Evaluation of Two Fully Automated Setups for Mycotoxin Analysis Based on Online Extraction-Liquid Chromatography-Tandem Mass Spectrometry.

Mycotoxins are secondary metabolites of fungi species widely known for their potentially toxic effects on human health. Considering their frequent presence in crops and their processed food, monitoring them on food-based matrices is now an important topic. Within such a context, the sample preparation step is usually mandatory before the chromatographic analysis, due to the complexity of matrices such as nuts, cereals, beverages, and others. For these reasons, we herein present the evaluation of two greener setups, based on the automation and miniaturization of the sample preparation step for mycotoxin analysis in different beverages. Firstly, we describe an analytical method based on a multidimensional assembly, coupling a lab-made microextraction column (508 µm i.d. × 100 mm) to a UPLC-MS/MS for the analysis of ochratoxin A in beverages. This configuration used a synthesized sorbent phase containing C18-functionalized graphene-silica particles, which exhibited excellent extraction performance, as well as being reusable and cheaper than commercially available extractive phases. Sequentially, a second setup, based on a multidimensional capillary LC coupled to MS/MS, was assessed for the same purpose. In this case, a graphene oxide-based capillary extraction column (254 µm i.d. × 200 mm) was used as the first dimension, while a C18 analytical capillary column performed the mycotoxin separation in beverages. Although this second one has similarities with the first, we focused mainly on the benefits related to the link between a miniaturized/automated sample preparation device with a capillary LC-MS/MS system, which made our analysis greener. Additionally, the chromatographic efficiency could even be enhanced.

Miniaturized liquid chromatography focusing on analytical columns and mass spectrometry: A review.

The technological advances achieved over the last decades boosted the development of suitable benchtop platforms to work at miniaturized liquid chromatography scale (capillary and nano-LC). Under the right conditions, miniaturized LC can offer higher analysis efficiency resulting in superior chromatographic resolution and overall sensitivity than conventional LC. Among the main advantages are the reduced reagents and sample requirement, the decreasing on analytical column dimensions, and consequently flow rates and the easer coupling to mass spectrometry. This review describes fundamental aspects and advances over miniaturized LC technology with a focus on the last decade. Therefore, relevant characteristics of the most common analytical column, covering both filled (packed and monolithic) and open tubular (PLOT and WCOT) columns, are herein discussed. Alternatively, other modern approaches based on microchip separations or 2D configurations aiming for the sample preparation on the first dimension, are also introduced. Likewise, some positive and negative aspects of these systems over HPLC are underscored. Besides, considering the necessity to developed components to work at capillary or nanoscale, without significant dead-volumes, the most critical features of specially designed instrumentation for benchtop instruments are briefly discussed highlighting connectors, pumping, injections, oven and detection systems. Also, a more detailed section is presented focused on mass spectrometry efforts towards its miniaturization and how this trend can be useful working together with miniaturized LC. Finally, applications of capillary and nano-LC involving bioanalytical, environmental, and food methods are discussed to support the miniaturized LC as a powerful and emergent separation technique for the years ahead.

Recent advances and trends in miniaturized sample preparation techniques.

Advances in the area of sample preparation are significant and have been growing significantly in recent years. This initial step of the analysis is essential and must be carried out properly, consisting of a complicated procedure with multiple stages. Consequently, it corresponds to a potential source of errors and will determine, at the end of the process, either a satisfactory result or a fail. One of the advances in this field includes the miniaturization of extraction techniques based on the conventional sample preparation procedures such as liquid-liquid extraction and solid-phase extraction. These modern techniques have gained prominence in the face of traditional methods since they minimize the consumption of organic solvents and the sample volume. As another feature, it is possible to reuse the sorbents, and its coupling to chromatographic systems might be automated. The review will emphasize the main techniques based on liquid-phase microextraction, as well as those based upon the use of sorbents. The first group includes currently popular techniques such as single drop microextraction, hollow fiber liquid-phase microextraction, and dispersive liquid-liquid microextraction. In the second group, solid-phase microextraction techniques such as in-tube solid-phase microextraction, stir bar sorptive extraction, dispersive solid-phase extraction, dispersive micro solid-phase microextraction, and microextraction by packed sorbent are highlighted. These approaches, in common, aim the determination of analytes at low concentrations in complex matrices. This article describes some characteristics, recent advances, and trends on miniaturized sample preparation techniques, as well as their current applications in food, environmental, and bioanalysis fields.

Graphene particles supported on silica as sorbent for residue analysis of tetracyclines in milk employing microextraction by packed sorbent.

This paper describes the use of graphene-based sorbents for determination of four tetracyclines in milk. The synthesized materials were combined with microextraction by packed sorbent (MEPS) to act as the sample preparation step. The extraction performance of these sorbents was compared to commercial phases, and graphene supported on silica provided the best results. The analytical method optimization was carried out by employing experimental design. Firstly, an evaluation of the experimental variables (elution solvent, use of EDTA, ionic force, and pH of the washing solution) was made by a 24-1 factorial experimental design. The variables sampling, washing and elution cycles of MEPS were further optimized under a full 23 experimental design. The validation parameters were determined under optimized conditions resulting in a linearity ranging from 15 to 110 µg/L with R2 values above 0.98, and LOQs ranging from 0.05 to 0.9 µg/L. The accuracy ranged from 87.9 to 118.4% and intra/inter-day precision reported by the RSDs were lower than 19%. The proposed and validated method was successfully applied to the analysis of 11 milk samples from different animals, revealing traces of tetracyclines in only two of them. This study focused on the evaluation of graphene-based sorbents combined with MEPS for tetracycline analysis provided equivalent or even better results than other proposed methods, suggesting being a sensitive, fast and reliable alternative method for the determination of tetracyclines in milk samples.