Chromium-based metal organic framework for pipette tip micro-solid phase extraction: an effective approach for determination of methyl and propyl parabens in wastewater and shampoo samples

Background A chromium-based metal organic framework was synthesized and employed as an efficient sorbent for pipette tip micro-solid phase extraction and preconcentration of parabens from wastewater and shampoo samples up to sub-ppb level before their spectrophotometric analysis. Results Factors affecting preconcentration including volume and type of solvent, amount of sorbent, number of extraction, and volume and pH of samples were optimized employing one-variable-at-a-time and response surface methodology. Obtained analytical characteristics of the method proves its usefulness for analysis of real samples. Linear range of the method for parabens was 1.0–200.0 μg/L. Detection limit of the protocol was 0.24 µg/L for propyl paraben and 0.25 µg/L for methyl paraben. Reproducibility of the protocol defined as % RSD was better than 5.78%. Synthesized adsorbent can be re-used for at least 20 extractions. Conclusion The method showed a good detection limit and precision for determination of methyl- and propyl-paraben in wastewater and shampoo samples.

Pipette Tip Solid-Phase Extraction Combined with Fluorescence Spectroscopy for Determination of Selenium in Green Tea Samples

In this study, the custom-built pipette-tip solid phase extraction (PT-SPE), was used for pre-concentrating the selenium (Se) in green tea samples. The PT-SPE was prepared using a pipette tip and C18 as the adsorbent that was placed between glass wool. This method afforded several advantages in terms of efficiency of extraction time (8 min), sample (2 mL), and solvent (1 mL) volumes with less wastage of solvent. Under optimal conditions of extraction, this method yielded good linearity (r2=0.998) for the Se calibration curve and good limit of detection (0.2 μg/L) and quantification (0.6 μg/L). The standard addition method was used to remove the matrix effect. The recovery percentages of tea samples spiked with different Se concentrations ranged from 97.7 to 106.0%, indicating that the proposed method would be suitable for the determination of Se in green tea samples.

SDS-PAGE gel electrophoresis v2

SDS-PAGE gel electrophoresis protocol for analyzing samples from plant leaf tissue via immunofluorescence. In this protocol no Coomassie blue is added to samples, the reason is that this interferes with the fluorescent signal during immunoblot. Instead, samples have already been prepared in Laemmli buffer (minus coomassie, protein extraction procedure), the leading edge of samples can be visualized due to the presence of chlorophyll. Note - When using 15 well, 0.75 mm comb, try to limit the volume loaded to 10 μL to minimize the risk of spillover of protein between wells. - Ensure that accurate volume is pipetted by removing sample stuck to the outside of the pipette tip by wiping the tip on the rim of the sample tube to remove any residual liquid. Literature: http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_6040.pdfhttps://www.bio-rad.com/webroot/web/pdf/lsr/literature/10026447.pdf

A linear programming-based strategy to save pipette tips in automated DNA assembly

Laboratory automation and mathematical optimisation are key to improving the efficiency of synthetic biology research. While there are algorithms optimising the construct designs and synthesis strategies for DNA assembly, the optimisation of how DNA assembly reaction mixes are prepared remains largely unexplored. Here, we focus on reducing the pipette tip consumption of a liquid-handling robot as it delivers DNA parts across a multi-well plate where several constructs are being assembled in parallel. We propose a linear programming formulation of this problem based on the capacitated vehicle routing problem, along with an algorithm which applies a linear programming solver to our formulation, hence providing a strategy to prepare a given set of DNA assembly mixes using fewer pipette tips. The algorithm performed well in randomly generated and real-life scenarios concerning several modular DNA assembly standards, proving capable of reducing the pipette tip consumption by up to 61% in large-scale cases. Combining automatic process optimisation and robotic liquid-handling, our strategy promises to greatly improve the efficiency of DNA assembly, either used alone or in combination with other algorithmic methods.

Recent Advances in Molecular-Imprinting-Based Solid-Phase Extraction of Antibiotics Residues Coupled With Chromatographic Analysis

Abuse and residues of antibiotics cause great harm to organisms and the environment. Appropriate sample pretreatment is usually required for sensitive determination, because of the low content presence of a variety of antibiotics in complicated matrices. Molecular-imprinting-based solid-phase extraction (MISPE) has been widely used for sample pretreatment of antibiotics, using molecularly imprinted polymers (MIPs) as adsorbents. Herein, we comprehensively review the recent advances of MISPE of antibiotics, followed by chromatographic analysis. Various solid-phase extraction (SPE) modes based on MIPs are briefly introduced, such as conventional SPE, dispersive SPE, magnetic SPE, matrix solid-phase dispersion, and pipette-tip SPE. Then, several emerging preparation techniques for antibiotics MIPs are summarized including surface imprinting, nanoimprinting, living/controlled radical polymerization, multitemplate imprinting, multifunctional monomer imprinting, and dummy template imprinting. Subsequently, applications of MISPE to analysis of a variety of antibiotics residues since 2018 are overviewed, including sulfonamides, quinolones, tetracycline, and others. Finally, the preparation and application of antibiotics MIPs are prospected.