Field-deployable Devices Research Articles

Turnkey photonic flywheel in a microresonator-filtered laser

Dissipative Kerr soliton (DKS) microcomb has emerged as an enabling technology that revolutionizes a wide range of applications in both basic science and technological innovation. Reliable turnkey operation with sub-optical-cycle and sub-femtosecond timing jitter is key to the success of many intriguing microcomb applications at the intersection of ultrafast optics and microwave electronics. Here we propose an approach and demonstrate the first turnkey Brillouin-DKS frequency comb to the best of our knowledge. Our microresonator-filtered laser design offers essential benefits, including phase insensitivity, self-healing capability, deterministic selection of the DKS state, and access to the ultralow noise comb state. The demonstrated turnkey Brillouin-DKS frequency comb achieves a fundamental comb linewidth of 100 mHz and DKS timing jitter of 1 femtosecond for averaging times up to 56 μs. The approach is universal and generalizable to various device platforms for user-friendly and field-deployable comb devices.

Nondestructive and Rapid Screening of Aflatoxin-Contaminated Single Peanut Kernels Using Field-Portable Spectroscopy Instruments (FT-IR and Raman).

A nondestructive and rapid classification approach was developed for identifying aflatoxin-contaminated single peanut kernels using field-portable vibrational spectroscopy instruments (FT-IR and Raman). Single peanut kernels were either spiked with an aflatoxin solution (30 ppb-400 ppb) or hexane (control), and their spectra were collected via Raman and FT-IR. An uHPLC-MS/MS approach was used to verify the spiking accuracy via determining actual aflatoxin content on the surface of randomly selected peanut samples. Supervised classification using soft independent modeling of class analogies (SIMCA) showed better discrimination between aflatoxin-contaminated (30 ppb-400 ppb) and control peanuts with FT-IR compared with Raman, predicting the external validation samples with 100% accuracy. The accuracy, sensitivity, and specificity of SIMCA models generated with the portable FT-IR device outperformed the methods in other destructive studies reported in the literature, using a variety of vibrational spectroscopy benchtop systems. The discriminating power analysis showed that the bands corresponded to the C=C stretching vibrations of the ring structures of aflatoxins were most significant in explaining the variance in the model, which were also reported for Aspergillus-infected brown rice samples. Field-deployable vibrational spectroscopy devices can enable in situ identification of aflatoxin-contaminated peanuts to assure regulatory compliance as well as cost savings in the production of peanut products.

Electrocatalytic Palladium Nanoclusters as Versatile Indicators of Bioassays: Rapid Electroanalytical Detection of SARS-CoV-2 by Reverse Transcription Loop-Mediated Isothermal Amplification.

Quantitative polymerase chain reaction (qPCR) is considered the gold standard for pathogen detection. However, improvement is still required, especially regarding the possibilities of decentralization. Apart from other reasons, infectious diseases demand on-site analysis to avoid pathogen spreading and increase treatment efficacy. In this paper, the detection of SARS-CoV-2 is carried out by reverse transcription loop-mediated isothermal amplification, which has the advantage of requiring simple equipment, easily adaptable to decentralized analysis. It is proposed, for the first time, the use of palladium nanoclusters (PdNCs) as indicators of the amplification reaction at end point. The pH of the medium decreases during the reaction and, in turn, a variation in the catalytic activity of PdNCs on the oxygen reduction reaction (ORR) can be electrochemically observed. For the detection, flexible and small-size screen-printed electrodes can be premodified with PdNCs, which together with the use of a simple and small electrochemical equipment would greatly facilitates their integration in field-deployable devices. This would allow a faster detection of SARS-CoV-2 as well as of other future microbial threats after an easy adaptation.

Plasmonic/magnetic nanoarchitectures: From controllable design to biosensing and bioelectronic interfaces

Controllable design of the nanocrystal-assembled plasmonic/magnetic nanoarchitectures (P/MNAs) inspires abundant methodologies to enhance light-matter interactions and control magnetic-induced effects by means of fine-tuning the morphology and ordered packing of noble metallic or magnetic building blocks. The burgeoning development of multifunctional nanoarchitectures has opened up broad range of interdisciplinary applications including biosensing, in vitro diagnostic devices, point-of-care (POC) platforms, and soft bioelectronics. By taking advantage of their customizability and efficient conjugation with capping biomolecules, various nanoarchitectures have been integrated into high-performance biosensors with remarkable sensitivity and versatility, enabling key features that combined multiplexed detection, ease-of-use and miniaturization. In this review, we provide an overview of the representative developments of nanoarchitectures that being built by plasmonic and magnetic nanoparticles over recent decades. The design principles and key mechanisms for signal amplification and quantitative sensitivity have been explored. We highlight the structure-function programmability and prospects of addressing the main limitations for conventional biosensing strategies in terms of accurate selectivity, sensitivity, throughput, and optoelectronic integration. State-of-the-art strategies to achieve affordable and field-deployable POC devices for early multiplexed detection of infectious diseases such as COVID-19 has been covered in this review. Finally, we discuss the urgent yet challenging issues in nanoarchitectures design and related biosensing application, such as large-scale fabrication and integration with portable devices, and provide perspectives and suggestions on developing smart biosensors that connecting the materials science and biomedical engineering for personal health monitoring.

Noble Metal Nanoparticle Biosensors: From Fundamental Studies toward Point-of-Care Diagnostics.

Noble metal nanoparticles (NMNPs) have become firmly established as effective agents to detect various biomolecules with extremely high sensitivity. This ability stems from the collective oscillation of free electrons and extremely large electric field enhancement under exposure to light, leading to various light-matter interactions such as localized surface plasmon resonance (LSPR) and surface-enhanced Raman scattering. A remarkable feature of NMNPs is their customizability by mechanisms such as particle etching, growth, and aggregation/dispersion, yielding distinct color changes and excellent opportunities for colorimetric biosensing in user-friendly assays and devices. They are readily functionalized with a large variety of capping agents and biomolecules, with resultant bioconjugates often possessing excellent biocompatibility, which can be used to quantitatively detect analytes from physiological fluids. Furthermore, they can possess excellent catalytic properties that can achieve significant signal amplification through mechanisms such as the catalytic transformation of colorless substrates to colored reporters. The various excellent attributes of NMNP biosensors have put them in the spotlight for developing high-performance in vitro diagnostic (IVD) devices that are particularly well-suited to mitigate the societal threat that infectious diseases pose. This threat continues to dominate the global health care landscape, claiming millions of lives annually. NMNP IVDs possess the potential to sensitively detect infections even at very early stages with affordable and field-deployable devices, which will be key to strengthening infectious disease management. This has been the major focal point of current research, with a view to new avenues for early multiplexed detection of infectious diseases with portable devices such as smartphones, especially in resource-limited settings.In this Account, we provide an overview of our original inspiration and efforts in NMNP-based assay development, together with some more sophisticated IVD assays by ourselves and many others. Our work in the area has led to our recent efforts in developing IVDs for high-profile infectious diseases, including Ebola and HIV. We emphasize that integration with digital platforms represents an opportunity to establish and efficiently manage widespread testing, tracking, epidemiological intelligence, and data sharing backed by community participation. We highlight how digital technologies can address the limitations of conventional diagnostic technologies at the point of care (POC) and how they may be used to abate and contain the spread of infectious diseases. Finally, we focus on more recent integrations of noble metal nanoparticles with Raman spectroscopy for accurate, noninvasive POC diagnostics with improved sensitivity and specificity.

Detection of pks Island mRNAs Using Toehold Sensors in Escherichia coli.

Synthetic biologists have applied biomolecular engineering approaches toward the goal of novel biological devices and have shown progress in diverse areas of medicine and biotechnology. Especially promising is the application of synthetic biological devices towards a novel class of molecular diagnostics. As an example, a de-novo-designed riboregulator called toehold switch, with its programmability and compatibility with field-deployable devices showed promising in vitro applications for viral RNA detection such as Zika and Corona viruses. However, the in vivo application of high-performance RNA sensors remains challenging due to the secondary structure of long mRNA species. Here, we introduced ‘Helper RNAs’ that can enhance the functionality of toehold switch sensors by mitigating the effect of secondary structures around a target site. By employing the helper RNAs, previously reported mCherry mRNA sensor showed improved fold-changes in vivo. To further generalize the Helper RNA approaches, we employed automatic design pipeline for toehold sensors that target the essential genes within the pks island, an important target of biomedical research in connection with colorectal cancer. The toehold switch sensors showed fold-changes upon the expression of full-length mRNAs that apparently depended sensitively on the identity of the gene as well as the predicted local structure within the target region of the mRNA. Still, the helper RNAs could improve the performance of toehold switch sensors in many instances, with up to 10-fold improvement over no helper cases. These results suggest that the helper RNA approaches can further assist the design of functional RNA devices in vivo with the aid of the streamlined automatic design software developed here. Further, our solutions for screening and stabilizing single-stranded region of mRNA may find use in other in vivo mRNA-sensing applications such as cas13 crRNA design, transcriptome engineering, and trans-cleaving ribozymes.

Portable and Autonomous Device for Real-time Colorimetric Detection: Validation for Phosphorous and Nitrite Detection

Application of autonomous devices has disrupted routine digital monitoring in all the domains specifically in the fields of environmental monitoring, agriculture and allied sector, health care, and medical diagnostics. These modern autonomous field-deployable devices facilitate reliable testing, data acquisition, and efficient and immediate data transmission. Among various popular detection methods, colorimetric detection method is capable of detecting environmental samples with very low concentration with a high level of precision, are cost-effective, and provide ease-of-use. Hence, in this work, a portable, and autonomous device for real-time detection was designed using commercially available light-emitting diode (LED) and a photodiode, having peak response at a wavelength at which the chromophore exhibits absorption, were used to build the colorimetric detection device. The developed device is a portable, highly sensitive, low-cost, and low-power-consuming colorimetric detection platform integrated with Internet of Things (IoT) and Bluetooth modules for online and offline data transmission, respectively. The device functionality was further enhanced by adding other important attributes such as data storage, retrieval, and real-time monitoring capabilities. For the proof-of-concept, phosphate and nitrite samples were tested colorimetrically. Phosphorous was detected in the linear range of 10–100 μM with the limit of detection (LOD) being 0.074 ppm, while nitrite was detected in a linear range of 5–30 μM with LOD of 0.06 ppm. By suitably modifying the optoelectronic components, the developed colorimetric detection platform can be made potentially amenable to sense a variety of analytes.

High-energy single-longitudinal-mode mid-infrared optical parametric amplifier seeded with sheet optical parametric oscillator

Mid-infrared (MIR) radiation is essential for remote gas sensing, ranging, and lidar applications, where high pulse energy and narrow linewidth are the keys to the high sensitivity over long distance. However, complex optical and electronic locking schemes are normally required to achieve both features at the same time. Here, we demonstrate pulse-pumped single-longitudinal-mode (SLM) MIR generation using a microresonator seed in the form of a sheet optical parametric oscillator (SOPO). The SOPO features a sub-coherence-length thickness of 400 µm, which enables SLM and high-energy oscillation using cavity phase matching. Its output around 1.55 µm is seeded into an optical parametric amplifier and locks the MIR output in SLM at 3.38 µm. In a simple and compact setup, up to 21% conversion efficiency and 22% slope efficiency are measured with a MIR output energy of 54 µJ. This SLM MIR source with the SOPO seed greatly reduces the system size and is compatible for further integration for field-deployable devices and thus has broad applications in the field of remote gas sensing.

Quantification of Curcuminoids in Turmeric Using Visible Reflectance Spectra and a Decision-Tree Based Chemometric Approach

For quantification of curcumin content in turmeric, a low-cost multivariate-analysis-based sensing system is desired. It can be realized by exploiting the spectra in the visible region, which enables the use of off-the-shelf, relatively inexpensive light sources and detectors. To address this, we propose a novel decision-tree method for improved prediction accuracy. Two sets of models with PLSR algorithm are developed with the measured reflectance spectra from 66 turmeric samples in the range of 360–750 nm, and their respective curcuminoids content are quantified by HPLC. A suite of a coarse-model for initial prediction of turmeric samples in the broad range of 1%–4%, and five finer-models for subsequent prediction (in the ranges 1%–2%, 2%–3%, 3%–4%, 1.5%–2.5%, and 2.5%–3.5%) constitute the proposed decision-tree approach. The method’s efficacy is substantiated from an improved coefficient of determination (R 2) for the finer models (0.90–0.96) as compared to the coarse-model’s 0.92. This is further corroborated with lower RMSECV of 0.06–0.13 and an RMSEP of 0.15–0.25 for finer models, as compared to 0.219 and 0.45 for the coarse model, respectively. Testing reveals that the method results in 46% reduction in prediction error. Realization of a robust prediction approach in the visible range sets the stage for the development of cost-effective field-deployable devices for on-site measurement of curcumin.

Going beyond the Debye Length: Overcoming Charge Screening Limitations in Next-Generation Bioelectronic Sensors.

Electronic biosensors are a natural fit for field-deployable diagnostic devices because they can be miniaturized, mass produced, and integrated with circuitry. Unfortunately, progress in the development of such platforms has been hindered by the fact that mobile ions present in biological samples screen charges from the target molecule, greatly reducing sensor sensitivity. Under physiological conditions, the thickness of the resulting electric double layer is less than 1 nm, and it has generally been assumed that electronic detection beyond this distance is virtually impossible. However, a few recently described sensor design strategies seem to defy this conventional wisdom, exploiting the physics of electrical double layers in ways that traditional models do not capture. In the first strategy, charge screening is decreased by constraining the space in which double layers can form. The second strategy uses external stimuli to prevent double layers from reaching equilibrium, thereby effectively reducing charge screening. In this Perspective, we describe these relatively new concepts and offer theoretical insights into mechanisms that may enable electronic biosensing beyond the Debye length. If these concepts can be further developed and translated into practical electronic biosensors, we foresee exciting opportunities for the next generation of diagnostic technologies.

All-Fiber Hyperparametric Generation Based on a Monolithic Fiber Fabry–Pérot Microresonator

Hyperparametric oscillation is essential for coherent optical signal generation in a broad wavelength range. Integration in a compact system, such a broadband light source, is of special interest for practical applications requiring field-deployable spectroscopy devices. Here we demonstrate an all-fiber hyperparametric oscillation source based on four-wave mixing in a high-Q fiber Fabry–Pérot (FFP) microresonator. Assisted by the Raman effect, the generated optical signal spans over 400 nm with fine line-to-line spacing of 667 MHz. The compatibility of this FFP microresonator enables a robust and reliable all-fiber system through a splicing technique and fiber connectors. Such a plug-and-play platform is convenient and efficient for broad applications in optical communications and spectroscopy.

OPAA/fluoride biosensor chip towards field detection of G-type nerve agents

Sensor technology, particularly disposable, field-deployable devices offer considerable promise for addressing major security demands, in general, and in particular the urgent needs for selective and rapid field detection of highly toxic G-type organophosphate (OP) nerve agents. Herein, we report on the first example of a disposable electrochemical biosensor chip for decentralized field detection of diisopropyl fluorophosphate (DFP), as the simulant compound of G-type nerve agents (e.g., Sarin and Soman), based on the integration of an all-solid-state fluoride potentiometric transducer with the enzyme organophosphorus acid anhydrolase (OPAA) in connection to a hand-held potentiometric analyzer. Such solid-contact fluoride ion-selective electrodes were realized by comparing different fluoride ionophores to select the bis(fluorodioctylstannyl)methane molecule, along with systematic optimization of the experimental parameters. This first solid-contact fluoride detection chip displayed a favorable analytical performance, with near-Nernstian behavior at neutral pH in different buffers, high selectivity against various anions along with excellent reproducibility. Combining such desirable merits of the new fluoride sensor with the selective biocatalytic hydrolysis via an immobilized OPAA, to break the PF bond of G-type neurotoxins and release detectable fluoride ions, thereby enabled an attractive analytical performance. Both the fluoride and the nerve-agent recognition layers have thus been systematically optimized. The resulting biosensor chip thus displays a fast, sensitive and selective response towards DFP (including efficient discrimination against OP pesticides), along with a wide dynamic range of 250–3000 μM. Key challenges towards developing the nerve-agent detection chip were carefully addressed, including the pH incompatibility of the enzymatic and detection processes. By providing effective discrimination against common OP pesticides and minimizing related false alarms, the resultant disposable potentiometric biochip platform is expected to provide an instantaneous reliable warning of chemical attack and exposure towards a timely countermeasure action, and hence, would dramatically advance the protection of our military personnel.

Miniaturization of optical sensors and their potential for high-throughput screening of foods

Molecular fingerprinting technology has evolved from bulky laboratory benchtop instrumentation to field-deployable devices driven by advances in semiconductor and photonic technologies. The ongoing miniaturization of vibrational spectroscopy equipment has revolutionized the food industry by allowing on-site and real-time monitoring of food products and production processes to ensure quality and safety. The development of spectral libraries to fingerprint contaminants has provided unique detection capabilities to ensure consumer’s product safety. Commercialization of handheld and ruggedized instrumentation for field deployment is enabling little or no sample preparation requirement, non-contact and non-destructive capabilities. Testing done as close to the original source would permit detecting risks before an ingredient has been diluted or combined with other ingredients. By producing a characteristic chemical ‘fingerprint’ with unique signature profiles, miniaturized molecular spectroscopy techniques combined with chemometric analysis have positioned as viable ‘green’ alternatives for field applications allowing phenotyping, quality assurance, authentication, and detection of adulteration and contaminants in foods. Contrary to NMR and mass spectrometry (MS) techniques, vibrational spectroscopy provides selectivity and specificity for screening raw materials without requiring costly instrumentation, labor-intensive and complex sample pretreatment, well-trained technicians to operate the instrumentation and are particularly amenable to be implemented for quality control in the field, manufacturing facilities, and grocery stores. In this review, enabling technologies for further miniaturization of vibrational spectroscopy equipment and their applications to the food industry are presented.

Contributions and Challenges of High Throughput qPCR for Determining Antimicrobial Resistance in the Environment: A Critical Review.

Expansion in whole genome sequencing and subsequent increase in antibiotic resistance targets have paved the way of high throughput qPCR (HT-qPCR) for analyzing hundreds of antimicrobial resistance genes (ARGs) in a single run. A meta-analysis of 51 selected studies is performed to evaluate ARGs abundance trends over the last 7 years. WaferGenTM SmartChip is found to be the most widely used HT-qPCR platform among others for evaluating ARGs. Up till now around 1000 environmental samples (excluding biological replicates) from different parts of the world have been analyzed on HT-qPCR. Calculated detection frequency and normalized ARGs abundance (ARGs/16S rRNA gene) reported in gut microbiome studies have shown a trend of low ARGs as compared to other environmental matrices. Disparities in the HT-qPCR data analysis which are causing difficulties to researchers in precise interpretation of results have been highlighted and a possible way forward for resolving them is also suggested. The potential of other amplification technologies and point of care or field deployable devices for analyzing ARGs have also been discussed in the review. Our review has focused on updated information regarding the role, current status and future perspectives of HT-qPCR in the field of antimicrobial resistance.

Optical Flow Cell for Measuring Size, Velocity and Composition of Flowing Droplets

Here an optical flow cell with two light paths is reported that can accurately quantify the size and velocity of droplets flowing through a microchannel. The flow cell can measure the time taken for droplets to pass between and through two conjoined light paths, and thereby is capable of measuring the velocities (0.2–5.45 mm/s) and sizes of droplets (length > 0.8 mm). The composition of the droplet can also be accurately quantified via optical absorption measurements. The device has a small footprint and uses low-powered, low-cost components, which make it ideally suited for use in field-deployable and portable analytical devices.

Simple reagent storage in polyester-paper hybrid microdevices for colorimetric detection

A challenge in developing microfluidic devices for fully portable and automated field analyses is the ability to integrate and store reagents in a stable form while maintaining a low cost for the system. Paper-based field-deployable devices are currently the most extensively explored platforms for field use; however, centrifugal devices offer a means for heightened device automation and portability. The basis for many field devices is chemical sensing via colorimetric reactions. Here, we describe the development of polyester-paper hybrid microfluidic devices using a fabrication method, compounded from the print, cut and laminate (PCL) technique, that allows for simple yet effective dry reagent storage for enhanced use with centrifugal or polymeric-based devices. We utilized various proof-of-principle colorimetric assays for multiple applications, including the analysis of total protein, serum albumin, cocaine, 2,4,6-trinitrotoluene (TNT) and iron content. Localized reagent integration facilitated by paper punches allowed for both reagent stability prior to reaction for storage, color stability post-reaction for image analysis, and was compatible for multi-step analyses. Qualitative and quantitative results were achieved with the broad range of sample applications trialed here, while still maintaining the ability for downstream processing.

One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV.

Due to the limitation of rapid development of specific antiviral drug or vaccine for novel emerging viruses, an accurate and rapid diagnosis is a key to manage the virus spread. We developed an efficient and rapid method with high specificity for the Middle East Respiratory Syndrome coronavirus (MERS-CoV), based on one-pot reverse transcription loop-mediated isothermal amplification (one-pot RT-LAMP). A set of six LAMP primers [F3, B3, FIP, BIP, LF (Loop-F), and LB (Loop-B)] were designed using the sequence of nucleocapsid (N) gene with optimized RT-LAMP enzyme conditions: 100 U M-MLV RTase and 4 U Bst polymerase, implying that the reaction was able to detect four infectious viral genome copies of MERS-CoV within a 60 min reaction time period. Significantly, EvaGreen dye has better signal read-out properties in one-pot RT-LAMP reaction and is more compatible with DNA polymerase than SYBR green I. Isothermally amplified specific N genes were further evaluated using field-deployable microchamber devices, leading to the specific identification of as few as 0.4 infectious viral genome copies, with no cross-reaction to the other acute respiratory disease viruses, including influenza type A (H1N1 and H3N2), type B, human coronavirus 229E, and human metapneumovirus. This sensitive, specific and feasible method provides a large-scale technical support in emergencies, and is also applied as a sample-to-detection module in Point of Care Testing devices.

Low cost audiovisual playback and recording triggered by radio frequency identification using Raspberry Pi.

Playbacks of visual or audio stimuli to wild animals is a widely used experimental tool in behavioral ecology. In many cases, however, playback experiments are constrained by observer limitations such as the time observers can be present, or the accuracy of observation. These problems are particularly apparent when playbacks are triggered by specific events, such as performing a specific behavior, or are targeted to specific individuals. We developed a low-cost automated playback/recording system, using two field-deployable devices: radio-frequency identification (RFID) readers and Raspberry Pi micro-computers. This system detects a specific passive integrated transponder (PIT) tag attached to an individual, and subsequently plays back the stimuli, or records audio or visual information. To demonstrate the utility of this system and to test one of its possible applications, we tagged female and male tree swallows (Tachycineta bicolor) from two box-nesting populations with PIT tags and carried out playbacks of nestling begging calls every time focal females entered the nestbox over a six-hour period. We show that the RFID-Raspberry Pi system presents a versatile, low-cost, field-deployable system that can be adapted for many audio and visual playback purposes. In addition, the set-up does not require programming knowledge, and it easily customized to many other applications, depending on the research questions. Here, we discuss the possible applications and limitations of the system. The low cost and the small learning curve of the RFID-Raspberry Pi system provides a powerful new tool to field biologists.

The NIEHS Superfund Research Program: Epidemiological Research to Address the Complexity of Environmental Exposures and Improve Public Health

The National Institute of Environmental Health Sciences (NIEHS) Superfund Research Program (SRP) funds a wide range of university-based research to address public health concerns arising from hazardous substances in the environment. Since its inception in 1987, the SRP has applied a multidisciplinary approach that combines laboratory-based and population-based studies, as well as community engagement and research translation, to improve our understanding of and minimize the health effects associated with exposures to contaminants. For example, SRP-funded researchers are investigating (1) associations between phthalates and preterm birth, (2) prenatal and early-life exposure to chlorinated compounds and adverse neurological effects in later childhood and adulthood, (3) the links between arsenic in drinking water and various cancers, birth outcomes, and adverse effects on intelligence and motor function in children, and (4) associations between metal mixtures and neurodevelopment. Innovative research from SRP-funded investigators is also addressing challenges in environmental epidemiology through the validation of relevant biomarkers and development of immunoassay-based and field-deployable exposure assessment devices. Moving forward, SRP-funded researchers are focusing on the health effects of complex chemical mixtures, determining relevant chemical exposures, identifying developmental windows of susceptibility, analyzing patterns in epidemiologic and toxicologic data to assess risks to human health presented by hazardous substances, and development of tools to facilitate assessment of exposure and toxicity. This presentation will summarize results from SRP epidemiological studies that have linked hazardous substances to health effects, the impact that these studies have had in human health risk assessment, as well as future projects that will address research priority areas in environmental epidemiology.

Transient isotachophoresis-capillary zone electrophoresis with contactless conductivity and ultraviolet detection for the analysis of paralytic shellfish toxins in mussel samples

The accumulation of paralytic shellfish toxins (PSTs) in contaminated shellfish is a serious health risk making early detection important to improve shellfish safety and biotoxin management. Capillary electrophoresis (CE) has been proven as a high resolution separation technique compatible with miniaturization, making it an attractive choice in the development of portable instrumentation for early, on-site detection of PSTs. In this work, capillary zone electrophoresis (CZE) with capacitively coupled contactless conductivity detector (C4D) and UV detection were examined with counter-flow transient isotachophoresis (tITP) to improve the sensitivity and deal with the high conductivity sample matrix. The high sodium concentration in the sample was used as the leading ion while l-alanine was used as the terminating electrolyte (TE) and background electrolyte (BGE) in which the toxins were separated. Careful optimization of the injected sample volume and duration of the counter-flow resulted in limit of detections (LODs) ranging from 74.2 to 1020ng/mL for tITP-CZE-C4D and 141 to 461ng/mL for tITP-CZE-UV, an 8–97 fold reduction compared to conventional CZE. The LODs were adequate for the analysis of PSTs in shellfish samples close to the regulatory limit. Intra-day and inter-day repeatability values (percentage relative standard deviation, n=3) of tITP-CZE-C4D and tITP-CZE-UV methods for both migration time and peak height were in the range of 0.82–11% and 0.76–10%, respectively. The developed method was applied to the analysis of a contaminated mussel sample and validated against an Association of Official Analytical Chemists (AOAC)-approved method for PSTs analysis by high performance liquid chromatography (HPLC) with fluorescence detection (FLD) after pre-column oxidation of the sample. The method presented has potential for incorporation in to field-deployable devices for the early detection of PSTs on-site.