Eleonora Mazzucco

Signal suppression/enhancement in high-performance liquid chromatography tandem mass spectrometry.

The review discusses the pitfalls of the matrix effect in mass spectrometry detection hyphenated to liquid chromatography separation. Matrix effect heavily influences both qualitative and quantitative analyses, giving rise to suppression or enhancement of the signal. As generally recognised, the predominant cause is the presence of undesired components that co-elute in the chromatographic separation and alter the ionisation process. The interfering species can be components of the sample, compounds released during the pre-treatment/extraction process or reagents added to the mobile phase to improve chromatographic resolution. The different mechanisms proposed in literature to explain the suppression or the enhancement of the signal both in electrospray and atmospheric pressure chemical ionisations are presented and the results observed in the different experimental conditions are compared and discussed. All data together lead to conclude that the chemical properties of the target analyte, the kind of matrix, the matrix to analyte concentration ratio, the extraction process, the chromatographic conditions as well as the kind of the mass spectrometry instrumentation and the ionisation conditions can play a role. Likely all these potential causes act in a synergic way and the final effect observed is hardly due to only one of them. Depending on an unpredictable combination of conditions, signal suppression or enhancement can be observed. The review discusses the matrix effects observed in HPLC-MS and HPLC-MS/MS analysis proposes hypotheses to explain the observed behaviours and proposes methods and strategies to overcome the matrix effects.

Determination of perfluorochemicals in biological, environmental and food samples by an automated on-line solid phase extraction ultra high performance liquid chromatography tandem mass spectrometry method

A rapid on-line solid phase extraction ultra high performance liquid chromatography tandem mass spectrometry method was developed for the identification and quantitation of nine perfluorinated compounds in matrices of environmental, biological and food interest. Pre-treatment, solid phase extraction, chromatographic and mass detection conditions were optimised, in order to apply the whole methodology to the analysis of different matrices. Particular attention was devoted to the evaluation of matrix effect and the correlated phenomena of ion enhancement or suppression in mass spectrometry detection. LOD and LOQ range from 3 to 15ngL(-1) and from 10 to 50ngL(-1), respectively. Method detection limits (MDLs) were also calculated for each kind of matrix. The recovery, evaluated for each analyte, does not depend on analyte concentration in the explored concentration range: average R¯% values are always greater than 82.9%. In the whole, the results obtained for samples of river waters, blood serum, blood plasma, and fish confirm the ubiquitous presence of perfluorinated compounds, as recently denounced by many sources.

A new hydrophilic interaction liquid chromatography tandem mass spectrometry method for the simultaneous determination of seven biogenic amines in cheese

An HILIC-PI APCI MS/MS method is developed for the determination of seven biogenic amines (cadaverine, histamine, putrescine, spermidine, spermine, tryptamine and tyramine) in cheese. Their presence and relative amounts give useful information about freshness, level of maturing, quality of storage and cheese typicization. The major drawback in the analysis is represented by the relevant matrix effect and the general unbalanced concentrations of the different amines in cheese. The method proposed represents an improvement with respect to an HPLC-MS/MS method already developed in this laboratory. The new method permits better sensitivities it makes use of a Waters Atlantis HILIC (150.0 mm x 2.1 mm i.d., 3 microm) stationary phase and of a mobile phase of acetonitrile and ammonium formate 50.0 mM in ultrapure water brought to pH 4.00 for formic acid, flowing under gradient conditions. The chromatographic system is interfaced with a 3200QTrap LC-MS/MS system (Applied Biosystem, Foster City, CA, USA) by a Turbo V interface equipped with Heated Nebuliser (APCI) and Turbo Ion Spray (TIS) probes. LOQ values lower than 10 microg L(-1) are obtained. The method is applied in the analysis of Castelmagno cheese.

Automated Online Solid Phase Extraction Ultra High Performance Liquid Chromatography Method Coupled with Tandem Mass Spectrometry for Determination of Forty-Two Therapeutic Drugs and Drugs of Abuse in Human Urine

The study deals with a fast (analysis times of around 11 min) simultaneous identification and quantification in human urine of 42 drugs (21 therapeutic and 21 of abuse), through an automated online solid phase extraction ultra high performance liquid chromatography method coupled with tandem mass spectrometry (SPE UHPLC-MS/MS). In the method validation, particular attention was devoted to the matrix effect, through matrix-matched calibration in blank urine, suitably diluted. For all the abuse drugs investigated, the limit of quantitation (LOQ) values are lower than the legal threshold concentration levels, making the method suitable for routine control. The whole procedure was applied in the analysis of urine of patients positive to the I level screening test.

High-Performance Liquid Chromatography−Ultraviolet Detection Method for the Simultaneous Determination of Typical Biogenic Amines and Precursor Amino Acids. Applications in Food Chemistry

A reversed-phase high-performance liquid chromatography (HPLC) method was developed for the simultaneous determination in food of biogenic amines and their precursor amino acids after a precolumn derivatization with dansyl chloride. The chromatographic conditions, selected to be suitable for mass spectrometry detection, were optimized through experimental design and artificial neural networks. The HPLC-UV method was validated by comparing the separation results with those obtained through a HPLC method, working under the same chromatographic conditions but employing mass spectrometry detection. The HPLC-UV method was then applied to the analysis of different food samples, namely, cheese, clams, salami, and beer. For all of the matrices, recoveries (relative standard deviation always <5%) always >92% were obtained. The results are discussed as a function of the total biogenic amine content and of the concentration ratio between amines and precursor amino acids.

Simultaneous determination of thirteen polycyclic aromatic hydrocarbons and twelve aldehydes in cooked food by an automated on-line solid phase extraction ultra high performance liquid chromatography tandem mass spectrometry.

An on-line solid phase extraction (SPE) ultra high performance liquid chromatography tandem mass spectrometry method has been developed for the simultaneous identification and determination of thirteen polycyclic aromatic hydrocarbons (PAHs) and twelve aldehydes (derivatized with 2,4-dinitrophenylhydrazine). The chromatographic conditions have been optimized to obtain the maximum of sensitivity and resolution taking into account the different retention interactions and the different ionization conditions of PAHs and derivatized aldehydes. LOD values ranging from 0.028 to 0.768 μg L(-1) for PAHs and from 0.002 to 0.125 μg L(-1) for aldehydes were obtained. The resolution permitted the separation of four couples of PAH isomers. Sample pre-treatment and SPE were optimized in order to apply the whole methodology to the analysis of different food matrices as salmon, frankfurter, steak, and pork chop, subjected to different cooking modes (smoked, grilled, cooked in oil or in butter). Particular attention was devoted to the evaluation of matrix effect that was significantly reduced through the on-line SPE treatment. For each food matrix the method detection limits, the method quantitation limits, and the recovery R were evaluated. R was shown not to depend on analyte concentration in the explored concentration range (LOQ - 50.000 μg L(-1)): the average R percent ranges from 70.6% to 120.0%.

Sun light degradation of 4-chloroaniline in waters and its effect on toxicity. A high performance liquid chromatography ― Diode array ― Tandem mass spectrometry study

This paper studies the degradation reactions that 4-chloroaniline can naturally undergo in waters for the action of sun light. 10.00 mg L(-1) 4-chloroaniline aqueous solution, without any addition of organic solvent, are undergone to photoirradiation under conditions that simulate sun light. The degradation pathway, followed by HPLC-DAD-MS/MS methods, is complex since the pollutant gives rise to many photoproducts: the predominant species are characterized by m/z values of 217 (P5) and 218 (P6) and are compatible with dimeric structures of 4-chloroaniline. Vibrio fischeri tests indicate that the photoproducts of 4-chloroaniline are characterized by a toxicity level significantly greater than the precursor.

Hydrolytic and Photoinduced Degradation of Tribenuron Methyl Studied by HPLC-DAD-MS/MS

The paper studies, with the help of HPLC-DAD-MS/MS technique, the hydrolytic and photoinduced degradation processes that take place in aqueous solutions of tribenuron methyl, both when preserved in the dark and when undergoing solar box irradiation under conditions that simulate sun light. The results indicate that the degradation products formed by hydrolysis alone and by photoirradiation are the same, but kinetics of the hydrolysis reaction is much slower. The degradation products are identified as 2-methoxy-4-methylamino-6-methyl-1,3,5-triazine (P1), methyl 2-aminosulfonylbenzoate (P2), and saccharin (P3) and quantified. Ecotoxicological biotests performed on 0.1 µg L−1 photoirradiated solutions of the herbicide give a border line toxicity situation comparable to that of the precursor and indicate that the herbicide is characterized by low persistence in the environment, as required. Its degradation, however, does not lead to mineralization but to the formation of products of comparable toxicity. To evaluate the matrix effects, the photodegradation of the herbicide is also studied in the presence of rice paddy waters: the process is slower than in ultrapure water but leads to the same products. Experiments performed for comparison by irradiating ultrapure water solutions with UV lamp (254 nm) show that the degradation process is not only faster with respect to sunlight, but gives a different pathway, without in anyway leading to mineralization.

A new on-line solid phase extraction high performance liquid chromatography tandem mass spectrometry method to study the sun light photodegradation of mono-chloroanilines in river water.

The study investigates the natural photodegradation pathway of mono-chloroanilines in river waters, with the aim to identify the predominant photoproducts formed. At this purpose a new sensitive on-line SPE HPLC-MS/MS method has been developed with LOQ values equal or lower than the legal threshold concentration levels allowed for mono-chloroanilines in waters. The degradation processes of o-, m- and p-chloroaniline have been investigated subjecting their solutions, prepared both in ultrapure and in river water, to sun light irradiation simulated by a solar box system. The SPE HPLC-MS/MS methodology allowed to evaluate the degradation kinetics, to identify the predominant photodegradation products and to propose the chemical structures. Two photoproducts (aniline and 3-aminophenol), for which standards are available, have also been quantified.

Development of a technique based on multi-spectral imaging for monitoring the conservation of cultural heritage objects

A new approach for monitoring the state of conservation of cultural heritage objects surfaces is being developed. The technique utilizes multi-spectral imaging, multivariate analysis and statistical process control theory for the automatic detection of a possible deterioration process, its localization and identification, and the wavelengths most sensitive to detecting this before the human eye can detect the damage or potential degradation changes occur. A series of virtual degradation analyses were performed on images of parchment in order to test the proposed algorithm in controlled conditions. The spectral image of a Dead Sea Scroll (DSS) parchment, IAA (Israel Antiquities Authority) inventory plate # 279, 4Q501 Apocryphal Lamentations B, taken during the 2008 Pilot of the DSS Digitization Project, was chosen for the simulation.