Guangming Huang

New ionization methods and miniature mass spectrometers for biomedicine: DESI imaging for cancer diagnostics and paper spray ionization for therapeutic drug monitoring

The state-of-the-art in two new ambient ionization methods for mass spectrometry, desorption electrospray ionization (DESI) and paper spray (PS), is described and their utility is illustrated with new studies on tissue imaging and biofluid analysis. DESI is an ambient ionization method that can be performed on untreated histological sections of biological tissue in the open lab environment to image lipids, fatty acids, hormones and other compounds. Paper spray is performed in the open lab too; it involves electrospraying dry blood spots or biofluid deposits from a porous medium. PS is characterized by extreme simplicity and speed: a spot of whole blood or other biofluid is analyzed directly from paper, simply by applying a high voltage to the moist paper. Both methods are being developed for use in diagnostics as a means to inform therapy. DESI imaging is applied to create molecular maps of tissue sections without prior labeling or other sample preparation. Like other methods of mass spectrometry imaging (MSI), it combines the chemical speciation of multiple analytes with information on spatial distributions. DESI imaging provides valuable information which correlates with the disease state of tissue as determined by standard histochemical methods. Positive-ion data are presented which complement previously reported negative-ion data on paired human bladder cancerous and adjacent normal tissue sections from 20 patients. These data add to the evidence already in the literature demonstrating that differences in the distributions of particular lipids contain disease-diagnostic information. Multivariate statistical analysis using principal component analysis (PCA) is used to analyze the imaging MS data, and so confirm differences between the lipid profiles of diseased and healthy tissue types. As more such data is acquired, DESI imaging has the potential to be a diagnostic tool for future cancer detection in situ; this suggests a potential role in guiding therapy in parallel with standard histochemical and immunohistological methods. The PS methodology is aimed at high-throughput clinical measurement of quantitative levels of particular therapeutic agents in blood and other biofluids. The experiment allows individual drugs to be quantified at therapeutic levels and data is presented showing quantitative drug analysis from mixtures of therapeutic drugs in whole blood. Data on cholesterol sulfate, a new possible prostate biomarker seen at elevated levels in diseased prostate tissue, but not in healthy prostate tissue in serum are reported using paper spray ionization.

Direct detection of benzene, toluene, and ethylbenzene at trace levels in ambient air by atmospheric pressure chemical ionization using a handheld mass spectrometer

The capabilities of a portable mass spectrometer for real-time monitoring of trace levels of benzene, toluene, and ethylbenzene in air are illustrated. An atmospheric pressure interface was built to implement atmospheric pressure chemical ionization for direct analysis of gas-phase samples on a previously described miniature mass spectrometer (Gao et al. Anal. Chem.2006,78, 5994–6002). Linear dynamic ranges, limits of detection and other analytical figures of merit were evaluated: for benzene, a limit of detection of 0.2 parts-per-billion was achieved for air samples without any sample preconcentration. The corresponding limits of detection for toluene and ethylbenzene were 0.5 parts-per-billion and 0.7 parts-per-billion, respectively. These detection limits are well below the compounds’ permissible exposure levels, even in the presence of added complex mixtures of organics at levels exceeding the parts-per-million level. The linear dynamic ranges of benzene, toluene, and ethylbenzene are limited to approximately two orders of magnitude by saturation of the detection electronics.

Detection of explosives as negative ions directly from surfaces using a miniature mass spectrometer.

A miniature mass spectrometer was modified by incorporating a conversion dynode detector system and the appropriate electronics to allow the detection of negatively charged ions. The system was fitted with a discontinuous atmospheric pressure interface to allow external ionization by desorption electrospray ionization (DESI). It was used to identify the explosives 2,4,6-trinitrotoluene (TNT), 2,4,6-trinitrophenyl-N-methylnitramine (Tetryl), and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocane (HMX) present in trace amounts on surfaces (500 pg/cm(2) to 1 microg/cm(2)) both individually and as components of mixtures. Detection of explosives was demonstrated in the presence of an interfering matrix. A large surface (5 cm x15 cm) on which 1 microg/cm(2) samples of TNT, Tetryl, and HMX had been spotted randomly was interrogated in 22 s in the full scan mode, and signals characteristic of each of the explosives were observed in the DESI mass spectrum.

Induced Nanoelectrospray Ionization for Matrix‐Tolerant and High‐Throughput Mass Spectrometry

No-contact rule: the title method is ultra-sensitive, high-throughput (4 samples per second), easily multiplexed, and is compatible with serum, urine, and concentrated salt solutions. Other features of this method, which avoids physical contact between the electrode and the solvent, include sample economy and the ability to produce both positive and negative-ion spectra in one cycle.

Gas-flow assisted ion transfer for mass spectrometry.

Methods and devices that use gas flows to collect ions and transfer them over long distances for mass spectrometric analysis have been developed. Gas flows derived from the ionization source itself or provided by means of additional pumping were used to generate a laminar flow inside cylindrical tube. Hydrodynamic simulations and experimental tests demonstrate that laminar flow can transfer ions over long distance. The typical angular discrimination effects encountered when sampling ions from ambient ionization sources are minimized, and the sampling of relatively large surface areas is demonstrated with desorption electrospray ionization (DESI). Ion transfer over 6 m has been achieved and its application to multiplexed chemical analysis is demonstrated on samples at locations remote from the mass spectrometer.

Quenching the Chemiluminescence of Acridinium Ester by Graphene Oxide for Label-Free and Homogeneous DNA Detection

It was found that graphene oxide (GO) could effectively quench the chemiluminescence (CL) emission from a acridinium ester (AE)-hydrogen peroxide system. By taking advantage of this quenching effect, as a proof of concept, a label-free and homogeneous DNA assay was developed for the detection of Mycobacterium tuberculosis DNA. In the absence of target DNA, both probe DNA and AE were absorbed on the surface of GO, producing a weak CL emission owing to the CL quenching effect of GO. However, in the presence of target DNA, a double-stranded structure of DNA was generated, leading to the release of the oligonucleotide from the GO surface. AE favors binding with double-stranded DNA, which will be released from the GO surface; thus, the quenching effect of GO will be no longer effective and a strong CL signal can be observed. This assay can detect M. tuberculosis DNA with a detection limit of 0.65 nM. This sensitivity is lower than that of previously reported electrochemical detection.

Enantiomeric separation of β-blockers by HPLC using (R)-1-naphthylglycine and 3,5-dinitrobenzoic acid as chiral stationary phase

Abstract Direct liquid chromatographic separations of the enantiomers of metoprolol and bisoprolol have been developed, using (R)-1-naphthylglycine and 3,5-dinitrobenzoic acid as chiral stationary phase (CSP). The separations were achieved in a normal phase system employing a mobile phase containing n-hexane, 1,2-dichloroethane and methanol. Column efficiency was strongly dependent on the composition of the mobile phase. The eluent contents of methanol and of 1,2-dichloroethane were optimized, and so was flow-rate and column temperature. Under the optimal conditions, linear responses for (R)-metoprolol and (S)-metoprolol are obtained in the range of 0.079–1.38 and 0.015–5.80 mg/ml, with detection limits of 0.008 and 0.002 mg/ml, respectively. As for bisoprolol, the linear ranges of (R)-isomer and (S)-isomer are 0.05–1.31 and 0.02–1.00 mg/ml with detection limits of 0.001 and 0.008 mg/ml, respectively. The relative standard deviation (R.S.D.) of each enantiomer did not exceed 0.90%. The method has been successfully applied to the determination of enantiomers in pharmaceuticals.

Hand-held mass spectrometer for environmentally relevant analytes using a variety of sampling and ionization methods

A recently developed hand-held, rectilinear ion trap mass spectrometer, capable of performing in situ analysis, has been evaluated for a variety of environmentally relevant analytes. Different sampling and ionization methods were implemented, demonstrating the considerable versatility of this instrument. A discontinuous (viz. pulsed) atmospheric pressure inlet (DAPI) was used to introduce externally-generated analyte ions. Nitro compounds were ionized by electrosonic spray ionization (ESSI) yielding the protonated and sodiated forms of the molecular ion, as well as fragment ions. The amines 2,2,6,6-tetramethylpiperidine, triethylamine and 2,6-diphenylpyridine showed low parts per billion (ppb) detection limits. Vapor phase external ionization was used to examine the chemical warfare simulant dimethyl methylphosphonate and the insect repellant N,N-diethyl-m-toluamide. Membrane introduction mass spectrometry (MIMS) was used as the introduction system for hydrophobic analytes using a selectively permeable (polydimethylsiloxane) membrane placed within the vacuum manifold with subsequent ionization of the thermally desorbed neutral compounds inside the ion trap. MIMS allowed the quantitation of trace levels (a few ppb) of fluorinated compounds in the vapor phase. MIMS was also applied to the quantitation of aqueous polycyclic aromatic hydrocarbons (PAHs) with limits of detection again in the low ppb range for naphthalene, acenaphthene, anthracene and phenanthrene.

N-(Aminobutyl)-N-(ethylisoluminol) and hemin dual-functionalized graphene hybrids with high chemiluminescence.

Herein, we report a facile strategy for the synthesis of dual-functionalized graphene hybrids (A-H-GNs) based on interactions among graphene, N-(aminobutyl)-N-(ethylisoluminol) (ABEI) and hemin. The multilayers of a chemiluminescence reagent (ABEI) and a catalyst (hemin) were accumulated on the graphene surface, resulting in high chemiluminescence performance, good solubility and stability in aqueous solution.

Reactive paper spray mass spectrometry for in situ identification of quinones

RATIONALE The polycyclic aromatic hydrocarbons quinones are reported to be harmful and could cause mutations and cancer via the generation of reactive oxygen species through their redox cycle in human body. For detection by gas chromatography and high-performance liquid chromatography mass spectrometry (MS), sample pretreatments and chromatographic separation prior to MS are generally required, which makes the whole analytical process laborious and time-consuming, resulting in difficulties for fast screening targets from complicated matrices. Thus facile, rapid and reliable MS methods for detection of quinones in complicated matrices are in great demand. METHODS Reactive paper spray mass spectrometry is reported for rapid identification and quantification of quinones in complicated matrices. The method is based on an in situ derivatization reaction between cysteamine and quinones prior to analysis with paper spray mass spectrometry. With the addition of an easily charged chemical tag, the ionization efficiency of analysts is greatly improved. Due to the high ionization efficiency of the drivatives, quinones in complicated matrices could be detected rapidly without any pretreatment. RESULTS Under the optimized experimental conditions, the linear dynamic ranges for both 1,4-benzoquinone and 1,4-naphthoquinone are 0.4-40 ng and that for 1,4-anthraquinone is 0.4-20 ng. Limits of detection for these three analytes were measured to be 160, 40 and 200 pg using methyl-p-benzoquinone as internal standard. The capability to conduct MS analysis under ambient pressure is illustrated by identification of 1,4-naphthoquinone and 1,4-anthraquinone in raw urine, raw serum and cell culture medium. CONCLUSIONS Reactive paper spray could be applied to fast screening of quinones from complicated matrices. Therefore, we believe that reactive paper spray mass spectrometry might be potentially useful in the fields of environmental sciences, metabolomics and clinic analysis.