Development of a Cation Exchange SPE-HILIC-MS/MS Method for the Determination of Ningnanmycin Residues in Tea and Chrysanthemum.

Ningnanmycin is a widely used antibiotic in agricultural production that effectively controls fungal and viral diseases in tea trees and chrysanthemums. The polarity characteristic of ningnanmycin has posed limitations on the development of robust detection methods, thereby hindering effective monitoring and control measures. By combining cation exchange solid phase extraction (SPE) with hydrophilic interaction chromatography tandem mass spectrometry (HILIC-MS/MS), we have effectively tackled the issue pertaining to the separation and retention of ningnanmycin. The average recoveries of ningnanmycin in green tea, black tea, and chrysanthemum were 77.3-82.0%, 80.1-81.5%, and 74.0-80.0%, respectively. The intraday and interday relative standard deviations (RSDs) were below and equal to 7.7%. Good linearity was observed in the concentration range of 1-1000 µg/L (R2 > 0.998). The limits of detection (LODs) ranged from 1.1 µg/kg to 7.1 µg/kg, and the limits of quantification (LOQs) ranged from 3.6 µg/kg to 23.7 µg/kg for ningnanmycin. These results indicate the good accuracy, repeatability, reproducibility, and sensitivity of the method. It is suitable for detecting ningnanmycin in tea and chrysanthemum.

Near-infrared-excitable acetylcholinesterase-activated fluorescent probe for sensitive and anti-interference detection of pesticides in colored food.

The development of a common and anti-interference acetylcholinesterase (AChE) inhibition assay for plant-originated food samples has been of great challenge because of the prevalent and strong signal interferences from natural pigments. Plant pigments normally exhibit non-negligible absorbance in the UV-visible region. As a result, the signals of a typical near-infrared (NIR) fluorescent probe could be disturbed through primary inner filter effect if it is excited by UV-visible light during plant sample analysis. In this work, an NIR-excitable AChE-activated fluorescent probe was biomimetically designed and synthesized. And the NIR-excitation strategy was utilized for the anti-interference detection of organophosphate and carbamate pesticides in colored samples with this probe. Sensitive and rapid response to AChE and pesticides was achieved due to the high affinity of the biomimetic recognition unit in the probe. The limits of detection for four representative pesticides including dichlorvos, carbofuran, chlorpyrifos and methamidophos reached 0.0186 µg/L, 2.20 µg/L, 12.3 µg/L and 13.6 µg/L, respectively. Most importantly, fluorescent response to pesticide contents could be accurately measured in the coexistence of different plant pigments by this probe, and the measured results showed completely irrelevance to the plant pigments and their colors. Taking advantage of such probe, the new developed AChE inhibition assay showed good sensitivity and anti-interference ability in the detection of organophosphate and carbamate pesticides in real samples.

A method for the rapid determination of pesticides coupling thin-layer chromatography and enzyme inhibition principles.

Herein, we developed a method coupling TLC and enzyme inhibition principles to rapidly detect OPs (dichlorvos, paraoxon and parathion). After the removal of the organic solvent from the samples using TLC and paper-based chips, the enzyme was added to the detection system. The results showed that the current method effectively reduced the effects of solvents on enzyme behavior. Moreover, the pigments could be successfully retained on TLC with 40% ddH2O/ACN solution (v/v) as a developing solvent. Additionally, the detection limits (LODs) were 0.002 µg/mL for dichlorvos, 0.006 µg/mL for paraoxon, and 0.003 µg/mL for parathion. Finally, the method was applied to spiked cabbage, cucumber, and spinach and showed good average recoveries ranging between 70.22% and 119.79%. These results showed that this paper-based chip had high sensitivity, precleaning, and elimination of organic solvent properties. Furthermore, it provides a valuable idea for sample pretreatment and rapid determination of pesticide residues in food.

Rapid measurement of total polyphenol content in tea by kinetic matching approach on microfluidic paper-based analytical devices.

In this work, a facile kinetic matching approach for total polyphenol content (TPC) measurement was developed based on the adoption of microfluidic paper-based analytical devices with symmetric channel distribution. A set of Folin-Ciocalteu reactions performed on the same paper chip were activated all at the same time through synchronized filling of sodium carbonate solution among individual channels. Gallic acid was found valid as a standard compound for kinetic matching measurement of tea samples. TPC of tea infusions was successfully measured within ten minutes without any complexed time control procedure needed. Under the optimized conditions, the new developed method showed good linearity in the TPC range of 10-100 mg/L (r > 0.9955) and the inter-chip precision was 5.6% (n = 11). The results measured with the new developed approach were in good agreement with those with the conventional FC assay.

A facile microfluidic paper-based analytical device for acetylcholinesterase inhibition assay utilizing organic solvent extraction in rapid detection of pesticide residues in food.

The incompatibility of most organic solvents with acetylcholinesterase (AChE) inhibition assay normally limits pesticide extraction efficiency in sample pretreatment, which might cause false negatives in real world sample assessment. Herein, a novel method has been developed for an improved AChE inhibition assay via organic solvent extraction combined spontaneous in situ solvent evaporation on microfluidic paper-based analytical devices. Enzyme pre-immobilization procedure was spared and AChE was added to the system after sampling step until a complete in-situ solvent evaporation process was performed on chip. IC50 levels of the six investigated organophosphate and carbamate pesticides indicated a completely eliminated influence of solvents on AChE behavior with the new method. Most importantly, analytical performances were significantly improved in food sample measurements. Reduction in matrix effect was observed when acetonitrile was adopted for lettuce sample pretreatment instead of water. Studies on different pesticides suggested a remarkably decreased discrimination effect on recoveries from sample pretreatment with the new developed method. The recovery level for phoxim spiked head lettuce samples reached (107.5 ± 14.2) %, in comparison with that of (18.6 ± 1.4) % from water-based extraction. Spiked water and apple juice samples with carbaryl concentration of as low as 0.02 mg L-1 were also successfully recognized with the present method by visual detection. This is the first report on direct sampling of organic extracts for AChE inhibition assay on-chip and it might provide a new perspective for real world sample assessments involving bio-reagents.

Residue pattern of polycyclic aromatic hydrocarbons during green tea manufacturing and their transfer rates during tea brewing.

Residues of polycyclic aromatic hydrocarbons (PAHs) in green tea and tea infusion were determined using gas chromatography-tandem mass spectrometry to study their dissipation pattern during green tea processing and infusion. Concentration and evaporation of PAHs during tea processing were the key factors affecting PAH residue content in product intermediates and in green tea. PAH residues in tea leaves increased by 2.4-3.1 times during the manufacture of green tea using the electric heating model. After correction to dry weight, PAH residue concentrations decreased by 33.5-48.4% during green tea processing because of PAH evaporation. Moreover, spreading and drying reduced PAH concentrations. The transfer rates of PAH residues from green tea to infusion varied from 4.6% to 7.2%, and PAH leaching was higher in the first infusion than in the second infusion. These results are useful for assessing exposure to PAHs from green tea and in formulating controls for the maximum residue level of PAHs in green tea.

Multiresidue Method for the Rapid Determination of Pesticide Residues in Tea Using Ultra Performance Liquid Chromatography Orbitrap High Resolution Mass Spectrometry and In-Syringe Dispersive Solid Phase Extraction.

A method based on in-syringe dispersive solid phase extraction (IS-D-SPE) and ultra performance liquid chromatography Orbitrap high resolution mass spectrometry for the multiresidue analysis of 117 pesticides in tea was developed. Full scan mode was acquired over an m/z range of 100-800 with Orbitrap resolution at 70000, followed by full scan/dd-MS2 mode for confirmation. The identification criteria of retention time and mass accuracy tolerance was ±0.20 min and ±5.0 ppm, respectively. MS/MS fragment ions obtained dd-MS2 were necessary to identify the pesticides with the same molecular mass weight. The IS-D-SPE technique involved a mixture of 200 mg PSA, 100 mg C18, and 15 mg multiwalled carbon nanotubes for the cleanup of tea matrix. Good linearity (R2 > 0.99) for 117 pesticides was obtained. Satisfactory recoveries in the range of 70-120% were obtained for 105 pesticides, while intraday and interday precisions were below 20%. Limits of quantification were generally 10 µg kg-1. Finally, this method was employed to analyze 117 pesticides in 70 tea samples.

Occurrence and Residue Pattern of Phthalate Esters in Fresh Tea Leaves and during Tea Manufacturing and Brewing.

The residues of 16 phthalate esters (PAEs) in fresh tea leaves and made tea were determined via gas chromatography-tandem mass spectrometry to study their distribution and degradation characteristics during tea planting and processing. Five PAEs were detected in all fresh tea leaves, and higher concentrations were detected in mature leaves. The distribution of PAEs in fresh tea leaves ranged from 69.7 to 2244.0 µg/kg. The degradative percentages of ∑5PAEs during green tea manufacturing ranged from 61 to 63% and were significantly influenced by the drying process. The transfer rates of PAEs-D4 ranged from 5.2 to 100.6%. PAEs with a high water solubility showed the highest transfer coefficient in the range of 91.8-100.6%, whereas PAEs with a high log Kow showed a low leaching efficiency below 11.9%. These results benefit the risk evaluation and establishment of a maximum residue limit for PAEs in tea.

Quantifying cell adhesion through impingement of a controlled microjet.

The impingement of a submerged, liquid jet onto a cell-covered surface allows assessing cell attachment on surfaces in a straightforward and quantitative manner and in real time, yielding valuable information on cell adhesion. However, this approach is insufficiently characterized for reliable and routine use. In this work, we both model and measure the shear stress exerted by the jet on the impingement surface in the micrometer-domain, and subsequently correlate this to jet-induced cell detachment. The measured and numerically calculated shear stress data are in good agreement with each other, and with previously published values. Real-time monitoring of the cell detachment reveals the creation of a circular cell-free area upon jet impingement, with two successive detachment regimes: 1), a dynamic regime, during which the cell-free area grows as a function of both the maximum shear stress exerted by the jet and the jet diameter; followed by 2), a stationary regime, with no further evolution of the cell-free area. For the latter regime, which is relevant for cell adhesion strength assessment, a relationship between the jet Reynolds number, the cell-free area, and the cell adhesion strength is proposed. To illustrate the capability of the technique, the adhesion strength of HeLa cervical cancer cells is determined ((34 ± 14) N/m(2)). Real-time visualization of cell detachment in the dynamic regime shows that cells detach either cell-by-cell or by collectively (for which intact parts of the monolayer detach as cell sheets). This process is dictated by the cell monolayer density, with a typical threshold of (1.8 ± 0.2) × 10(9) cells/m(2), above which the collective behavior is mostly observed. The jet impingement method presents great promises for the field of tissue engineering, as the influence of both the shear stress and the surface characteristics on cell adhesion can be systematically studied.

In vitro development of donated frozen-thawed human embryos in a prototype static microfluidic device: a randomized controlled trial.

OBJECTIVE: To compare the development of human embryos in microfluidic devices with culture in standard microdrop dishes, both under static conditions. DESIGN: Prospective randomized controlled trial. SETTING: In vitro fertilization laboratory. PATIENT(S): One hundred eighteen donated frozen-thawed human day-4 embryos. INTERVENTION(S): Random allocation of embryos that fulfilled the inclusion criteria to single-embryo culture in a microfluidics device (n = 58) or standard microdrop dish (n = 60). MAIN OUTCOME MEASURE(S): Blastocyst formation rate and quality after 24, 28, 48, and 72 hours of culture. RESULT(S): The percentage of frozen-thawed day-4 embryos that developed to the blastocyst stage did not differ significantly in the standard microdrop dishes and microfluidic devices after 28 hours of culture (53.3% vs. 58.6%) or at any of the other time points. The proportion of embryos that would have been suitable for embryo transfer was comparable after 28 hours of culture in the control dishes and microfluidic devices (90.0% vs. 93.1%). Furthermore, blastocyst quality was similar in the two study groups. CONCLUSION(S): This study shows that a microfluidic device can successfully support human blastocyst development in vitro under static culture conditions. Future studies need to clarify whether earlier stage embryos will benefit from the culture in microfluidic devices more than the tested day-4 embryos because many important steps in the development of human embryos already take place before day 4. Further improvements of the microfluidic device will include parallel culture of single embryos, application of medium refreshment, and built-in sensors. DUTCH TRIAL REGISTRATION NUMBER: NTR3867.

Preparation of micro gold devices on poly(dimethylsiloxane) chips with region-selective electroless plating.

A novel protocol for fabrication of micro gold devices on poly(dimethylsiloxane) (PDMS) substrates was developed on the basis of region-selective electroless plating. The layout of a micro gold device was first photochemically patterned onto the PDMS surface through a UV induced poly(acrylic acid) (PAA) grafting process. The carboxylic moieties on the grafted PAA served as the scaffold for a series of wet chemical reactions that led to the immobilization of gold nanoparticles in the UV-exposed region, where electroless plating then occurred under the catalysis of the nanoparticles. Gold devices fabricated with such a protocol could tolerate the Scotch tape test and survive in a repeated bending-straightening test. They also showed good stability in acidic and alkaline solutions, possessed almost the same electrochemical properties as a standard gold disk electrode, and allowed thiol-compounds to form a perfect self-assembled monolayer on their surfaces. The fabricated micro gold electrode was demonstrated to be suitable as the integrated amperometric detection element in a full PDMS micro electrophoresis chip.

Fracture mechanism of metal electrode integrated on a chip and fabrication of a poly(ethylene terephthalate) electrophoresis microchip.

Thermal bonding is an important technique to fabricate polymer electrophoresis microchip. However, the metal electrodes deposited on polymer substrate can readily fracture during the thermal bonding. In this paper, poly(ethylene terephthalate) (PET) was exploited to fabricate the electrophoresis microchip with an integrated gold electrode for amperometric detection. The fracture of the gold electrode was studied through FEA (finite element analysis) simulations, the potentially risk positions on the electrode were shown. The calculation results were tested by bonding experiments and were proven to be consistent with the experiments. Besides, an optimal bonding temperature for PET chip was also presented based on FEA simulations and bonding experiments. Considering the low surface properties of PET, oxygen plasma-assisted thermal bonding technique was used to enhance bonding. Upon treated for 150 s, the PET substrates could be thermally bonded at 62 degrees C without electrode fracture. The fabricated PET chips were demonstrated for detection of standard glucose solution. Satisfactory reproducibility was achieved, and the RSD values of peak height and migration time of the PET CE chips were 0.51% and 2.17%, respectively.

Modification of amorphous poly(ethylene terephthalate) surface by UV light and plasma for fabrication of an electrophoresis chip with an integrated gold microelectrode.

Amorphous poly(ethylene terephthalate) (PET), which possess a low softening temperature (T(s)=75 degrees C), was exploited to fabricate the electrophoresis chip with an integrated gold electrode for amperometric detection, with emphases being focused on the PET surface modification via UV light and air plasma. Both UV irradiation and plasma treatment were found to be able to improve the surface wettability, enhance the supported electroosmotic flow (EOF), and increase thermal bonding strength of PET sheets, with the latter being more efficient and less time-consuming than the former in the surface modification. Upon treated with plasma for 2 min, the PET sheets could be thermally bonded at 65 degrees C. T-peer test showed that the bonding strength increased from 10 g/cm for native PET sheets to 1250 g/cm for the plasma treated sheets when chips were bonded at the softening point, Attenuated-total-internal-reflection spectrum showed that, after being exposed to the UV light, carboxylic groups site-selectively formed in the UV-exposed region on PET surface. These UV-induced carboxylic groups were further utilized as the scaffold for preparation of micro-gold electrode via electroless gold plating. By using this established UV-directed electroless plating and the plasma-assisted thermal bonding techniques, the full PET electrophoresis chip with an integrated micro-gold electrode could be fabricated in common chemistry laboratory without the need of clean rooms. The fabricated PET chips were demonstrated for separation and detection of model analytes of dopamine (DA) and catechol (CA). Satisfactory resolution of the two analytes was achieved within 40s, and detection limits of 0.87 microM and 1.28 microM for DA and CA were obtained, respectively.