Rapid Identification and Analysis of Ochratoxin-A in Food and Agricultural Soil Samples Using a Novel Semi-Automated In-Syringe Based Fast Mycotoxin Extraction (FaMEx) Technique Coupled with UHPLC-MS/MS.

In this work, a fast mycotoxin extraction (FaMEx) technique was developed for the rapid identification and quantification of carcinogenic ochratoxin-A (OTA) in food (coffee and tea) and agricultural soil samples using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) detection. The FaMEx technique advancement is based on two plastic syringes integrated setup for rapid extraction and its subsequent controlled clean-up process. In the extraction process, a 0.25-g sample and extraction solvent were added to the first syringe barrel for the vortex-based extraction. Then, the extraction syringe was connected to a clean-up syringe (pre-packed with C18, activated carbon, and MgSO4) with a syringe filter. Afterward, the whole set-up was placed in an automated programmable mechanical set-up for controlled elution. To enhance FaMEx technology performance, the various influencing sample pretreatment parameters were optimized. Furthermore, the developed FaMEx method indicated excellent linearity (0.9998 and 0.9996 for coffee/tea and soil) with highly sensitive detection (0.30 and 0.29 ng/mL for coffee/tea and soil) and quantification limits (1.0 and 0.96 for coffee/tea and soil), which is lower than the toxicity limit compliant with the European Union regulation for OTA (5 ng/g). The method showed acceptable relative recovery (84.48 to 100.59%) with <7.34% of relative standard deviation for evaluated real samples, and the matrix effects were calculated as <-13.77% for coffee/tea and -9.7 for soil samples. The obtained results revealed that the developed semi-automated FaMEx/UHPLC-MS/MS technique is easy, fast, low-cost, sensitive, and precise for mycotoxin detection in food and environmental samples.

Novel fast pesticides extraction (FaPEx) strategy coupled with UHPLC-MS/MS for rapid monitoring of emerging pollutant fipronil and its metabolite in food and environmental samples.

In this work, we demonstrated a new, environmental-friendly and effective sample preparation strategy named 'in-syringe-assisted fast pesticides extraction (FaPEx)' technique coupled with LC-MS/MS for the rapid identification and monitoring of emerging pollutant fipronil and its metabolite fipronil sulfone in chicken egg and environmental soil samples. FaPEx strategy comprising of two simple steps. Firstly, the sample was placed in the syringe and extracted using low-volume acetonitrile with NaCl and anhydrous MgSO4 salts. Secondly, the extractant was passed through in-syringe-based solid-phase extraction (SPE) kit containing cleanup sorbents and salt combinations (C18, primary secondary amine, and anhydrous MgSO4) for the cleanup process. Then, the obtained clean extractant was injected into LC-MS/MS for the quantification of target analytes. Various important parameters influencing the FaPEx performances, such as solvent type, salt type, salt amount, sorbent type, and amount, were examined and optimized. The method validation results showed excellent linearity with high correlation coefficients were ≥ 0.99. The estimated LODs were between 0.05-0.07 µg/kg, and LOQs ranged between 0.1-0.25 µg/kg for target analytes in both egg and soil sample matrices, and precision values were ≤7.90%. The developed method was applied to commercial chicken egg samples and environmental soil samples analysis. Spiked recoveries ranged between 88.75-110.91% for egg samples with RSDs ≤7.42% and 82.47-107.46% for soil samples with RSDs <7.37%. These results proved that the developed sample preparation method is a simple, fast, green, low-cost, and efficient method for the analysis of fipronil and its metabolites in food and environmental samples. Thus, this method can be applied as an alternative analytical methodology in routine and standard food and environmental testing laboratories.

Evaluation of optimal QuEChERS conditions of various food matrices for rapid determination of EU priority polycyclic aromatic hydrocarbons in various foods.

Optimal QuEChERS (quick, easy, cheap, effective, rugged and safe) conditions with good accuracy, repeatability and precision were established to rapidly extract the European Union (EU) priority polycyclic aromatic hydrocarbons (PAHs) from various food matrices (Category: Poultry and Meat, Fish and seafood, Grains, Soy beans and products, Root vegetables and Coffee). The QuEChERS conditions combined with the established high performance liquid chromatography-fluorescence detection conditions were used to rapidly determine the PAHs in 19 popular cooked foods in Taiwan and their corresponding original materials. These conditions also meet the EU and Taiwan Food and Drug Administration specifications. Charcoal grilled, gas stove grilled and smoked foods had higher PAHs contents, while fried and electric oven-baked/baked foods had lower PAHs contents. In addition to the effects of cooking methods, the contamination of original materials by PAHs in the environment should also have an important impact on the contents of PAHs in these cooked foods.

Comparison and establishment of appropriate methods to determine EU priority PAHs in charcoal-grilled chicken drumsticks with different treatments and their dietary risk assessments.

Three major polycyclic aromatic hydrocarbons (PAHs) analytical methods, gas chromatography-mass spectrometer (GC-MS), high performance liquid chromatography-photodiode-array detector (HPLC-PDA) and HPLC-fluorescence detector (FLD) were compared to analyze the European Union (EU) priority PAHs. In addition to the highest sensitivity, HPLC-FLD could be developed to rapidly determine the PAHs. A QuEChERS method was also established to rapidly extract the PAHs from chicken drumsticks. The method detection limits (0.004-0.25 ng/g), method quantification limits (0.01-0.75 ng/g), recoveries (67-114%), and coefficients of variations for intra- (1-15%) and inter- (1-21%) assays for the determination of the PAHs were in compliance with the EU and Taiwan Food and Drug Administration (TFDA) requirements. The PAHs contents in charcoal-grilled chicken drumsticks (without skin, with skin, and skin removal after processing) at different processing times were also determined by the developed methods. Although the risk assessment results for all samples showed a low level of concern, the presence of skin and long-term grilling had a greater impact on PAHs levels and dietary risks. Therefore, excessive processing should be avoided, and the skin of chicken drumsticks should be removed before and after charcoal-grilling to reduce the risk of PAHs intake.