High-resolution TOF-SIMS study of varying chain length self-assembled monolayer surfaces

A high-resolution time-of-flight secondary ionization mass spectrometer (TOF-SIMS) has been used to investigate chain length effects in hydrocarbon seff-assembled monolayer (SAM) surfaces on gold substrates. A wide range of n-alkanethiols was used to make homogeneous SAM surfaces, which included both odd and even hydrocarbon chain length thiols. Variations in coverage, extent of oxidation, and high-mass cluster formation as a function of hydrocarbon chain length of the alkanethiol SAM surfaces were investigated. Long-short chain length effects were observed for the relative coverage of the SAM surfaces, which directly influences the extent of oxidation for the thin films. The formation of gold-sulfur and gold-adsorbate cluster ions was also observed, since the mass range of the TOF-SIMS made it possible to monitor all of the cluster ions that were formed following the high-energy ion/surface interactions.     

Scrubbing ions with molecules: kinetic studies of chemical noise reduction in mass spectrometry using ion-molecule reactions with dimethyl disulfide

The kinetics and product distributions of the reactions of dimethyl disulfide (DMDS) have been investigated with a group of chemical background ions commonly observed in atmospheric pressure ionization (API) mass spectrometry (MS) in order to assess the value of this molecule in filtering (or "scrubbing") these ions by changing their mass/charge (m/z) ratio. The measurements were taken with a novel electrospray ionization/selected ion flow tube/QqQ tandem mass spectrometer. The background ions studied include those with m/z 42 (protonated acetonitrile, ACN), 83 (protonated ACN dimer), 99 (protonated phosphoric acid), 117 (water cluster of m/z 99), 131 (methanol cluster of m/z 99), 149 (protonated phthalic anhydride, formed from the phthalates), and 327 (protonated triphenyl phosphate). In addition, reactions of DMDS have been studied with two model analytes--protonated caffeine and doubly protonated bradykinin--in order to assess the selectivity of DMDS reactivity. All the measurements were taken at 295 +/- 2 K in helium buffer gas at a pressure of 0.35 +/- 0.01 Torr. DMDS was observed to react efficiently with m/z 42 (ACNH+), 149 (from phthalates), and 99 (protonated phosphoric acid), with k/kc=0.91, 0.47, and 0.38, respectively. Only proton transfer was observed with ACNH+, followed by the secondary reaction of [DMDSH]+ with DMDS to yield [CH3S-S(CH3)-SCH3]+. Ligation of DMDS was the dominant primary channel observed for the reaction of the m/z 149 background ion; however, some proton transfer also was observed. Both of these primary product ions react further with DMDS to yield [CH3S-S(CH3)-SCH3]+, the structure of which we have determined computationally using DFT calculations. Only the sequential ligation with two DMDS molecules was observed for the reaction of the m/z 99 ion. Reactions of DMDS with m/z 117 [H3PO4 + H + H2O]+ and m/z 131 [H3PO4 + H + MeOH]+ were observed to proceed with k/kc=0.71 and 0.058, respectively. Ligand substitution of DMDS for H2O predominated ( approximately 94%) over DMDS ligation ( approximately 6%) in the reaction with m/z 117, while only DMDS ligation was observed for the reaction of m/z 131 with DMDS. In contrast, the reactions of DMDS with ions of m/z 83 (protonated dimer of ACN) and 327 (protonated triphenyl phosphate) were extremely inefficient, with k/kc=0.0042 and 0.0079, respectively. The higher reactivity of DMDS toward ACNH+ (m/z 42) compared to (ACN)2H+ (m/z 83) is attributed to the lower proton affinity of the unsolvated ACN. The reactivity of DMDS toward the two model analyte ions studied-protonated caffeine and doubly protonated bradykinin-was negligible, with k/kc=0.0073 and 0.010, for the respective reactions. These results suggest that, under appropriate reagent pressure conditions, DMDS can be an appropriate reagent for chemically filtering out many common API-MS background ions, without significantly affecting the observed intensity of analyte peaks.     

Comprehensive assignment of mass spectral signatures from individual Bacillus atrophaeus spores in matrix-free laser desorption/ionization bioaerosol mass spectrometry

We have fully characterized the mass spectral signatures of individual Bacillus atrophaeus spores obtained using matrix-free laser desorption/ionization bioaerosol mass spectrometry (BAMS). Mass spectra of spores grown in unlabeled, 13C-labeled, and 15N-labeled growth media were used to determine the number of carbon and nitrogen atoms associated with each mass peak observed in mass spectra from positive and negative ions. To determine the parent ion structure associated with fragment ion peaks, the fragmentation patterns of several chemical standards were independently determined. Our results confirm prior assignments of dipicolinic acid, amino acids, and calcium complex ions made in the spore mass spectra. The identities of several previously unidentified mass peaks, key to the recognition of Bacillus spores by BAMS, have also been revealed. Specifically, a set of fragment peaks in the negative polarity is shown to be consistent with the fragmentation pattern of purine nucleobase-containing compounds. The identity of m/z = +74, a marker peak that helps discriminate B. atrophaeus from Bacillus thuringiensis spores grown in rich media is [N1C4H12]+. A probable precursor molecule for the [N1C4H12]+ ion observed in spore spectra is trimethylglycine (+N(CH3)3CH2COOH), which produces a m/z = +74 peak when ionized in the presence of dipicolinic acid. A clear assignment of all the mass peaks in the spectra from bacterial spores, as presented in this work, establishes their relationship to the spore chemical composition and facilitates the evaluation of the robustness of "marker" peaks. This is especially relevant for peaks that have been used to discriminate Bacillus spore species, B. thuringiensis and B. atrophaeus, in our previous studies.     

Molecular dynamics simulation of the enhancement of cobra cardiotoxin and E6 protein binding on mixed self-assembled monolayer molecules

Molecular dynamics simulations are performed on n-alkinethiol self-assembled monolayers (SAMs) and their mixture on a gold surface so that the orientations of the binding of cobra cardiotoxin and E6 protein molecules can be selected using the mixing ratio of CH3-terminated SAMs with different chain lengths. The simulations suggest that a SAM surface with different mixing ratios may provide a possible platform for aligning protein molecules with a desired orientation and for enhancing the binding energy of the protein on the designed surface.     

Characterization of photooxidized self-assembled monolayers and bilayers by spontaneous desorption mass spectrometry

We show that selected self-assembled monolayers (SAMs) and bilayers are readily characterized by the application of controlled photooxidation and spontaneous desorption mass spectrometry (SDMS) in the negative ion mode. Additionally, SDMS is used to characterize organic and inorganic anionic species adsorbed to the surface of a positively charged SAM surface, 2-aminoethanethiol (AET). Prominent peaks are observed that correspond both to the sulfonate form of each SAM and bilayer and to the anion form of each molecule adsorbed to AET. In addition, fragments of the oxidized thin films were also observed at m/z 80 (SO3-) and 97 (HSO4-). Other prominent fragment peaks more characteristic of the molecule are also seen in the mass spectra.     

Sequencing of argentinated peptides by means of electrospray tandem mass spectrometry.

A strategy for semiautomatic sequencing of argentinated (silver-containing) oligopeptides has been developed. Sequencing is based on a search algorithm that identifies a triplet peak relationship in a product ion spectrum of the [M + Ag]+ ion of an oligopeptide. The ions that constitute a triplet are [bn + OH + Ag]+, [bn - H + Ag]+, and [a(n) - H + Ag]+, which are separated by 18 and 28 m/z units, respectively. The difference in the m/z values of adjacent triplets identifies the residue that is "cleaved". Observation of the [yn + H + Ag]+ ion containing the cleaved residue confirms the assignment. Sequencing of argentinated tryptic peptides may prove useful for automated proteome analysis via the sequence tag method.     

Palladium as a substrate for self-assembled monolayers used in biotechnology

This paper describes self-assembled monolayers (SAMs) on palladium that resist the nonspecific adsorption of proteins and the adhesion of mammalian cells. These SAMs form when thin films of palladium are exposed to solutions of alkanethiol with the general structure HS(CH(2))(m)()(OCH(2)CH(2))(n)()OH (m = 2, 11; n = 3, 6, 7). Ellipsometry and surface plasmon resonance spectroscopy (using a palladium-on-gold substrate) showed that these SAMs resist adsorption of all proteins present in bovine serum. Microislands of SAMs of octadecanethiol on palladium allowed patterned adhesion and growth of mammalian cells (in a "sea" of oligo(ethyleneglycol)-terminated SAM). The oligo(ethyleneglycol)-terminated SAM resisted the invasion of cells for over four weeks under standard conditions of cell culture; similar SAMs on gold remained patterned for only two weeks.     

Atmospheric pressure covalent adduct chemical ionization tandem mass spectrometry for double bond localization in monoene- and diene-containing triacylglycerols

We report a method to elucidate the structure of triacyl-glycerols (TAGs) containing monoene or diene fatty acyl groups by atmospheric pressure covalent adduct chemical ionization (APCACI) tandem mass spectrometry using acetonitrile as an adduct formation reagent. TAGs were synthesized with the structures ABB and BAB, where A is palmitate (C16:0) and B is an isomeric C18 monoene unsaturated at position 9, 11, or 13 or an isomeric diene unsaturated at positions 9 and 11, 10 and 12, or 9 and 12. In addition to the species at m/z 54 observed in previous CI studies of fatty acid methyl esters, we also found that ions at m/z 42, 81, and 95 undergo covalent reaction with TAGs containing double bonds to yield ions at m/z 40, 54, 81, and 95 units greater than that of the parent TAG: [M + 40]+, [M + 54]+, [M + 81]+, and [M + 95]+ ions. When collisionally dissociated, these ions fragment to produce two or three diagnostic ions that locate the double bonds in the TAG. In addition, ions [RCH=C=O + 40]+ and [RCH=C=O + 54]+ formed from collisional dissociation are of strong abundance in MS/MS spectra, and collisional activation of these ions produces two intense confirmatory diagnostic ions in the MS3 spectra. Fragment ions reflecting neutral loss of an sn-1-acyl group from [M + 40]+ and [M + 54]+ are more abundant than those reflecting neutral loss of an sn-2-acyl group, analogous to previous reports for protonated TAGs. The position of each acyl group on the glycerol backbone is thus determined by the relative abundances of these ions. Under the conditions in our instrument, the [M + 40]+ adduct is at the highest signal and also yields all information about the double bond position and TAG stereochemistry. With the exception of geometries about the double bonds, racemic TAG isomers containing two monoenes or dienes and a saturate can be fully characterized by APCACI-MS/MS/MS.     

Molecular interactions in self-assembly monolayers on gold-coated microcantilever electrodes

An electrochemical microcantilever (EMC) was used to study the intermolecular interaction of self-assembly monolayers (SAMs) with different n-alkanethiols chain lengths (n = 0, 4, 6, 8, 12, 16) on a Au-coated microcantilever surface. Comparing potential cycling and steps in NaClO(4) solution within the same potential range, the deflection rate of bare microcantilevers is much smaller for the former which revealed that potential excitation, i.e. the surface charge, played the dominant role in driving the instant and large deflection of the bare microcantilever, while the smaller deflection amplitude of the former implied that adsorption of ClO(4)( - ) had an adverse effect on the potential-induced stress. Upon adsorption of SAMs, the deflection amplitude of the microcantilever under the potential step was much smaller than that of a bare microcantilever, and linearly decreased with the chain length increasing for n ≤ 8 (the linear correlation coefficient and the slope are 0.98 and about - 10.4 nm per CH(2) unit, respectively), following a transition (8 ≤ n ≤ 12) to a stable state (n ≥ 12). The decrease of deflection amplitude and faster decay of deflection rate of the SAMs modified microcantilever under the potential step implyed increasing compactness of the SAMs with longer chains.     

微波等离子体化学气相沉积法生长非晶碳化硅薄膜

利用SiH4(80%Ar稀释)和CH4作为源气体,通过改变源气体流量比、基片温度、沉积气压等参量,使用微波电子回旋共振化学气相沉积法生长非晶碳化硅薄膜。实验结果表明碳化硅薄膜沉积速率随气体流量比R(CH4/(CH4+SiH4))的增加而减小、随基片温度的升高明显减小、随沉积气压的增加先增大后减小。红外结构表明:在较低流量比R下,薄膜主要由硅团簇和非晶碳化硅两相组成,而当R>0.5时,薄膜的结构主要由非晶碳化硅组成,薄膜中键合的H主要是Si和C的封端原子。同时,沉积温度的升高使碳化硅薄膜中Si-H,C-C和C-H键的含量减少,而薄膜中Si-C含量明显增加且峰位发生了红移。薄膜相结构的转变是薄膜光学带隙变化的原因。     

Tandem mass spectrometry characteristics of silver-cationized polystyrenes: backbone degradation via free radical chemistry

The [M + Ag](+) ions of polystyrene (PS) oligomers are formed by matrix-assisted laser desorption/ionization, and their fragmentation characteristics are determined by tandem mass spectrometry experiments in a quadrupole/time-of-flight mass spectrometer. Collisionally activated dissociation (CAD) of [M + Ag](+) starts with random homolytic C-C bond cleavages in the PS chain, which generate radical ions carrying either the initiating (a(n*), b(n*)) or the terminating (y(n*), z(n*)) chain end and primary (a(n*), y(n*)) or benzylic (b(n*), z(n*)) radical centers. The fragments ultimately observed arise by consecutive, radical-induced dissociations. The primary radical ions mainly decompose by monomer evaporation and, to a lesser extent, by beta-H(*) loss. The benzylic radical ions primarily decompose by 1,5-H rearrangement (backbiting) followed by beta C-C bond scissions; this pathway leads to either closed-shell fragments with CH(2) end groups, internal fragments with 2-3 repeat units, or truncated benzylic b(n*)/z(n*) radical ions that can undergo anew backbiting. The same internal fragments are produced in all backbiting steps; hence, these fragments and small benzylic radical ions (which cannot undergo backbiting) dominate the low-mass region of the CAD spectra, while the less abundant closed-shell fragments with CH(2) end groups (a(n)/y(n)) dominate the medium- and high-mass regions. The latter fragments are suitable for determining the individual initiating and terminating end groups, whereas the internal ions could be valuable in sequence analyses of styrene copolymers.     

Identification of microcystin toxins from a strain of Microcystis aeruginosa by liquid chromatography introduction into a hybrid linear ion trap-Fourier transform ion cyclotron resonance mass spectrometer

The cyclic heptapeptide microcystin toxins produced by a strain of Microcystis aeruginosa that has not been investigated previously were separated by liquid chromatography and identified by high-accuracy m/z measurements of their [M + H]+ ions and the fragment ions produced by collision-activated dissociation of the [M + H]+ ions. The cyanobacteria B2666 strain was cultured in a standard growth medium, and the toxins were released from the cells, extracted from the aqueous phase, and concentrated using standard procedures. The microcystins were separated by reversed-phase microbore liquid chromatography and introduced directly into a hybrid linear ion trap-Fourier transform ion cyclotron resonance mass spectrometer with electrospray ionization. The known microcystins (MC) MC-LR, MC-LA, [MeSer7]MC-LR, MC-LL, MC-LF, and MC-L(Aba) were identified along with the two previously unreported structural variants [Asp3]MC-LA and [Asp3]MC-LL. In addition to the [M + H]+ ions, accurate m/z measurements were made of 12-18 product ions for each identified microcystin. The mean difference between measured and calculated exact m/z was less than 2 parts per million, which often allowed assignment of unique compositions to the observed ions. A mechanism is presented that accounts for an important collision-activated dissociation process that gives valuable sequence ions from microcystins that do not contain arginine. The analytical technique used in this work is capable of supporting fairly rapid and very reliable identifications of known microcystins when standards are not available and of most structural variants independent of additional information from other analytical techniques.     

Dicationic ion-pairing agents for the mass spectrometric determination of perchlorate

Perchlorate and other hydrophobic ions can be measured with high sensitivity and selectivity by forming a positively charged ion pair with a dicationic agent. A commercially available reagent, 1,6-bis(trimethylammonium)hexane dibromide (Br(N(CH3)3)(CH2)6(N(CH3)3)Br) allows for the determination of perchlorate by electrospray ionization mass spectrometry as the [(N(CH3)3)(CH2)6(N(CH3)3)ClO4]+ ion. Limits of detection (LODs) are better than those previously observed with custom-synthesized dicationic agents. An LOD of 20 ng/L is readily attainable with a single-quadrupole mass spectrometer.     

高效液相色谱-串联质谱法测定贻贝中腹泻性贝类毒素的含量

采用液相色谱-串联质谱法检测了贻贝中大田软海绵酸(OA)和鳍藻毒素-1(DTX-1)两种腹泻性贝类毒素的含量。样品经80%甲醇水溶液提取,Sep-pak silica固相萃取小柱净化,80%甲醇水溶液定容后供HPLC-MS/MS分析。采用电喷雾负离子模式多反应监测方式进行检测,OA和DTX-1的定量检测的离子对分别为m/z 803.5/255.1和m/z817.4/255.1。2种贝类毒素在20~800μg/L范围内线性良好;在4个添加水平下OA的回收率为79.5%～88.6%,RSD为8.43%～10.4%;DTX-1的回收率为83.8%～91.2%,RSD为4.22%～6.54%。方法灵敏度高,定量限为0.02mg/kg。来自市场和产地的45个贻贝样品残留分析发现,有4个样品检出腹泻性贝类毒素,检出率为8.9%。     

Parent and neutral loss monitoring on a quadrupole ion trap mass spectrometer: screening of acylcarnitines in complex mixtures

A novel and practical technique for performing both parent and neutral loss (P&NL) monitoring experiments on a quadrupole ion trap mass spectrometer is presented. This technique is capable of performing scans analogous to the parent and neutral loss scans routinely applied on tandem-in-space instruments and allows for the screening of a sample to detect analytes of a specific compound class on a chromatographic time-scale. Acylcarnitines were chosen as the model compound class to demonstrate the analytical utility of P&NL monitoring because of their amenability to electrospray ionization (ESI), their unique and informative MS/MS fragmentation pattern, and their importance in biological functions. The [M + H]+ ions of all acylcarnitines dissociate to produce neutral losses of 59 and 161 amu and common product ions at m/z 60, 85, and 144. Both the neutral loss monitoring of 59 amu and the parent ion monitoring of m/z 85 are shown to be capable of identifying acylcarnitine [M + H]+ ions in a synthetic mixture and spiked pig plasma. The neutral loss monitoring of 59 amu is successful in detecting acylcarnitines in an unspiked pig plasma sample.     

Mass spectrometric analysis for high molecular weight synthetic polymers using ultrasonic degradation and the mechanism of degradation

We have investigated ultrasonic degradations of poly(ethylene oxide) (PEG) and poly(methyl methacrylate) (PMMA) in aqueous media by means of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The ultrasonic degradation of polymers was monitored as a function of ultrasonication duration to examine the structural details of ultrasonic degradation polymers. PEG solution ultrasonication produced five types of oligomers (M approximately 1000 Da) with different end groups, irrespective of the initial average molecular masses (M=2, 6, 20, and 2000 kDa). Several degradation pathways with free radical reactions have been suggested to explain these degradation products: the ultrasonic degradation of PEG is initiated by breaking of the C-O bond in the PEG chain, generating polymeric radicals with two terminal groups, i.e., X*( approximately CH2CH2*) and Y*( approximately CH2CH2O*), followed by termination with extraction or release of a hydrogen atom. However, PMMA (M=1630 Da) ultrasonication generated only one type of degradation oligomer, which has a hydrogen group at both ends, the same as that of the original oligomer. It has been suggested that the presence of the radical terminal groups X*( approximately CH2*) and Y*( approximately (CH3)CCOO(CH3)C*) is due to selective C-C bond breaking in the chain during the ultrasonic degradation of PMMA. The MALDI-TOFMS combined with the ultrasonic degradation technique (UD/MALDI-TOFMS) developed in this study could be extended to the analysis of synthetic polymer structures with high molecular weights.     

Characterization of C-N Thin Films Prepared by Ion Beam Sputtering

C-N thin films have been prepared by ion beam sputtering using pure N2 as discharge gas. The ratio N/C of the films measured by Auger spectrum is 20% on an average. The results of X-ray diffraction and transmission electron microscopy show that β-C3N4 phase exists in the films. Xray photoelectron spectroscopy shows that nitrogen is mostly combined with carbon with triple (C=N) and double (C=N) bonds. The IR absorption shows an absorption bond near 2185 cm-1assigned to the C=N, no trace of C-C bond was observed     

A validated liquid chromatography/tandem mass spectrometry assay for cis-amminedichloro(2-methylpyridine)platinum(II) in human plasma ultrafiltrate

The clinical use of platinum drugs as anticancer agents has encountered problems when relating pharmacokinetic profiles with efficacy and toxicity is attempted. This has been mainly due to the lack of specific and sensitive analytical methodology to examine concentrations of the unbound drug in plasma. The presence of a carbocyclic ring on the new drug, cis-amminedichloro(2-methylpyridine)platinum(II) (ZD0473) suggested that it would be possible to develop the first stable isotope dilution LC/MS assay for a platinum drug in human plasma ultrafiltrate samples. The dichloro form of the drug exists in equilibrium with at least two aquated forms in plasma. The molecular form of the drug, therefore, depends on the length of time that the plasma sample is maintained at room temperature before freezing. Therefore, we have developed a method that quantitatively converts the aquated species back to the dichloro form of the parent drug so that a single molecular species can be analyzed. Selected reaction monitoring was performed on the transition of m/z 393 [M + NH4]+ to m/z 304 [M + NH4 -NH3 - 2 x HCl]- for ZD0473, and m/z 400 [M + NH4]+ to m/z 310 [M + NH4 - NH3 - HCl - 2HCl]+ for [2H7]ZD0473. The standard curves were fitted to a quadratic regression over the range from 10 to 5000 ng/mL in human plasma ultrafiltrate. The lower limit of quantitation for ZD0473 was 10 ng/mL for 100 microL of plasma ultrafiltrate. This simple, rapid, reliable, and sensitive method of quantitation had excellent accuracy and precision. The method provided adequate sensitivity for the analysis of plasma ultrafiltrate samples from a phase II study in which ZD0473 was administered to patients as an intravenous infusion at a dose of 150 mg/m2.     

Analysis of intact tissue by intermediate-pressure MALDI on a linear ion trap mass spectrometer

The use of an intermediate-pressure matrix-assisted laser desorption/ionization (IP-MALDI) source working at 0.17 Torr on a linear ion trap (LIT) was investigated for the analysis of tissue specimens, in particular, spinal cord sections. MALDI, with 2,5-dihydroxybenzoic acid (DHB) as the matrix, was employed for the detection of phospholipids. The matrix was applied to the tissue using electrospray to avoid analyte migration. The results indicate that analyzing tissue specimens at nontraditional MALDI vacuum pressures is possible. Coupling MALDI to an LIT permits the use of MSn, which is critical for the ability to identify compounds desorbed directly from tissue specimens. Using MSn, ions detected from m/z 600-1000 were characterized as phosphatidlycholines, PC. Specifically, using tandem MS, PC ions could be classified as either [M + H]+ or [M + Na]+ because the fragmentation patterns of protonated and sodiated phosphatidlycholines follow different pathways.