A comparative study of UniSpray and electrospray sources for the ionization of neuropeptides in liquid chromatography tandem mass spectrometry.

Despite the extensive use of electrospray ionization (ESI) for the quantification of neuropeptides by liquid chromatography-tandem mass spectrometry (LC-MS/MS), poor ionization and transmission efficiency are described for this ionization interface. A new atmospheric pressure ionization source, named UniSpray, was recently developed and commercialized. In this study, the LC-MS performance of this new ionization interface is evaluated and compared with ESI for the quantification of seven neuropeptides. Besides comparison of signal intensities and charge state distributions, also signal-to-noise (S/N) ratios and accuracy and precision were assessed. Additionally, matrix effects of human precipitated plasma and rat microdialysate were evaluated as well as the effect of three supercharging agents on the ionization of the seven neuropeptides. UniSpray ionization resulted in signal intensities four to eight times higher at the optimal capillary/impactor voltage for all seven neuropeptides. S/N values at the other hand only increased by not more than a twofold when the UniSpray source was used. Moreover, UniSpray ionization resulted in a shift towards lower charge states for some neuropeptides. Evaluation of the matrix effects by a post-column infusion set-up resulted in different infusion profiles between ESI and UniSpray. The charge state distributions of the neuropeptides obtained with UniSpray are highly comparable with ESI. Finally, the effect of the supercharging agents on the ionization of the neuropeptides tends to be peptide-dependent with both ionization sources.

Comparison of electrospray and UniSpray, a novel atmospheric pressure ionization interface, for LC-MS/MS analysis of 81 pesticide residues in food and water matrices.

In mass spectrometry, the type and design of ionization source play a key role on the performance of a given instrument. Therefore, it is of paramount importance to evaluate newly developed sources for their suitability to analyze food contaminants like pesticide residues. Here, we carried out a head-to-head comparison of key extraction and analytical performance parameters of an electrospray ionization (ESI) source with a new atmospheric pressure ionization source, UniSpray (US). The two interfaces were evaluated in three matrices of different properties (coffee, apple, and water) to determine if multiresidue analysis of 81 pesticides by QuEChERS extraction and LC-MS/MS analysis could be improved. Depending on the matrix and irrespective of the chemical class, US provided a tremendous gain in signal intensity (22- to 32-fold in peak area, 6- to 7-fold in peak height), a threefold to fourfold increase in signal-to-noise ratio, a mild gain in the range of compounds that can be quantified, and up to twofold improvement of recovery. UniSpray offered comparable linearity and precision of the analyses with ESI, and did not affect the ion ratio. A gain in sensitivity of many compounds was observed with US, but in general, the two ionization interfaces did not show significant difference in LOD and LOQ. UniSpray suffered less signal suppression; the matrix effect was in average 3 to 4 times more pronounced, but showed better values than ESI. With no effect on recovery efficiency, US improved the overall process efficiency 3 to 4 times more than ESI. Graphical abstract.

Identification of the major selenium compound, Se-Methionine, in three yeast (Saccharomyces cerevisiae) dietary supplements by on-line narrowbore liquid chromatography-electrospray tandem mass spectrometry.

On-line monitoring of six Se-compounds was accomplished by using an XTerra MS C18 column coupled to electrospray tandem mass spectrometry (ES-MS-MS). In view of the nature of the compounds, the positively charged ion pairing agent tetraethylammoniumchloride (TEACl) was added to the mobile phase. The HPLC-ES-MS-MS method was optimized with six commercially available Se-compounds. Substitution of the analytical column by the narrowbore type significantly enhanced the sensitivity of the method. We were able to detect the m/z of these six molecules on-line. Furthermore, all product ions could be monitored. The method was applied to three different yeast-based supplements. They were submitted to proteolytic digestion and screened for their Se-content by HPLC-HG-AFS (hydride generation-atomic fluorescence spectrometry). By application of on-line narrowbore HPLC-electrospray tandem mass spectrometry, the main compound present in these three supplements, Se-Methionine, could be measured on its m/z and its product ions. The method can be further extended for on-line measurement of different Se-species in complex matrices

In vivo distribution and fractionation of indium in rats after subcutaneous and oral administration of [(114m)In]InAs.

Two in vivo experiments were carried out in this study. In the first experiment five rats were given two subcutaneous injections of [(114m)In]InAs. Major sites of accumulation were spleen, liver and kidney. The intracellular distribution of indium was examined by differential centrifugation. The cytoplasmic fraction contained most of the indium activity followed by the mitochondrial fraction. Both outcomes are in close agreement with the results obtained in previous studies. Chromatographic separations on a preparative size exclusion column were carried out. It was shown that indium was mostly bound to high molecular mass compounds in serum and in the cytoplasmic fraction of spleen, liver and kidney. In a second experiment five rats were given four oral doses of [(114m)In]InAs over a short period. Prior to this experiment the in vitro solubility of cold InAs in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) was determined using graphite furnace atomic absorption spectroscopy. In the case of the SGF only 1.3% of an InAs suspension dissolved after 48 hours incubation at 37 degrees C. InAs was not soluble in SIF. Uptake of InAs after oral administration was minimal (<1%). Due to incomplete removal of traces of [(114m)In]InAs from the gastrointestinal tract, it was impossible to calculate accurately the in vivo distribution over the different organs. As the uptake and consequently the activity in the organs were very low, no further chromatographic separations could be carried out. Considering this very low uptake, it can be concluded that InAs will not accumulate in the body after oral exposure.

Safety evaluation of organoarsenical species in edible Porphyra from the China Sea.

A study was carried out to determine arsenic species in Porphyra seaweed originating from the China Sea. Information about arsenic species in Porphyra was provided by HPLC-ICP-MS and ES-MS-MS. The total arsenic concentrations of Porphyra samples from five different producing areas ranged from 2.1 to 21.6 mg/kg. The analysis report also showed that arsenosugars were the only arsenic species that could be detected in all of the extracts of samples. Arsenosugar PO(4) was the major compound in most samples (0.3-13.9 mg/kg of dry weight), followed by arsenosugar OH (0.7-6.2 mg/kg of dry weight). A further experiment was done to investigate the stability of arsenosugars in the process of being heated. It was observed that the arsenosugars were stable during a short-term heating at 100 degrees C. Their stability in human ingestion was also studied. A substantial increase of dimethylarsinic acid (DMA) was detected in urine samples collected from six volunteers after the consumption of this seaweed. The results obtained indicated that arsenosugars had been metabolized to DMA, which is more toxic than arsenosugars. From this point of view, consumers should consider the possible adverse effects of edible Porphyra on human health and choose those Porphyra having lower arsenic concentrations.

A survey of arsenic species in Chinese seafood.

In the present report, thirty different types of Chinese edible seafood, including brown algae, red algae, fish, crab, shrimp, mussels, oysters, and clams, which are very popular foodstuffs in the Chinese kitchen, were examined for their total content of As as well as its different species. Total arsenic concentration in algae samples was 1.7-38.7 microg/g (dry weight), and 0.086-7.54 microg/g in fish and shellfish (wet weight), respectively. The arsenic species in seafood extracts were determined by using anion and cation exchange high performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICPMS). Arsenosugars were detected in all of the extracted algae samples (1.5-33.8 microg/g dry weight) and fish samples (0.018-0.78 microg/g wet weight). Arsenobetaine was detected in all of the extracted fish and shellfish samples (0.025-6.604 microg/g wet weight). In contrast, inorganic arsenic in fish and shellfish samples occurred at levels below 2% of the total arsenic. No inorganic arsenic was detected in the algae samples. This study provides information about the distribution pattern of arsenic species in seafood products. Since the major share of arsenic components in seafood is organic arsenic with a low toxicity, we can conclude that arsenic in seafood does not pose any risk to human health.

Urinary excretion of the uraemic toxin p-cresol in the rat: contribution of glucuronidation to its metabolization.

BACKGROUND: Increasing evidence indicates that lipophilic and/or protein-bound substances such as p-cresol are responsible for adverse physiological alterations in uraemic patients. To better understand the evolution of p-cresol disposition in renal failure and dialysis patients, it is necessary to determine its kinetic characteristics and biotransformation pathways. METHODS: We studied the biotransformation of p-cresol after intravenous injection of the compound in eight rats with normal renal function. Urine was collected in four 1 h intervals. To evaluate the presence of p-cresol metabolites, beta-glucuronidase was added to urine samples and the isolated unidentified chromatographic peak observed in previous experiments was submitted to tandem mass spectrometry (MS/MS) analysis. RESULTS: Administration of p-cresol produced a p-cresol peak and an unknown peak, suggesting biotransformation of the compound. Addition of beta-glucuronidase to urine samples and incubation at 37 degrees C resulted in a marked decrease in the unidentified peak height (P<0.001) together with an increase in p-cresol peak height (P<0.001), suggesting that the unidentified peak was composed, at least in part, of p-cresylglucuronide. Mass spectrometry (MS) and MS/MS analysis of the isolated unidentified peak confirmed the presence of p-cresylglucuronide. Linear regression between the peak height of p-cresylglucuronide before enzyme treatment and the increase in p-cresol peak height after enzyme treatment in samples incubated with beta-glucuronidase allowed us to calculate the amount of p-cresylglucuronide as its p-cresol equivalents. This revealed that 64% of the injected p-cresol was excreted as glucuronide. There was no change in peak heights when sulphatase was added to the urine. When p-cresol and p-cresylglucuronide levels were combined, approximately 85% of all administered p-cresol was recovered in the urine. In addition, the combined urinary excretion of p-cresol and p-cresylglucuronide was more than four times greater than excretion of p-cresol by itself (P<0.01). CONCLUSIONS: In rats with normal renal function, intravenous administration of p-cresol results in immediate and extensive metabolization of the compound into p-cresylglucuronide. The elimination of p-cresol from the body depends largely on the urinary excretion of this metabolite.

Fractionation of vanadium complexes in serum, packed cells and tissues of Wistar rats by means of gel filtration and anion-exchange chromatography.

Male Wistar rats were intraperitoneally injected with [(48)V]vanadium tracer to (1) investigate the distribution of vanadium over different tissues and (2) study the distribution of vanadium over the proteins and peptides in serum, packed cells and homogenates of tissues by means of liquid chromatography experiments (size exclusion, ion exchange). Target organs were primarily kidney, bone, spleen and liver. In serum we found that vanadium was mainly bound to transferrin; however, a small amount was also bound to albumin. Besides these two complexes, a significant part of vanadium occurred as readily exchangeable ("free") vanadium. In packed cells, vanadium is mainly bound to hemoglobin and to two abundant low molecular mass complexes. The chromatograms of tissues (kidney, liver, testes, spleen and lung) show similar high molecular mass complexes (vanadium co-elutes with ferritin, transferrin and hemoglobin). Between the low molecular mass complexes there are similar peaks for spleen, testes and kidneys on the one hand, and liver and lung on the other hand, albeit the differences are small. In the case of lung, there is an additional low molecular mass peak.

Arsenic speciation in chinese seaweeds using HPLC-ICP-MS and HPLC-ES-MS.

Three common Chinese edible seaweeds, one brown (Laminaria japonica) and two red (Porphyra crispata and Eucheuma denticulatum), were examined for their total arsenic content. The As species were extracted with yields of 76.4, 69.8 and 25.0%, respectively. Anion-exchange and cation-exchange high-performance liquid chromatography (HPLC) in combination with inductively coupled plasma mass spectrometry (ICP-MS) were used for the separation of the different arsenic species in two of the three seaweed extracts (Laminaria and Porphyra). The main arsenic species in the algal extracts are arseno sugars, although it has been shown that the Laminaria seaweed contains significant amounts of dimethylarsinic acid (DMA). HPLC was coupled with electrospray mass spectrometry (ES-MS) for structural confirmation of the arsenic species. The mass spectrometer settings for the arseno sugars were optimised using standards. The conclusions drawn on the basis of HPLC-ICP-MS were confirmed by the HPLC-ES-MS data. The HPLC-ES-MS method is capable of determining both arseno sugars and DMA in the seaweeds. The unknown compounds seen in the HPLC-ICP-MS chromatogram of Laminaria could not be ascribed to trimethylarsenic oxide or tetramethylarsonium ion.