Smartphone as a fluorescence detector for high-performance liquid chromatography.

BACKGROUND: Fluorescence detection is employed in high-performance liquid chromatography (HPLC) due to its high specificity and sensitivity. However, it is often limited by expensive components and bulkiness. Recently, advances in technology and electronics have led to the development of smartphones that can serve as portable recording, analysis, and monitoring tools. Smartphone-based detection provides advantages of cost effectiveness, rapid signal/data processing, and the display of results on a handhold monitor. The combination of smartphone-based detection with HPLC can offer unique features that are beneficial in overcoming limitations of commercial fluorescence detectors. (90) RESULTS: A miniaturized and low-cost HPLC fluorescence detector based on a smartphone is introduced for the detection of six fluorescent molecules. The smartphone is able to capture emitted fluorescence in video format while MATLAB code is used for data processing to provide chromatograms based on different detection channels. A custom designed double-channel flow cell was utilized to enable simultaneous detection of fluorescent compounds with different excitation wavelengths. The detector consists of a lab-made flow cell, monochromatic LEDs as the light source, 3D printed housing and connector box, fiber optic cables, and a smartphone. The effects of flow cell geometry, channel width and light slit diameter, as well as a comparison of different flow cell manufacturing techniques, are studied and discussed. The validated system was successfully applied to samples from diverse water sources, yielding spiking recoveries within the range of 91.7% and 109.7%. (141) SIGNIFICANCE: This study introduces the first smartphone-based fluorescence detector for HPLC with cost-effective and customizable flow cells, allowing for the simultaneous detection of fluorescent compounds with different excitation wavelengths and offering a potential solution for the analysis of co-eluting compounds. Beyond its user-friendly interface and low-cost, smartphone detection in HPLC provides tremendous opportunities in further miniaturizing chromatographic instrumentation while offering high sensitivity and can be expanded to other mechanisms of detection. (70).

Batch Scale Production of 3D Printed Extraction Sorbents Using a Low-Cost Modification to a Desktop Printer.

This study reports a simple modification to a commercial resin 3D printer that significantly reduces the amount of prepolymer material needed for the production of extraction sorbents. The modified printing platform is demonstrated in the printing of two imidazolium-based ionic liquid (IL) monomers. Two geometries resembling a blade-type polymeric ionic liquid (PIL) sorbent used in thin-film microextraction and a fiber-type sorbent used in solid-phase microextraction (SPME) were printed. The SPME PIL sorbents were used to extract 10 organic contaminants, including plasticizers, antimicrobial agents, UV filters, and pesticides, from water followed by high-performance liquid chromatographic (HPLC) analysis. To compare the extraction performance of the SPME sorbents, seven fibers printed with the same prepolymer composition from the same printing batch as well as different batches were evaluated. The results revealed highly reproducible extraction efficiencies for all tested sorbents with no statistical difference in their extraction performance. Method validation showed acceptable linearity (R2 > 0.92) for all analytes with limits of detection and limits of quantification ranging from 0.13 to 45 µg L-1 and 0.43 to 150 µg L-1, respectively.

Monitoring the liberation of volatile organic compounds during fused deposition modeling three dimensional printing using solid-phase microextraction coupled to gas chromatography/mass spectrometry.

Three-dimensional (3D) printers have gained tremendous popularity and are being widely used in offices, laboratories, and private homes. Fused deposition modeling (FDM) is among the most commonly used mechanisms by desktop 3D printers in indoor settings and relies on the extrusion and deposition of heated thermoplastic filaments, resulting in the liberation of volatile organic compounds (VOCs). With the growing use of 3D printers, concerns regarding human health have risen as the exposure to VOCs may cause adverse health effects. Therefore, it is important to monitor VOC liberation during printing and to correlate it to filament composition. In this study, VOCs liberated with a desktop printer were measured by solid-phase microextraction (SPME) combined with gas chromatography/mass spectrometry (GC/MS). SPME fibers featuring sorbent coatings of varied polarity were chosen for the extraction of VOCs liberated from acrylonitrile butadiene styrene (ABS), tough polylactic acid, and copolyester+ (CPE+) filaments. It was found that for all three filaments tested, longer print times resulted in a greater number of extracted VOCs. The ABS filament liberated the most VOCs while the CPE+ filaments liberated the fewest VOCs. Through the use of hierarchical cluster analysis and principal component analysis, filaments as well as fibers could be differentiated based on the liberated VOCs. This study demonstrates that SPME is a promising tool to sample and extract VOCs liberated during 3D printing under non-equilibrium conditions and can be used to aid in tentative identification of the VOCs when coupled to gas chromatography-mass spectrometry.

Digital Droplet Loop-Mediated Isothermal Amplification Featuring a Molecular Beacon Assay, 3D Printed Droplet Generation, and Smartphone Imaging for Sequence-Specific DNA Detection.

Nucleic acid detection is widely used in the amplification and quantitation of nucleic acids from biological samples. While polymerase chain reaction (PCR) enjoys great popularity, expensive thermal cyclers are required for precise temperature control. Loop-mediated isothermal amplification (LAMP) enables highly sensitive, rapid, and low-cost amplification of nucleic acids at constant temperatures. LAMP detection often relies on double-stranded DNA-binding dyes or metal indicators that lack sequence selectivity. Molecular beacons (MBs) are hairpin-shaped oligonucleotide probes whose sequence specificity in LAMP provides the capability of differentiating between single-nucleotide polymorphisms (SNPs). Digital droplet LAMP (ddLAMP) enables a large number of independent LAMP reactions to be performed and provides quantification of target DNA sequences. However, a major challenge with ddLAMP is the requirement of expensive droplet generators to form homogeneous microdroplets. In this study, we demonstrate for the first time that a three-dimensional (3D) printed droplet generation platform can be coupled to a LAMP assay featuring MBs as sequence-specific probes. The low-cost 3D printed droplet generator system was designed, and its customizability was demonstrated in the formation of monodisperse ddLAMP assay-in-oil microdroplets. Additionally, a smartphone-based imaging system is demonstrated to increase accessibility for point-of-care applications. The MB-ddLAMP assay is shown to discriminate between two SNPs at various amplification temperatures to afford a useful platform for sequence-specific, sensitive, and accurate DNA quantification. This work expands the utility of MBs to ddLAMP for quantitative studies in the detection of SNPs and exploits the customizability of 3D printing technologies to optimize the homogeneity, size, and volume of oil-in-water microdroplets.

Gold-Catalyzed Annulations of N-Propargyl Ynamides with Anthranils with Two Distinct Chemoselectivities.

Gold-catalyzed annulation of N-propargyl ynamides with anthranils can proceed by two distinct mechanisms. In the case of a terminal N-propargyl ynamide, its resulting α-imino gold carbene reacts with a tethered alkyne to generate a vinyl cation to enable hydrolysis, which ultimately yields a pyrrolo[2,3-b]quinoline derivative after treatment with p-toluenesulfonic acid. For an internal alkyne, its α-imino gold carbene reacts with a tethered alkyne via either a vinyl cation or an alkenylgold carbene; both paths ultimately lead to a 4-ketone-2-aminopyrrole derivative. Our mechanistic analysis indicates that water is a better nucleophile than anthranil for terminal ynamides, whereas water and anthranils are equally reactive for internal ynamides.

Gold-catalyzed N,O-functionalization of 1,4-diyn-3-ols with N-hydroxyanilines to form highly functionalized pyrrole derivatives.

This work describes new N,O-functionalization of 1,4-diyn-3-ols with N-hydroxyanilines to yield highly functionalized pyrrole derivatives. In a postulated mechanism, N-hydroxyaniline attacks the more electron-rich alkynes via regioselective N-attack to form unstable ketone-derived nitrones that react with their tethered alkynes via an intramolecular oxygen-transfer to form α-oxo gold carbenes. This new method is applicable to a short synthesis of a bioactive molecule, a PDE4 inhibitor.