With recent evolution of cannabis legalization around the world and multiplication of cannabis derived products, identifying and qualifying cannabis consumption has a proven interest. Although blood, plasma, and urine are common matrices widely used in toxicology laboratories, oral fluid presents specific advantages. In the context of doping tests, addiction consultation or roadside checks, where other matrices are impractical to collect or can be adulterated, oral fluid is a promising matrix that allows a non-invasive, rapid, and monitored self-sampling. However, available devices required a consequent volume of oral fluid, more than 250 µL, sometimes difficult to collect. We present here a fully optimized quantitative method for seven cannabinoids, including four metabolites, in oral fluid, Δ9-tetrahydrocannabinol, 11-hydroxy-Δ9-tetrahydrocannabinol and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol; cannabidiol, 7-hydroxy and 7-carboxycannabidiol; and cannabinol. After self-collection of 20 µL using an accurate and precise volumetric absorptive microsampling device (VAMS), cannabinoids were derivatized with 2-fluoro-1-methylpyridinium p-toluenesulfonate to increase sensitivity. The successive steps of the proposed method, including biosampling, 1 h sample preparation with derivatization, and acquisition by ultrahigh-performance liquid chromatography coupled to high-resolution mass spectrometry, were fully optimized. A limit of quantification of 0.5 ng/mL (≈10 pg per sampling) was thus targeted, adapted to the legal threshold required by the authorities and to clinical monitoring. Applied to six cannabis consumers, the proposed method made it possible to quantify in 20 µL oral fluid samples, Δ9-tetrahydrocannabinol ranging from 0.5 to 6236 ng/mL, cannabidiol from 0.6 to 190 ng/mL and cannabinol from 0.5 to 118 ng/mL.

Interest in bile acids (BAs) is growing due to their emerging role as signaling molecules and their association with various diseases such as colon cancer and metabolic syndrome. Analyzing BAs requires chromatographic separation of isomers, often with long run times, which hinders BA analysis in large studies. Here, we present a high-throughput method based on liquid chromatography-tandem mass spectrometry to quantify BAs in mouse samples. After acidic protein precipitation in the presence of a comprehensive mixture of stable isotope-labeled internal standards (SIL-ISs), BAs are separated on a biphenyl column by gradient elution at basic pH. Quantification is performed using a six-point calibration curve. Except for the separation of β- and ω-muricholic acid (MCA) species, a rapid separation of 27 BA species was achieved in a run time of 6.5 min. Plasma quality controls (QCs) were used to evaluate intra- and inter-day precision. The CV was less than 10% for most BA species and exceeded 20% only for glycohyodeoxycholic (GHDCA) and taurohyodeoxycholic acid (THDCA) due to the lack of a corresponding SIL-IS. The limit of quantification (LoQ) was tested using diluted QCs and was found to be compromised for some BA species as a result of insufficient isotopic purity of the SIL-IS, leading to significant interference with the respective analyte. Finally, we tested the mouse sample material requirements for plasma, bile, and liver samples and determined BA concentrations in C57/BL6N wild-type mice. In conclusion, the LC-MS/MS method presented here permits a rapid and reproducible quantification of the major murine BAs.

There is growing interest in road pollution that enters surface waters. Additive chemicals used in the manufacture of plastics, including tyre rubber, are mobile pollutants that pose a threat to aquatic life. Therefore, an ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed to measure 25 plastic additive chemicals in road runoff and water of retention ponds used to manage road runoff. A straightforward direct injection methodology was adopted to minimise sample handling and risk of contamination. Using this approach, the method quantitation limits (MQLs) ranged from 4.3 × 10 to 13 µg/L. These were adequate to determine most chemicals at or below their freshwater predicted no-effect concentration (PNEC). Method trueness ranged from 18 to 148% with most chemicals being within 80-120%. The method was applied to water from four retention ponds in series to measure additive chemicals entering the ponds (i.e., in road runoff) and passing through each pond. Greatest concentrations were observed in road runoff during heavy rainfall following dry weather. Here, 1,3-diphenylguanidine (DPG) exceeded its current PNEC of 1.05 µg/L. Notably, N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine-quinone (6PPD-quinone) was determined at 0.13 µg/L which is greater than its lowest acute toxicity threshold (0.095 µg/L). Similarity in additive chemical concentrations throughout the retention ponds during steady flow suggests little or no removal occurs. However, further studies are needed to assess the fate and removal of plastic additive chemicals in retention ponds and the risk posed to aquatic environments. Such research can be facilitated by this newly developed UHPLC-MS/MS method.

Metal ion pollution poses a global concern due to its significant risks to both human health and environmental well-being. The toxicity of these ions can increase when they coexist, interacting with each other and with other harmful substances, even at low concentrations. Therefore, an accurate, rapid, and cost-effective methodology is urgently needed for the simultaneous quantification of multiple metal ions. This study presents a new approach for the multiplexed detection of various metal ions (Ag, Cu, Hg, Al, Pb, Fe, Fe, Zn, Ni, Cd, and Ca) using a triple-emission nanoprobe comprising carbon dots and distinctly capped CdTe quantum dots, specifically green-emitting glutathione -quantum dots and red-emitting 3-mercaptopropionic acid-quantum dots. The method achieved high accuracy by analysing first- and second-order photoluminescence data with distinct advanced chemometric tools. R values for partial least squares and unfolded partial least square models exceeding 0.9 for several metal ions at low concentrations (mmol L) were obtained. Additionally, PL second-order data yielded significantly better results than PL first-order data, attributed to the distinct behaviour of the metal ions over time. Interestingly, it was also noted for the first time the significant contribution of the molar ratio between the metal ions on the models' accuracy. This novel method provides a highly accurate and efficient way to detect multiple metal ions simultaneously, paving the way for improved environmental monitoring and pollution assessment. The utilization of the proposed method contributes to a better understanding of the complex interactions in mixed metal ion systems, allowing for earlier detection and mitigation of metal ion contamination threats.

Synthetic esters are widely applied in aviation turbine engines due to its excellent lubrication performance. Mixed acids with varying chain lengths and degrees of isomerization are often used in the esterification process to ensure the comprehensive lubricity of synthetic esters. In industrial production, the feeding ratio of mixed acids is usually adjusted by viscosity properties of synthetic esters, so the exact proportion of each acid in the final product is unknown. A method has been developed here to rapidly extract fatty acids used in commercially available synthetic ester lubricants through transesterification reactions, with methanol as the ester exchange reagent. Accurate qualitative and quantitative analysis of it is performed using GC-MS. Compared with the saponification extraction technique, this method is fast, simple, and easy to operate. The formation of this analytical approach can not only simplify the identification of fatty acids in industrial synthetic esters, but also help to determine the accurate ratio of mixed acids in laboratory esterification products, thereby providing technical support for the upgrading and replacement of products.

The precise target recognition and nuclease-mediated effective signal amplification capacities of CRISPR-Cas systems have attracted considerable research interest within the biosensing field. Guided by insights into their structural and biochemical mechanisms, researchers have endeavored to engineer the key biocomponents of CRISPR-Cas systems with stimulus-responsive functionalities. By the incorporation of protein/nucleic acid engineering techniques, a variety of conditional CRISPR-Cas systems whose activities depend on the presence of target triggers have been established for the efficient detection of diverse types of non-nucleic acid analytes. In this review, we summarized recent research progress in engineering Cas proteins, guide RNA, and substrate nucleic acids to possess target analyte-responsive abilities for diverse biosensing applications. Furthermore, we also discussed the challenges and future possibilities of the stimulus-responsive CRISPR-Cas systems in versatile biosensing.

This study investigated the molecular mechanisms of the interactions between three antitumor active monosaccharides and human serum albumin (HSA) using spectroscopic and electrochemical analyses, supplemented by molecular docking simulations. The antitumor efficacy of these monosaccharides can be significantly enhanced by covalent drug coupling, while HSA, with its long half-life and low immunogenicity, provides new opportunities for the development of advanced antitumor drug delivery systems. The results showed that these monosaccharides effectively burst the fluorescence of HSA. Thermodynamic analysis revealed that Fucose undergoes a spontaneous, exothermic process that decreases entropy, while the binding of Mannose and Galactose is entropy-driven. Notably, the addition of these three monosaccharides slightly compacts the structure of HSA, stabilizing its transport and delivery in vivo, with the binding strength categorized as Fucose > Mannose > Galactose. These variations in binding constants explain the differences in efficacy and potential side effects in antitumor therapy. Further studies have shown that the interaction between monosaccharides and HSA improves drug stability and targeting, thereby enhancing antitumor activity. An in-depth study of these interactions may provide new insights into the design and optimization of antitumor drugs and the further development of novel antitumor therapies.

The analysis of polar analytes with the help of hydrophilic interaction liquid chromatography (HILIC) using classic methods of high-performance liquid chromatography is not without its downsides. In these applications, acetonitrile is prevalent as main eluent and sample diluent. This results not only in slow diffusion processes during the separation, but also in often unstable sample solutions where polar analytes are concerned. Furthermore, there are ecological concerns. With the use of supercritical fluid chromatography (SFC) which uses supercritical carbon dioxide as eluent, and other green solvents as alternative for the sample preparation, the separation of polar analytes could be vastly improved with this technique. Fast diffusion within carbon dioxide led to shorter analysis times and higher plate numbers. Regarding sample diluents, small alcohols such as ethanol and 2-propanol, as well as acetone, yielded promising results while analytes showed higher solubility and stability within these solvents compared to acetonitrile. Other green solvents such as dihydrolevoglucosenone (Cyrene) and dimethyl carbonate were found to be unsuitable sample diluents for applications in SFC.

Sulfite, widely used as a food additive, performs indispensable functions in the field of food sterilization, bleaching, and antisepsis. However, the overuse of sulfite may destroy food nutrition and pose health risks to people. In this work, an innovative BODIPY-based fluorescent probe (BODIPY-DBC) was constructed for highly sensitive recognition of sulfite. The BODIPY-DBC probe possessed both colorimetric and ratiometric dual mode, a low detection limit (33.12 nM), high sensitivity, a wide pH usage range (5-12), a fast response time (2 min), and superior fluorescence imaging capability for detecting sulfite. The recognition mechanism was certificated by H NMR titration, HRMS analysis, and DFT calculation. The BODIPY-DBC probe was not only loaded on test strips for detecting sulfite conveniently with the naked eye, but also employed to detect sulfite content in real food samples to ensure food safety. Furthermore, it also achieved excellent performances for monitoring sulfite in dual-channel fluorescence imaging (HeLa cell and zebrafish).

Quorum sensing is a physiological phenomenon of microbial cell-to-cell information exchange, which relies on the quorum sensing signal molecules (QSSMs) to communicate and coordinate collective processes. Quorum sensing enables bacteria to alter their behavior as the population density and species composition of the bacterial community change. Effective detection of QSSMs is paramount for regulating microbial community behavior. However, traditional detection methods face the shortcomings of complex operation, high costs, and lack of portability. By combining the advantage of biosensing and nanomaterials, the biosensors play a pivotal significance in QSSM detection. In this review, we first briefly describe the QSSM classification and common detection techniques. Then, we provide a comprehensive summary of research progress in biosensor-based QSSM detection according to the transduction mechanism. Finally, challenges and development trends of biosensors for QSSM detection are discussed. We believe it offers valuable insights into this burgeoning research area.

The Brazilian National Metrology Institute produced a suite of certified reference materials (CRMs) intended as internal standards (ISs) for quantitative nuclear magnetic resonance (qNMR). Being a ratio primary method, the use of qNMR in organic chemistry has already crossed the borders of research laboratories, despite the cost of instrumentation. The International Bureau of Weights and Measures (BIPM) proposed eight potential qNMR ISs. Four candidate materials were selected for their solubility in various solvents and distinct chemical shifts, making them suitable for qNMR analysis of diverse analytes. The certification process compared orthogonal primary methods such as mass balance, qNMR, freezing-point depression, and coulometry, to ensure independent value assignment. Different approaches were compared to assess batch homogeneity and stability. While directly comparing the main compound's chromatographic area proved to be a quick and fit-for-purpose approach, the determination of individual impurities provided lower uncertainties but required more laborious work. CRM batches of maleic acid, dimethyl sulfone, potassium hydrogen phthalate, and dimethyl terephthalate were delivered with over 999.8 mg g purity and uncertainty in the range of 0.6 to 3 mg g (k = 2). The literature shows certification procedures for qNMR ISs whose traceability chain is exclusively based on qNMR measurements. As opposed to that, the methodology presented here provides robust certified values assigned by methods independent of qNMR, in accordance with BIPM recommendations and less prone to qNMR biases. The CRMs developed in this work have already been used for SI-traceable purity evaluation of compounds such as drugs and pesticides, by laboratories in Brazil and abroad.

In glycomics, two data repositories, GlycoPOST and UniCarb-DR, have been developed to accumulate experimental data generated by glycomics and glycoproteomics mass spectrometry experiments. In order to enhance the interrelation between these two data repositories, we have upgraded the framework for both of them; we have unified their respective data submission systems and constructed a mechanism that can automatically cross-reference corresponding entries. In addition to this integration, the metadata registration system was also extended so that liquid chromatography experiments can be reported according to standard reporting guidelines specified by MIRAGE (Minimum Information Required for A Glycomics Experiment). Furthermore, by augmenting the visualization software used in UniCarb-DR, we have been able to introduce new functionality into GlycoPOST to enable the visualization of unpublished experimental identification result files during an embargo period defined by the data provider. As a result, this work introduces a new framework by which glycomics researchers can take advantage of GlycoPOST and UniCarb-DR in an integrated manner.

The scourge of drug addiction and abuse poses a significant challenge to society. Opioid drugs acting on μ-opioid receptor (OPRM1) make it one of the pivotal targets for drug addiction. In the past decade, sewage analysis has become a prevalent method of drug monitoring. However, traditional methods of detecting drugs in sewage are cumbersome, and rapid detection methods are relatively lacking. To address this, an innovative OPRM1-SNAP-tag/CMC method to directly identify drug components in sewage was established. Cell membrane chromatography (CMC) is an affinity chromatography technique which effectively detects receptor affinity substances. Cells constructed with high expression of specific receptor could be used to screen for compounds acting on the receptor. CMC based on OPRM1 provides a potentially convenient and effective tool for the detection of targeted drug components in sewage. In this study, the selectivity, reproducibility, column lifetime, and carryover of the CMC column had been assessed. Initially, we eluted the collected domestic sewage with methanol and acetonitrile, and the retention peaks were observed on the CMC system. Subsequently, without any preliminary sample preparation, we directly injected filtered samples of suspicious sewage into the OPRM1-SNAP-tag/CMC system, where we observed retention peaks as well. The retained components were further identified as morphine by using UPLC-MS/MS. In conclusion, the OPRM1-SNAP-tag/CMC method stands out as a reliable and robust model for the detection of drug components in sewage. It provides a valuable analytical tool for frontline drug control efforts, enhancing our capacity to monitor and mitigate the impact of drug abuse on society.

Raman spectroscopy is an extensively explored vibrational spectroscopic technique to analyze the biochemical composition and molecular structure of samples, which is often assumed to be non-destructive when carefully using proper laser power and exposure time. However, the inherently weak Raman signal and concurrent fluorescence interference often lead to Raman measurements with a low signal-to-noise ratio (SNR), especially for biological samples. Great efforts have been made to develop experimental approaches and/or numerical algorithms to improve the SNR. In this study, we proposed an ensemble learning approach to recover and denoise Raman measurements with a low SNR. The proposed ensemble learning approach was evaluated on 986 pairs of Raman measurements, each pair of which consists of a low SNR Raman spectrum and a high SNR reference Raman spectrum from the exact same fungal sample but uses 200 times the integration time. Compared with conventional methods, the Raman measurements recovered by the proposed ensemble learning approach are more identical to high SNR reference Raman measurements, with an average RMSE and MAE of only 1.337 × 10 and 1.066 × 10, respectively; thus, the proposed ensemble learning approach is expected to be a powerful tool for numerically improving the SNR of Raman measurements and further benefits rapid Raman acquisition from biological samples.

Epidermal growth factor receptor (EGFR) mutations play a key role in the development of a variety of cancers. Rapid detection and screening of EGFR mutation types in patients are of great significance for early treatment of patients. In this study, a highly sensitive fluorescent biosensing system based on lanthanide ion-doped multi-type upconversion nanoparticles (UCNPs) combined with polymerization reaction signal amplification was designed and constructed for the simultaneous detection of L858R and 19Del mutations. Two upconversion nanoparticles (NaYF:Yb, Er and NaYF:Yb, Tm) with unique upconversion fluorescence profiles were first prepared using Er and Tm as activators, respectively. Subsequently, the UCNPs were enriched by cDNA complementary hybridization and atom transfer radical polymerization (ATRP) reactions to enhance the signal. Next, the tDNA/cDNA hybrids were cleaved using specific restriction endonucleases to detach UCNPs aggregates from the surface of the magnetic beads. Finally, the fluorescence signal in the supernatant was detected after magnetic separation. The simultaneous quantitative detection of the two EGFR mutations was achieved by analyzing the changes in signal intensity of the characteristic upconversion fluorescence spectra of the two encoded UCNPs at their respective emission peaks. The detection range of the method was from 10 fM to 10 nM, and the detection limits were 2.44 fM for L858R and 2.13 fM for 19Del. The sensing system was able to effectively differentiate between wild-type and other mutation types, and its detection results were consistent with qPCR. The excellent performance of the sensor suggests its promising application in the diagnosis and precision treatment of NSCLC.

Many chemicals in food packaging can leach as complex mixtures to food, potentially including substances hazardous to consumer health. Detecting and identifying all of the leachable chemicals are impractical with current analytical instrumentation and data processing methods. Therefore, our work aims to expand the analytical toolset for prioritizing and identifying chemical hazards in food packaging. We used a high-performance thin-layer chromatography (HPTLC)-based bioassay to detect genotoxic fractions in paperboard packaging. These fractions were then processed with non-targeted liquid chromatography high-resolution mass spectrometry (LC-HRMS/MS) and machine learning-based toxicity prediction (MLinvitroTox). The HPTLC bioassay detected four genotoxic zones in extracts of the paperboard. One-dimensional HPTLC separation and targeted fraction collection reduced the number of chemical features extracted from paperboard and detected with LC-HRMS by at least 98% (from 1695-2693 to 14-50). The entire process was successful for spiked genotoxic chemicals, which were correctly prioritized in the fractionation and non-target analysis workflow. The native chemical with the strongest genotoxicity signal was identified with a suspect list as 5-chloro-2-methyl-4-isothiazolin-3-one and confirmed with LC-HRMS/MS and HPTLC bioassay. Toward identification of the remaining unknown genotoxicants, two-dimensional HPTLC further reduced the number of chemical features. Genotoxicity predictions with MLinvitroTox based on molecular fingerprints of the unknown signals derived from their MS2 fragmentation spectra helped prioritize two chemical features and suggested candidate structures. This work demonstrates strategies for using HPTLC, HRMS, and toxicity prediction to help identify toxicants in food packaging.

Natural flavonoids have been shown to have many pharmacological activities. Efficient and continuous enrichment of total flavonoids with high content and low mobile phase usage from complex natural products is greatly needed at the moment. In this study, a new continuous chromatography system (CCS) with 6 zones and the 18-column dynamic tandem connection technique was developed and used to enrich total flavonoids from Epimedium koreanum Nakai (EKN). The 18 columns were divided into 6 zones, and the principle of a dynamic series of three columns was adopted for each zone to achieve continuous automatic separation and enrichment of total flavonoids under the control of a logic control valve. The CCS separation conditions were established based on single-column chromatography and a theoretical calculation model of the CCS. By means of the self-designed device and method, 485.11±3.16 g of total flavonoids were isolated from 16.2 kg of EKN. It is worth noting that the total content of 18 types of flavonoids in the samples enriched by the CCS was increased from 2.84±0.07% to 88.29±0.22%, the total recovery rate was 92.20±0.38%, and the RSD of each flavonoid was less than 5.0%. Furthermore, compared with single-column chromatography filled with the same volume of chromatography filler, the entire process saved about 2/3 of the mobile phase usage. In summary, the developed device and method could efficiently and continuously enrich total flavonoids from EKN with high-content and low mobile phase usage and would have a wide application prospect in the separation and enrichment of natural products.

Sensitive and accurate detection of protein biomarkers is crucial for disease diagnosis, especially for early diagnosis. Here, we describe surface-enhanced Raman scattering (SERS)-based CRISPR/Cas12a assays (S-CRISPR) for protein biomarker detection. Firstly, an S-CRISPR-driven enzyme-linked immunosorbent assay (S-CasLISA) was developed utilizing a capture antibody coated on a microplate to recognize the target and a detection antibody labeled with active DNA to trigger the activity of CRISPR/Cas12a. With this assay, we achieved detection of prostate-specific antigen (PSA) as models at the picogram level. The limit of detection (LoD) of S-CasLISA was 0.17 pg mL and in the range of 0.1 pg mL to 10 ng mL. Further, we applied aptamer to S-CRISPR (S-Apt-CRISPR), combining the high sensitivity of SERS with the high selectivity of aptamers, while simplifying the operation process of CRISPR detection of protein biomarkers. The proposed S-Apt-CRISPR also could detect picogram-level PSA and without repeated washing steps. The LoD of S-Apt-CRISPR was 0.35 pg mL and in the range of 0.1 pg mL to 10 ng mL. Both SERS-based CRISPR/Cas12a assays were validated with clinical samples and demonstrated accuracy consistent with the chemiluminescence immunoassay. The introduction of the CRISPR/Cas12a system with SERS has the effect of improving the analytical capabilities of the system, thereby broadening and facilitating its application in the analysis of sensitive and accurate protein biomarkers.

In this work, we explore a new dispersive liquid-liquid microextraction (DLLME) method to selectively separate chemical species of Cd and Zn in saline waters. It is based on the use of the magnetic ionic liquid (MIL) methyltrioctylammonium tetrachloroferrate ([N][FeCl]), which allows an efficient and environmentally friendly extraction of the target species. In addition, the paramagnetic component in the MIL simplifies the separation step required in DLLME, allowing for fast separation and recovery of the extracted species with a magnet, without a centrifugation step. The optimum conditions for the separation by MIL-DLLME were 3.3 mg mL MIL, sample pH = 8, and an extraction time of 30 min. Under these conditions, metal chlorocomplexes (99.7% and 81.0% of total metal concentration for Cd and Zn, respectively) were quantitatively separated, remaining the free cations in the aqueous samples. In a second step, the extracted metal species were back-extracted with 1 mol L HNO and a re-extraction time of 15 min. For cadmium, this acidic solution separated the neutral complex CdCl (60.5%), while CdCl (21.5%) and CdCl (18.1%) remained in the organic phase. For Zn, the anionic complex ZnCl (17.3%) was retained by the organic reagent, while ZnCl (45.7%) and ZnCl (37.0%) were re-extracted by the nitric acid solution. The separation of the chemical species of metals along the three liquid phases used allowed their quantification in several samples of real seawater and a certified reference material.

Mass spectrometry imaging (MSI) is a promising analytical method to visualize the distribution of lipids in biological tissues. To clarify the relationship between cellular distribution and lipid types in a tissue, it is crucial to achieve both an improvement in ion detection sensitivity and a reduction in the ionization area. We report methods for improving the efficiency of ion transfer to a mass spectrometer and miniaturizing the extraction area of a sample for tapping-mode scanning probe electrospray ionization (t-SPESI), atmospheric pressure sampling, and ionization methods. To verify the efficacy of the new t-SPESI measurement system, MSI was performed on mouse testes with a pixel size of 5 µm. Lipid images of the testes from wild-type (WT) and lysophospholipid acyltransferase 3 (LPLAT3) knockout mice revealed the characteristic distribution of docosahexaenoic acid-containing phospholipids (DHA-PLs). A comparison of the ion images obtained by MSI and optical images of the same tissues stained with hematoxylin and eosin suggested that the distribution of DHA-PLs was significantly altered by spermatogenesis in the WT mouse testes.

Magnetic ionic liquids (MILs) have proven effective as capture reagents for foodborne bacterial pathogens; however, there are currently no published studies regarding their use with foodborne, non-enveloped viruses. In this study, a protocol was evaluated for capture and recovery of bacteriophage MS2, a human norovirus surrogate, and purified viral genomic single stranded RNA (ssRNA) from an aqueous suspension using MILs. Transition metal-based MILs showed similar capture and recovery efficiency for both targets. A rare earth metal-based MIL showed much greater capture efficiency than the transition metal-based MILs, but displayed similar recovery. All tested MILs showed slightly higher capture and recovery efficiency for free RNA in comparison to intact virus, though overall trends were similar, and most MILs could recover both targets at as little as 10 PFU/mL intact MS2 or copies/mL purified RNA. A plaque assay confirmed that contact with MILs did not significantly reduce viral infectivity. Adjusting MIL volume gave no significant changes in capture or recovery, likely due to interplay between volume for the hydrophobic MIL and dispersion. Reducing the elution volume gave a slight increase in recovery, indicating MILs could be used for target enrichment after further optimization. MILs could also capture MS2 from romaine lettuce rinsate at comparable or even higher levels than from pure suspension, though loss in recovery was observed when the rinsate was prepared in an alkaline elution buffer. Overall, these results demonstrate the potential utility of MILs as concentration reagents for foodborne viruses, particularly for in-field applications.