI mass spectrometry (MS) is a powerful technique for rapidly analyzing a myriad of compounds including biomolecules and polymers. However, previous studies were limited to detecting natural/organic-based molecules. Biphasic TNC structures consisting of an anatase base film and a rutile nanocoral layer allow analysis of not only organic compounds and polymers but also inorganic clusters by SALDI-MS. More information can be found in the Full Paper by Tetsuro Soejima et al. on page 447 in Issue 4, 2019 (DOI: 10.1002/cnma.201800588).

TiO2 Nanocoral Structures as Versatile Substrates for Surface‐Assisted Laser Desorption/Ionization Mass Spectrometry

AbstractMatrix‐free surface‐assisted laser desorption/ionization (SALDI) is a powerful technique for rapidly analyzing a myriad of compounds including biomolecules and polymers. However, previous studies were limited to detecting natural/organic‐based molecules. Biphasic TiO2 nanocoral (TNC) structures consisting of an anatase base film and a rutile nanocoral layer allow analysis of not only organic compounds and polymers but also inorganic clusters by SALDI mass spectrometry (MS). TNC structures with microscopic surface roughness and porosity, which are indispensable for efficient desorption/ionization (D/I) in SALDI, form on Si wafers immersed in aqueous TiOSO4‐H2O2 at 80 °C. Various organic and biological molecules with a wide range of molecular weights such as amino acids, hormones, antioxidants, steroids, organic dyes, sugars, drugs, proteins, and polymers are successfully analyzed by using TNC substrates in SALDI‐MS. After SALDI‐MS, the TNC substrates can be reused for detecting water‐soluble molecules. TNC substrates also allow efficient detection of polyoxometalates and gold clusters as representative inorganic clusters. Survival yield measurements are employed to investigate the D/I mechanism of TNC substrates, and these indicate that the rutile nanocoral layer plays a critical role in ensuring efficient D/I. TNC as a versatile substrate for SALDI‐MS is a potentially important analysis tool in a wide range of chemical and biological fields.

Enhancing surface-assisted laser desorption ionization mass spectrometry performance by integrating plasmonic hot-electron transfer effect through surface modification.

Surface-assisted laser desorption ionization mass spectrometry (SALDI MS) performances were enhanced by modifying surfaces to include a plasmonic hot-electron transfer effect. After modification, the core material's specificity was not only maintained but also enhanced. The MS imaging of ischemic tissues with the modified material indicated some sulfatide changes, which have been barely reported.

Silylation with the Generation of a Fixed Charge for the Analysis of Alcohols by MALDI and SALDI Mass Spectrometry

A new method of the derivatization of alcohols for their analysis by matrix-assisted laser desorption/ionization (MALDI) and surface-assisted laser desorption/ionization (SALDI) is proposed. The approach is based on the silylation of the analytes with bromomethylchlorodimethylsilane in the presence of tertiary amines to yield fixed-charge containing derivatives. The proposed method was tested using aliphatic alcohols, terpenols, and sterols. MALDI and SALDI mass spectra of all derivatives contain intense ion peaks corresponding to their cationic parts.

Small- and large-sized iron(II, III) oxide nanoparticles for surface-assisted laser desorption/ionization mass spectrometry of small biomolecules.

Common surface-assisted laser desorption/ionization (SALDI) surfaces are functionalized to improve mass spectrometric detection. Such surfaces are selective to certain group(s) of compounds. The application of universal and sensitive SALDI surfaces with appropriate size/surface area is paramount. In this study, two different sizes/surface areas of Fe3 O4 are compared as SALDI surfaces. For accurate surface area comparisons, the physical properties of the Fe3 O4 nanoparticles used as SALDI surfaces were determined using scanning electron microscopy, X-ray diffractometry, and N2 Brunauer-Emmet-Teller adsorption techniques. SALDI mass spectrometry (MS) data were acquired using a time-of-flight (TOF) mass spectrometer operated in the linear mode and equipped with a 50-Hz pulsed nitrogen laser (at 337 nm). Small biomolecules (adenosine, glucose, sucrose, tryptophan, and tripeptide) and a real sample (human serum) were analyzed. The average sizes/specific surface areas of the SALDI surfaces of the small- and large-sized Fe3 O4 nanoparticles were ~21 nm/~82 m2 /g and ~39 nm/~38 m2 /g, respectively. An overall ~2.0-fold enhancement in signal-to-noise ratios was observed for the ionic species of the analyzed biomolecules in SALDI-MS using small-sized Fe3 O4 in comparison to large-sized Fe3 O4 nanoparticles. MS sensitivity from adenosine calibration curves (concentration between 0.05 and 10.0 mM) was ~2.0-fold higher for small-sized than large-sized Fe3 O4 nanoparticles as SALDI surfaces. We have shown that transition-metal oxides such as Fe3 O4 nanoparticles are suitable and efficient surfaces for SALDI-TOF-MS analysis of small biomolecules. We observed improvement in signal-to-noise ratios and detection sensitivity for the analyzed samples from SALDI surfaces using small-sized (possessing larger surface area) than large-sized Fe3 O4 nanoparticles.

Chip-based SALDI-MS for rapid determination of intracellular ratios of glutathione to glutathione disulfide

Alterations in the ratio of glutathione (GSH) to glutathione disulfide (GSSG) reveal the cell living state and are associated with a variety of diseases. In this study, an Au NPs grafted nanoporous silicon chip was used for surface assisted laser desorption ionization-mass spectrometry (SALDI-MS) detection of GSH. Due to the bond interaction between thiol of GSH and Au NPs modified on the chip surfaces, GSH could be captured from the complex cellular lysate. Meanwhile, the composite nanostructures of Au NPs grafted porous silicon surface presented good desorption/ionization efficiency for GSH detection. The GSH levels in different tumor cells were successfully detected. Chip-based SALDI-MS was optimized for quantification of intracellular GSH/GSSG ratio changing under drug stimulation in liver tumor cells, GSSG was reduced to GSH by reductant of tris(2-carboxyethyl)phosphine (TCEP) and isotope-labeling GSH was as an internal standard. It was found that the increasing concentration of drug irinotecan and hypoxia culture condition caused the rapid consumption of GSH and a decrease of GSH/GSSG ratio in liver tumor cells. The developed SALDI-MS method provided a convenient way to accurately measure and rapidly monitor cellular GSH value and the ratios of GSH/GSSG.

The Cooperativity of Fe3 O4 and Metal-Organic Framework as Multifunctional Nanocomposites for Laser Desorption Ionization Process.

A novel and facile strategy in developing a water stable magnetic metal-organic framework nanocomposite (Fe3 O4 @MOF) is herein reported, in which a Keggin polyoxometalate, phosphotungstic acid (HPW), was encapsulated within the MOF framework via one-pot synthesis method. The combination of HPW-embedded MOF and Fe3 O4 endowed the composite with high surface area, strong UV absorption, good hydrophilicity, and enhanced water stability. With these unique properties, the Fe3 O4 @MOF embedded HPW served as adsorbent as well as matrix for SALDI-MS (surface-assisted laser desorption ionization mass spectrometry) analysis of polar and non-polar compounds. The synergistic effect of Fe3 O4 and MOF showed an interference-free background at low mass region than the pristine MOF or Fe3 O4 counterparts. This simple approach can be used as new platform in developing magnetic MOF composites without the time consuming and labor-intensive preparation.

Nanoaggregates of Diverse Asphaltenes by Mass Spectrometry and Molecular Dynamics

Asphaltene nanoaggregates for three diverse source materials—coal-derived asphaltenes dominated by aromatic carbon, petroleum asphaltenes with comparable abundances of aromatic and aliphatic carbon, and immature source-rock asphaltenes dominated by aliphatic carbon—are examined by means of surface-assisted laser desorption ionization mass spectrometer (SALDI-MS) coupled with laser desorption laser ionization mass spectrometry (L2MS). All three types of asphaltenes form nanoaggregates with aggregation numbers close to 7. Molecular dynamics calculations for proposed island molecular structures show the important roles that π-stacking and alkane steric hindrance play in nanoaggregate formation. These results are discussed in terms of entropy and enthalpy changes. All results are consistent with the Yen-Mullins model, which bodes well for its expanded use in oilfield reservoir evaluations.

Carbon nanocoatings: A new approach to recording mass spectra of low-molecular compounds using surface-assisted laser desorption/ionization mass spectrometry

A simple and rapid approach to obtaining target plates for the investigation of low-molecularweight compounds by surface-assisted laser desorption/ionization (SALDI) mass spectrometry is proposed. It consists in the vacuum sputtering of a carbon layer with a thickness of about 50 nm onto a metal surface. The resulting coatings are characterized by homogeneity, hydrophobicity, and high mechanical strength, which eliminates a possibility of mass spectrometer contamination. A comparison of the SALDI mass spectra of test compounds recorded using conventional carbon materials and carbon nanocoatings demonstrates advantages of the last named materials, such as high spectral resolution and the absence of spectral interferences at low m/z values.

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.