共查询到20条相似文献,搜索用时 31 毫秒
1.
Valentine NB Wahl JH Kingsley MT Wahl KL 《Rapid communications in mass spectrometry : RCM》2002,16(14):1352-1357
In this study various methods of sample preparation and matrices were investigated to determine optimum collection and analysis criteria for fungal analysis by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Intact spores and/or hyphae of Aspergillus niger, Rhizopus oryzae, Trichoderma reesei and Phanerochaete chrysosporium were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The fungal samples were applied to the MALDI sample target as untreated, sonicated, or acid/heat treated samples, or blotted directly from the fungal culture with double-stick tape. Ferulic acid or sinapinic acid matrix solution was layered over the dried samples and analyzed by MALDI-MS. Statistical analysis showed that simply using double-stick tape to collect and transfer to a MALDI sample plate typically worked as well as the other preparation methods, and required the least sample handling. 相似文献
2.
Jasmin Kemptner Martina Marchetti‐Deschmann Robert Mach Irina S. Druzhinina Christian P. Kubicek Günter Allmaier 《Rapid communications in mass spectrometry : RCM》2009,23(6):877-884
Unambiguous identification of mycotoxin‐producing fungal species as Fusarium is of great relevance to agriculture and the food‐producing industry as well as in medicine. Protein profiles of intact fungal spores, such as Penicillium, Aspergillus and Trichoderma, derived from matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) were shown to provide a rapid and straightforward method for species identification and characterization. In this study, we applied this approach to five different Fusarium spp. strains which are known to affect the growth of different grain plants. To obtain a suitable MALDI matrix system and sample preparation method, thin‐layer, dried‐droplet and sandwich methods and several MALDI matrices, namely CHCA, DHB, FA, SA and THAP dissolved in various solvent mixtures (organic solvents such as ACN, MeOH, EtOH and iPrOH and for the aqueous phase water and 0.1% TFA), were evaluated in terms of mass spectrometric pattern and signal intensities. The most significant peptide/protein profiles were obtained with 10 mg ferulic acid (FA) in 1 mL ACN/0.1% TFA (7:3, v/v) used as matrix system. Mixing the spores with the matrix solution directly on the MALDI target (dried‐droplet technique) resulted in an evenly distributed spores/matrix crystal layer, yielding highly reproducible peptide/protein profiles from the spore surfaces. Numerous abundant ions throughout the investigated m/z range (m/z 1500–15 000) could be detected. Differences in the obtained mass spectral patterns allowed the differentiation of spores of various Fusarium species. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献
3.
Yaouba A Tatsadjieu LN Dongmo PM Etoa FX Mbofung CM Zollo PH Menut C 《Natural product communications》2011,6(9):1367-1371
Investigations were conducted to determine the chemical composition, antifungal and antiradical activities of the essential oil extracted from the fresh leaves of Clausena anisata (Willd.) Hook. F. ex Benth (from Cameroon) against Aspergillus flavus, A. niger, A. parasiticus and Fusarium moniliforme. The essential oil obtained by hydrodistillation was analysed by GC and GC/MS. The disc diffusion method was used to evaluate the fungal growth inhibition at various concentrations of the oil while the antiradical activity of the essential oil was studied by the DPPH (diphenyl picryl hydrazyl) method. The main components obtained were E-ocimenone (15.1%), Z-ocimenone (11.5%), gamma-terpinene (11.4%) and germacrene D (10.9%). After 10 days of incubation on essential oil supplemented medium, the growth of A. flavus, A. niger, A. parasiticus and F. moniliforme were totally inhibited by 4, 5, 5 and 5 mg/mL of C. anisata essential oil, respectively. The antiradical activity of C. anisata essential oil (SC50 = 5.1 g/L) was less than that of butylated hydroxyl toluene (BHT), which was used as the reference compound (SC50 = 0.007 g/L). Results obtained in the present study indicate the possibility of exploiting C. anisata essential oil to fight strains of A. flavus, A. niger, A. parasiticus and F. moniliforme responsible for biodeterioration of stored food products. 相似文献
4.
Stowers MA van Wuijckhuijse AL Marijnissen JC Scarlett B van Baar BL Kientz CE 《Rapid communications in mass spectrometry : RCM》2000,14(10):829-833
Matrix-assisted laser desorption/ionization (MALDI) mass spectra were obtained from single biological aerosol particles using an aerosol time-of-flight mass spectrometer (ATOFMS). The inlet to the ATOFMS was coupled with an evaporation/condensation flow cell that allowed the aerosol to be coated with matrix material as the sampled stream entered the spectrometer. Mass spectra were generated from aerosol composed either of gramicidin-S or erythromycin, two small biological molecules, or from aerosolised spores of Bacillus subtilis var niger. Three different matrices were used: 3-nitrobenzyl alcohol, picolinic acid and sinapinic acid. A spectrum of gramicidin-S was generated from approximately 250 attomoles of material using a molar ratio of 3-nitrobenzyl alcohol to analyte of approximately 20:1. A single peak, located at 1224 Da, was obtained from the bacterial spores. The washing liquid and extract solution from the spores were analyzed using electrospray mass spectrometry and subsequent MS/MS product ion experiments. This independent analysis suggests that the measured species represents part of the B. subtilis peptidoglycan. The on-line addition of matrix allows quasi-real-time chemical analysis of individual, aerodynamically sized particles, with an overall system residence time of less than 5 seconds. These results suggest that a MALDI-ATOFMS can provide nearly real-time identification of biological aerosols. Copyright 2000 John Wiley & Sons, Ltd. 相似文献
5.
Eric B. Monroe Beth Anne Koszczuk Jenna L. Losh John C. Jurchen Jonathan V. Sweedler 《International journal of mass spectrometry》2007,260(2-3):237
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) has been used for the discovery of hundreds of novel cell to cell signaling peptides. Beyond its advantages of sensitivity and minimal sample preparation requirements, MALDI MS is attractive for biological analyses as high quality mass spectra may be obtained directly from specific locations within prepared tissue sections. However, due to the large quantity of salts present in physiological tissues, these mass spectra often contain many adducts of cationic salts such as sodium and potassium, in addition to the molecular ion [M + H]+. To reduce the presence of cation adducts in MALDI mass spectra obtained directly from tissues, we present a methodology that uses a slow condensation procedure to enable the formation of distinct regions of matrix/analyte crystals and cation (salt) crystals. Secondary ion mass spectrometric imaging suggests that the salts and MALDI matrix undergo a mutually exclusive crystallization process that results in the separation of the salts and matrix in the sample. 相似文献
6.
Dorner JW 《Journal of AOAC International》2002,85(4):911-916
A method was developed for simultaneous quantitation of Aspergillus flavus/A. parasiticus and aflatoxins in peanuts. Peanut samples were ground with an equal weight of water in a vertical cutter mixer to produce a slurry. Separate subsamples were taken for dilution-plating to determine total colony forming units (CFU)/g of A. flavus/A. parasiticus and for liquid chromatographic analysis to determine aflatoxin concentrations. Dry-grinding peanuts for homogenization of aflatoxins produced high temperatures that killed most of the A. flavus/A. parasiticus propagules. Addition of water to produce a slurry kept the temperature from rising above levels that killed the fungi. A 7 min grind time provided optimal homogenization for both the fungi and aflatoxins, so long as the temperature of the slurry did not exceed 45 degrees C. In the analysis of 60 shelled peanut samples, total aflatoxin concentrations ranged from 0 to 10,000 ng/g and total A. flavus/A. parasiticus ranged from 1.4 x 10(3) to 3.2 x 10(6) CFU/g. Regression analysis showed a significant positive correlation (p < 0.0001) between the quantities of A. flavus/A. parasiticus and aflatoxin (R2 = 0.82). 相似文献
7.
Jana Chalupová Michaela Sedlářová Michaela Helmel Pavel Řehulka Martina Marchetti‐Deschmann Günter Allmaier Marek Šebela 《Journal of mass spectrometry : JMS》2012,47(8):978-986
Fast and easy identification of fungal phytopathogens is of great importance in agriculture. In this context, matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) has emerged as a powerful tool for analyzing microorganisms. This study deals with a methodology for MALDI‐TOF MS‐based identification of downy and powdery mildews representing obligate biotrophic parasites of crop plants. Experimental approaches for the MS analyses were optimized using Bremia lactucae, cause of lettuce downy mildew, and Oidium neolycopersici, cause of tomato powdery mildew. This involved determining a suitable concentration of spores in the sample, selection of a proper MALDI matrix, looking for the optimal solvent composition, and evaluation of different sample preparation methods. Furthermore, using different MALDI target materials and surfaces (stainless steel vs polymer‐based) and applying various conditions for sample exposure to the acidic MALDI matrix system were investigated. The dried droplet method involving solvent evaporation at room temperature was found to be the most suitable for the deposition of spores and MALDI matrix on the target and the subsequent crystallization. The concentration of spore suspension was optimal between 2 and 5 × 109 spores per ml. The best peptide/protein profiles (in terms of signal‐to‐noise ratio and number of peaks) were obtained by combining ferulic and sinapinic acids as a mixed MALDI matrix. A pretreatment of the spore cell wall with hydrolases was successfully introduced prior to MS measurements to obtain more pronounced signals. Finally, a novel procedure was developed for direct mass spectra acquisition from infected plant leaves. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
8.
Anu Teearu Signe Vahur Uku Haljasorg Ivo Leito Tõiv Haljasorg Lauri Toom 《Journal of mass spectrometry : JMS》2014,49(10):970-979
2,5‐Dihydroxybenzoic acid (DHB) is one of the most widely used and studied matrix compounds in matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry. However, the influence of ageing of the DHB solution on the MALDI mass spectra has not been yet systematically studied. In this work, the possible changes occurring in the acidified acetonitrile/water solution of the MALDI matrix compound DHB during 1‐year usage period have been monitored with MALDI‐Fourier transform ion cyclotron resonance mass spectrometer (MALDI‐FT‐ICR‐MS) and attenuated total reflectance Fourier transform infrared (ATR‐FT‐IR) spectroscopy. No significant ageing products have been detected. The ability of the aged DHB solution to act as a MALDI matrix was tested with two materials widely used in art and conservation – bone glue (a proteinaceous material) and shellac resin (a resinous material) – and good results were obtained. A number of peaks in the mass spectra measured from the DHB solution were identified, which can be used for internal calibration of the mass axis. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
9.
The fungal spores of Penicillium expansum, P. chrysogenum, P. citrinum, P. digitatum, P. italicum, and P. pinophilum were characterized by using matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry (MALDI-TOFMS). These fungal spores are frequently found in grain and fruit. The mass spectra of these six species were directly obtained from the intact spores without any pretreatment. The results obtained indicate that 2,5-dihydroxybenzoic acid and sinapinic acid are suitable matrices for the analysis of Penicillium spores. Characteristic ions representing the different species were obtained with sufficiently high reproducibility that these ions can be employed to identify the different fungal species. On the basis of these characteristic ions obtained from these authentic Penicillium spores, the approach was applied to characterize the fungal species contaminating the surfaces of fruit. It was demonstrated that the fungal spores directly scratched from the surfaces of fruit contaminated by unknown fungi can be rapidly identified using MALDI-TOFMS analysis without any tedious pretreatment. 相似文献
10.
The use of collision-induced dissociation, postsource decay (CID-PSD) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry for the analysis of small organic molecules is demonstrated. Three pesticides: paraquat, diquat, and difenzoquat were chosen for this study. The matrices 2,5-dihydroxybenzoic acid (DHB), alpha-cyano-4-hydroxycinnamic acid (alpha-CHCA), and sinapinic acid (SA) were selected to investigate the effect of the matrix on the CID-PSD MALDI spectra of these molecules. Alpha-CHCA and DHB were found to be appropriate matrices for the pesticides studied. Spectra for a given pesticide obtained from different matrices were compared with each other, and the differences between them are discussed. A comparison of CID-PSD MALDI with fast-atom bombardment MS/MS spectra is presented; the agreement of pesticide fragmentation patterns between the two methods indicates that CID-PSD MALDI MS is a reliable and efficient technique for structural elucidation of small molecules. 相似文献
11.
Raju NP Mirza SP Vairamani M Ramulu AR Pardhasaradhi M 《Rapid communications in mass spectrometry : RCM》2001,15(19):1879-1884
A preliminary investigation of the use of 5-ethyl-2-mercaptothiazole as matrix in matrix-assisted laser desorption/ionization (MALDI) of a broad spectrum of analytes is reported. The analytes studied are substance P, insulin, beta-cyclodextrin, triacylglycerols of coconut oil and polypropylene glycol 2000 (PPG 2000). In the positive ion mass spectra of the matrix/analyte combinations, the formation of [M + H]+ and [M + cation]+ species were observed and compared with those obtained by using well-established matrices such as alpha-cyano-4-hydroxycinnamic acid, genticic acid, sinapinic acid and dithranol. In addition, the usefulness of this new matrix for MALDI in negative ion mode is also described using substance P and beta-cyclodextrin as examples. 相似文献
12.
Using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), this study analyzed the saliva obtained from patients with oral cancer and compared these mass spectra with those obtained from healthy controls. Saliva without pre-treatment was mixed directly with a sinapinic acid matrix. Alpha-amylase (57 kDa) dominated the high mass range in the MALDI mass spectra of the saliva from healthy subjects, but the peak was suppressed for patients with oral cancer and was replaced by a peak at m/z 66 k in the spectra of patients' samples (15 out of 20). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with in-gel tryptic digestion combined with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) was employed to characterize this 66-kDa protein, which was thus shown to be albumin. However, based on SDS-PAGE results, concentrations of both alpha-amylase and albumin in patients' saliva were significantly higher than those in healthy subjects. This discrepancy was shown to be due to MALDI suppression effects due to the albumin. MALDI-MS thus has potential as a possible rapid diagnostic screening tool for oral cancer. 相似文献
13.
Ting Wu Chuanjing Zhang Hongxin Ren Yidan Xi Yiping Du 《International Journal of Polymer Analysis and Characterization》2017,22(2):160-168
Solvent effect is one of the important factors in sample preparation which may affect matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectra of synthetic polymers. MALDI imaging, a useful imaging tool for discovering biomarkers in tissues, is applied here for better comprehension of solvent effect in polymer analysis by MALDI-TOF mass spectrometry. Nylon-6 was chosen as a model polymer for the study of solvent effect. Its MALDI mass spectra in different solvents were performed. MALDI imaging analysis was performed for studying the incorporation of analytes into matrix crystals in different solvent combinations. Specifically, the colocalization of matrix and analyte was obtained through Pearson’s correlation (PC) coefficient analysis of their MALDI images. The results demonstrated that satisfactory spectra were obtained in higher PC value conditions. PC decreased along with an increase in the ratio of poor solvent, which suggested that we should minimize the poor solvent ratio to obtain better MALDI spectra. 相似文献
14.
It has been described that ion yield in both positive- and negative-ion matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) of peptides is often inhibited by trace amounts of alkali metals and that the MALDI mass spectra are contaminated by the interfering peaks originating from traces of alkali metals, even when sample preparation is carefully performed. Addition of serine to the commonly used MALDI matrix alpha-cyano-4-hydroxycinnamic acid (CHCA) significantly improved and enhanced the signals of both protonated and deprotonated peptides, [M+H](+) and [M-H](-). The addition of serine to CHCA matrix eliminated the alkali-metal ion adducts, [M+Na](+) and [M+K](+), and the CHCA cluster ions from the mass spectra. Serine and serinephosphate as additives to CHCA enhanced and improved the formation of molecular-related ions of phosphopeptides in negative-ion MALDI mass spectra. 相似文献
15.
Solid phase microextraction with matrix assisted laser desorption/ionization introduction to mass spectrometry and ion mobility spectrometry 总被引:1,自引:0,他引:1
Solid phase microextraction (SPME) with matrix assisted laser desorption/ionization (MALDI) introduction was coupled to mass spectrometry and ion mobility spectrometry. Nicotine and myoglobin in matrix 2,5-dihydroxybenzonic acid (DHB), enkephalin and substance P in alpha-cyano-4-hydroxy cinnaminic acid were investigated as the target compounds. The tip of an optical fiber was silanized for extraction of the analytes of interest from solution. The optical fiber thus served as the sample extraction surface, the support for the sample plus matrix, and the optical pipe to transfer the laser energy from the laser to the sample. The MALDI worked under atmospheric pressure, and both an ion mobility spectrometer and a quadrupole/time-of-flight mass spectrometer were used for the detection of the SPME/MALDI signal. The spectra obtained demonstrate the feasibility of the SPME with MALDI introduction to mass spectrometry instrumentation. 相似文献
16.
Experiments have been performed to elucidate the mechanism of proton transfer in ternary clusters containing the matrix-assisted laser-desorption ionization (MALDI) matrix sinapinic acid, nonchromophoric analytes (proline, methionine, and prolylmethionine), and argon. To investigate the mechanism of intracluster proton transfer, ionizing laser power studies were performed at 266 and 355 nm. Baseline studies show that two photons are required at both wavelengths for the formation of sinapinic acid radical cations from sinapinic acid/argon clusters. Studies of the ternary sinapinic acid/biomolecule/argon clusters show that, in all cases, the photon dependence for protonation of the biomolecule is the same as that for formation of the sinapinic acid radical cation. Furthermore, the slopes of the power plots are generally between 1.5 and 2.0, consistent with a two photon ionization process. No evidence of negative ion formation is detected in the negative ion mass spectra. The combined results are consistent with a mechanism of biomolecular intracluster protonation via proton transfer from the photoionized sinapinic acid radical cation. Wavelength dependent trends in matrix and analyte fragment ion formation in conventional MALDI mass spectra and the cluster proton transfer mass spectra were noted. The possible contribution of cluster proton transfer to the analyte protonation mechanism in conventional MALDI is discussed. 相似文献
17.
金黄色葡萄球菌及其甲氧苯青霉素耐药性的MALDI-TOF MS鉴定 总被引:7,自引:1,他引:7
用MALDI-TOS MS细菌指纹图谱鉴定细菌,建立区分金黄色葡萄球菌甲氧苯青霉素耐药株和敏感株的MALDI-TOF MS分析方法,检测了76株从临床标本中分离得到的金黄色葡萄球菌,用软件进行聚类分析,以nuc(耐热核酸酶)基因和mecA(耐药)基因聚合酶链反应(PCR)检测结果为参照,74%的菌株经MALDI-TOF MS给出了正确的鉴定结果;金黄色葡萄球菌甲氧苯青霉素耐药株和敏感株的质谱图有很大差别,各自有其特征峰;经过软件聚类分析,76株实验菌株划敏感群和耐药群;与PCR检测结果对照,有7株菌PCR检测mecA基因为阴性,而经MALDI-TOF MS鉴定为耐药株,表型鉴定表明其中有5株为敏感株;利用细菌指纹图谱和数据库检索对大多数菌株实现了正确鉴定;MALDI-TOF MS分辨率高,甚至可以区分株间的差异,实现了区分金黄色葡萄球菌甲氧苯青霉素耐药株和敏感株;结果表明MALDI-TOF MS提供了一个很有前景的鉴定细菌的快速方法。 相似文献
18.
Heike Brandt Thomas Ehmann Matthias Otto 《Rapid communications in mass spectrometry : RCM》2010,24(16):2439-2444
The principle relating to the selection of a proper matrix, cationization reagent, and solvent for matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) of synthetic polymers is still a topic of research. In this work we focused on the selection of a suitable MALDI solvent. Polystyrene PS7600 and poly(ethylene glycol) PEG4820 were analyzed by MALDI‐TOF MS using various solvents which were selected based on the Hansen solubility parameter system. For polystyrene (PS), dithranol was used as the matrix and silver trifluoroacetate as the cationization reagent whereas, for poly(ethylene glycol) (PEG), the combination of 2,5‐dihydroxybenzoic acid and sodium trifluoroacetate was used for all experiments. When employing solvents which dissolve PS and PEG, reliable MALDI mass spectra were obtained while samples in non‐solvents (solvents which are not able to dissolve the polymer) failed to provide spectra. It seems that the solubility of the matrix and the cationization reagent are less important than the polymer solubility. Copyright © 2010 John Wiley & Sons, Ltd. 相似文献
19.
Russell WK Russell DH Busby MB Kolberg A Li S Maynard DK Sanchez-Vindas S Zhu W Vigh G 《Journal of chromatography. A》2001,914(1-2):325-330
Artifact-free, high-resolution matrix-assisted laser desorption ionization (MALDI) time-of-flight mass spectra have been obtained for the labile, single-isomer, tert.-butyldimethylsilyl ether derivatives of alpha-, beta- and gamma-cyclodextrins by optimizing the MALDI sample preparation method. 2,5-Dihydroxybenzoic acid, a 3:1 mixture of 2,5-dihydroxybenzoic acid and 1-hydroxyisoquinoline, and 2,4,6-trihydroxyacetophenone were investigated as MALDI matrices with methanol and acetonitrile as matrix solvents. Partial-to-complete loss of the tert.-butyldimethylsilyl groups was observed when the commonly used 2,5-dihydroxybenzoic acid was the MALDI matrix and/or methanol was the solvent, both with and without trifluoroacetic acid as additive. Loss of the labile tert.-butyldimethylsilyl groups was avoided with 2,4,6-trihydroxyacetophenone as MALDI matrix and acetonitrile as matrix solvent. Good ion intensities were achieved for the (M+Na)+ and (M+K)+ quasimolecular ions in the positive-ion mode. Minor byproducts were observed in some of the samples and the information was used to aid the optimization of the synthetic work. 相似文献
20.
Hongjuan Dong Jasmin Kemptner Martina Marchetti-Deschmann Christian Peter Kubicek Günter Allmaier 《Analytical and bioanalytical chemistry》2009,395(5):1373-1383
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI–TOF MS) has been proved to be a powerful
tool for the identification and characterization of microorganisms based on their surface peptide/protein pattern. Because
of the complexity of microorganisms, there are no standardized protocols to acquire reproducible peptide/protein profiles
for a broad range of microorganisms and for fungi in particular. Small variations during MALDI MS sample preparation affect
the quality of mass spectra quite often. In this study, we were aiming to develop a sample preparation method for the analysis
of colored, a quite often observed phenomenon, and mycotoxin-producing Fusarium conidia spores using MALDI–TOF MS. Different washing solvent systems for light- and deep-colored (from slightly orange to
red-brown) conidia spores and connected sample deposition techniques were evaluated based on MS reproducibility and number
and intensities of peaks. As a method of choice for generation of reproducible and characteristic MALDI–TOF mass spectra,
the use of a washing process for colored Fusarium conidia spores with acetonitrile/0.5% formic acid (7/3) was found and subsequently combined with two-layer volume technique
(spores/matrix (ferulic acid) solution was deposited onto a MALDI target, and after solvent evaporation, a second matrix layer
was deposited). With the application of this sample preparation method, for deep-colored Fusarium species, 19 abundant molecular ions in the m/z range 2,000–10,000 were always detected with an S/N ratio of 3:1 or better. Finally this optimized sample preparation for
the first time provided mass spectrometric fingerprints of strongly colored Fusarium conidia spores resulting in the possibility of differentiation of such spores at the species level.
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