Low Concentration Samples Research Articles

Comparison of RT-PCR cycle threshold values between individual and pooled SARS-CoV-2 infected nasopharyngeal swab specimens.

The molecular reverse transcription-polymerase chain reaction (RT-PCR) testing of respiratory tract swabs has become mandatory to confirm the diagnosis of coronavirus disease 2019 (COVID-19). However, RT-PCR tests are expensive, require standardized equipment, and relatively long testing times, and the sample pooling method has been introduced to solve this issue. The aim of this study was to compare the cycle threshold (Ct) values of the individual sample and pooled sample methods to assess how accurate the pooling method was. Repeat RT-PCR examinations were initially performed to confirm the Ct values for each sample before running the pooled test procedure. Sample extraction and amplification were performed in both assays to detect ORF1ab, N, and E genes with a cut-off point value of Ct <38. Overall, there was no difference in Ct values between individual sample and pooled sample groups at all concentrations (p=0.259) and for all pooled sizes. Only pooled size of five could detect the Ct value in the pooled samples for all concentration samples, including low-concentration sample (Ct values 36 to 38). This study highlighted that pooled RT-PCR testing strategy did not reduce the quality of individually measured RT-PCR Ct values. A pool size of five could provide a practical technique to expand the screening capacity of RT-PCR.

Detection of Low-Concentration Biological Samples Based on a QBIC Terahertz Metamaterial Sensor.

Quasi-bound state in the continuum (QBIC) can effectively enhance the interaction of terahertz (THz) wave with matter due to the tunable high-Q property, which has a strong potential application in the detection of low-concentration biological samples in the THz band. In this paper, a novel THz metamaterial sensor with a double-chain-separated resonant cavity structure based on QBIC is designed and fabricated. The process of excitation of the QBIC mode is verified and the structural parameters are optimized after considering the ohmic loss by simulations. The simulated refractive index sensitivity of the sensor is up to 544 GHz/RIU, much higher than those of recently reported THz metamaterial sensors. The sensitivity of the proposed metamaterial sensor is confirmed in an experiment by detecting low-concentration lithium citrate (LC) and bovine serum albumin (BSA) solutions. The limits of detection (LoDs) are obtained to be 0.0025 mg/mL (12 μM) for LC and 0.03125 mg/mL (0.47 μM) for BSA, respectively, both of which excel over most of the reported results in previous studies. These results indicate that the proposed THz metamaterial sensor has excellent sensing performances and can well be applied to the detection of low-concentration biological samples.

Honey, I Shrunk the Columns: Recent Advances in the Design and Use of Narrow Internal Diameter UHPLC Columns

The quest for increased sensitivity for low-concentration samples continues at an accelerated pace, especially in the pharmaceutical and clinical environments. High-end mass spectrometry instruments can enable the analyst to detect analytes at attomolar concentrations, but can be prohibitively expensive and require well-trained scientists to operate and interpret the data. Another approach to improve the sensitivity of high performance liquid chromatography (HPLC) assays is by decreasing the internal diameter (i.d.) of the column. Reducing the column’s internal diameter down from 4.6 mm to 2.1 mm has been in practice for several decades. However, extra caution is required when considering going down below 2.1-mm i.d. into the “capillary-flow” format. This paper describes advances in column design that enable the analyst to achieve high sensitivity and efficiency when reducing the internal diameter of the column. After an introduction outlining the theory of increased sensitivity with narrow and capillary-scale internal diameter columns, a novel 1.5-mm i.d. column format will be presented detailing how sensitivities comparable to capillary-like separations can be achieved with this column geometry. Application examples will demonstrate the performance benefits of utilizing a 1.5-mm i.d. column geometry.

Kinetic study of adsorption and photocatalytic degradation of methylene blue dye using TiO2 nanoparticles with activated carbon

Activated carbon (AC) exhibits limited adsorption capacity for pollutants. Conversely, titanium dioxide (TiO2) demonstrates excellent photocatalytic performance, making it a popular choice for pollutant removal. This study investigates the removal of methylene blue (MB) dye from wastewater using AC, TiO2@AC-2, and TiO2@AC-10 samples via adsorption and photocatalysis. The Energy Dispersive analysis of x-rays (EDAX) has confirmed the presence of Ti, C and O in the prepared samples without any impurities. All the diffraction peaks in x-ray diffractograms indicated the presence of pure anatase TiO2 (tetragonal phase) with no evidence of any other phase. Fourier transform infrared spectroscopy (FTIR) analysis identified a peak around 545 cm−1 in the TiO2@AC-2 sample, indicative of O-Ti-O stretching vibrations. This peak shifted to 602 cm−1 in the TiO2@AC-10 sample. Raman spectroscopy confirmed the presence of carbon (D and G bands) at 1310–1347 cm−1 and 1582–1597 cm−1. Additionally, characteristic Raman active bands for anatase TiO2 were observed at 154 cm−1 (Eg), 204 cm−1 (Eg), 398 cm−1 (B1g), 508 cm−1 (A1g), and 628 cm−1 (Eg). N2 adsorption–desorption isotherms revealed a mesoporous structure for all samples (AC, TiO2@AC-2, and TiO2@AC-10) with hysteresis loops, indicating pores ranging from 2 nm to 50 nm in diameter. Reflectance spectra of TiO2@AC-2 and TiO2@AC-10 displayed absorption edges at 368 nm and 385 nm, respectively, corresponding to a direct band gap of approximately 3.22 eV. Subsequently, these prepared samples were effectively employed for the removal of methylene blue (MB) dye from wastewater utilizing both adsorption and photocatalysis method. Under dark conditions, 20 mg L−1 doses of TiO2@AC-2 and TiO2@AC-10 resulted in 60% and 36% dye adsorption within 60 min respectively. In the presence of UV radiation, the degradation of dye was observed to be 74% and 95% by TiO2@AC-2 and TiO2@AC-10 respectively. This observation indicates that TiO2 nanoparticles along with AC leads to enhanced photocatalytic activity. The Langmuir–Hinshelwood model reveals lower rate constants for AC compared to the composite samples. This is likely because AC lacks inherent catalytic activity, requiring UV light to trigger adsorption. Conversely, TiO2@AC-10 exhibits the highest rate constants (K1 = 24.25 × 10−3 min−1 and K2 = 82.71 × 10−3 min−1), aligning with its higher TiO2 content confirmed by EDAX analysis. This suggests a significantly faster photocatalytic rate and superior degradation efficiency, even at a low sample concentration (20 mg L−1).

A Capillary-Force-Driven, Single-Cell Transfer Method for Studying Rare Cells.

The characterization of individual cells within heterogeneous populations (e.g., rare tumor cells in healthy blood cells) has a great impact on biomedical research. To investigate the properties of these specific cells, such as genetic biomarkers and/or phenotypic characteristics, methods are often developed for isolating rare cells among a large number of background cells before studying their genetic makeup and others. Prior to using real-world samples, these methods are often evaluated and validated by spiking cells of interest (e.g., tumor cells) into a sample matrix (e.g., healthy blood) as model samples. However, spiking tumor cells at extremely low concentrations is challenging in a standard laboratory setting. People often circumvent the problem by diluting a solution of high-concentration cells, but the concentration becomes inaccurate after series dilution due to the fact that a cell suspension solution can be inhomogeneous, especially when the cell concentration is very low. We report on an alternative method for low-cost, accurate, and reproducible low-concentration cell spiking without the use of external pumping systems. By inducing a capillary force from sudden pressure drops, a small portion of the cellular membrane was aspirated into the reservoir tip, allowing for non-destructive single-cell transfer. We investigated the surface membrane tensions induced by cellular aspiration and studied a range of tip/tumor cell diameter combinations, ensuring that our method does not affect cell viability. In addition, we performed single-cell capture and transfer control experiments using human acute lymphoblastic leukemia cells (CCRF-CEM) to develop calibrated data for the general production of low-concentration samples. Finally, we performed affinity-based tumor cell isolation using this method to generate accurate concentrations ranging from 1 to 15 cells/mL.

An open-label feasibility trial of transdermal cannabidiol for hand osteoarthritis

Hand osteoarthritis (OA) is an irreversible degenerative condition causing chronic pain and impaired functionality. Existing treatment options are often inadequate. Cannabidiol (CBD) has demonstrated analgesic and anti-inflammatory effects in preclinical models of arthritis. In this open-label feasibility trial, participants with symptomatically active hand OA applied a novel transdermal CBD gel (4% w/w) three times a day for four weeks to their most painful hand. Changes in daily self-reported pain scores were measured on a 0–10 Numeric Pain Rating Scale (NPRS). Hand functionality was determined via daily grip strength measures using a Bluetooth equipped squeeze ball and self-report questionnaire. Quality of life (QoL) ratings around sleep, anxiety, stiffness and fatigue were also measured. All self-report measures and grip strength data were gathered via smartphone application. Urinalysis was conducted at trial end to determine systemic absorption of CBD. Eighteen participants were consented and 15 completed the trial. Pain ratings were significantly reduced over time from pre-treatment baseline including current pain (− 1.91 ± 0.35, p < 0.0001), average pain (− 1.92 ± 0.35, p < 0.0001) and maximum pain (− 1.97 ± 0.34, p < 0.0001) (data represent mean reduction on a 0–10 NPRS scale ± standard error of the mean (SEM)). A significant increase in grip strength in the treated hand (p < 0.0001) was observed although self-reported functionality did not improve. There were significant (p < 0.005) improvements in three QoL measures: fatigue, stiffness and anxiety. CBD and its metabolites were detected at low concentrations in all urine samples. Measured reductions in pain and increases in grip strength seen during treatment reverted back towards baseline during the washout phase. In summary, pain, grip strength and QoL measures, using smartphone technology, was shown to improve over time following transdermal CBD application suggesting feasibility of this intervention in relieving osteoarthritic hand pain. Proof of efficacy, however, requires further confirmation in a placebo-controlled randomised trial.Trial registration: ANZCTR public trials registry (ACTRN12621001512819, 05/11/2021).

Rapid Concentration and Detection of Bacteria in Milk Using a Microfluidic Surface Acoustic Wave Activated Nanosieve.

Rapid detection of microbes is a key feature for monitoring food quality. Unfortunately, current detection systems rely on labor-intensive and time-consuming lab-based processes that are not suitable for point-of-interest applications and typically require several days before results are available. Here, we demonstrate a microfluidic system capable of rapidly concentrating, fluorescent staining, and detecting bacteria in unprocessed complex biological media such as milk. This concentration is done using a surface acoustic wave-driven microfluidic device which operates based on the Bjerknes force, a force generated on one particle by another in its close proximity. We exploit this effect by exciting a tightly packed bed of 50 μm polystyrene microparticles temporarily with surface acoustic waves within a microfluidic device to capture and release bacterial cells on demand. The bacterial cells are fluorescently stained during capture and then detected using fluorescence microscopy upon release. This device offers a high capturing efficiency (>80%) and a 34 Colony Forming Units (CFU)/mL limit of detection, which is 1 order of magnitude below that of plate counting at 30 CFU per standard 100 μL plate (or 300 CFU/mL). This can be attained in just 1 h of processing at 10 μL/min. With this system, we demonstrate that bacterial detection from extremely low concentration samples down to the order of ∼10 CFU/mL is possible without requiring any additional external pre- or postprocessing.

Mineral composition and heavy metal risk assesment of selected geophagic soils from Tanzania

Geophagy or Pica is the unintentional traditional behavior of eating soil by indigenous people in different countries. practiced in many countries due to nausea among pregnant women and mineral deficiencies without knowing the associated health risks. In this study the mineral composition of geophagic soil and its associated health risk among consumers was determined. Dry soil sticks consumed by women were obtained from open markets in Morogoro, Njombe and Mwanza regions in Tanzania. The elemental concentration of geophagic soil was analyzed using Flame Atomic Absorption spectrophotometer. Health risk assessment methods were used to obtain health information after chronic exposure to geophagic soils. The tests used were Target Hazard Quotients (THQ), Total Target Hazard Quotients (TTHQ) and Cancer Risks (CR). The concentration range of metals in samples obtained from three different regions were 16,335.7–47,773.7 mg/kg for Fe, 46.2–1073.5 mg/kg for Ca, 155.3–514.9 mg/kg for K, 44.5–112.4 mg/kg for Zn, 40.7–95.1 mg/kg for Na, 2.4–66.7 mg/kg for Cu, 109.5–572.6 mg/kg for Mn, 3.8–6.85 mg/kg for Pb, 3.1–93 mg/kg for Ni, 62.7–638.6 mg/kg for Cr and 0.4 mg/kg for Cd. The Provisional Daily Intake (PDI), THQ, TTHQ and CR ranged between 3.0 × 10−3 –34.12 mg/kg/day bw, 0.043–48.75, 34.52–77.36 and 2.55×10−5- 0.23 respectively. The TTHQ>1 was evident for metals in all sampling sites which is indicative of non-carcinogenic health effects. Prolonged exposure to Pb at low concentrations in samples from all the sites can cause pathological effects. The cancer risk values for Pb, Ni, Cr and Cd were <1 in which the consumer is likely not to develop cancer in a life time. Essential minerals – Fe, Ca, Zn, Na, K and toxic metals Pb, Cr, Ni and Cu were detected in all the samples. Cd occurred only in samples from Mwanza region that was below the tolerable daily intake. According to WHO/FAO expert’s joint committee any amount of Pb consumption is not permitted. Given the presence of essential minerals in the geophagic soils which are however accompanied by toxic minerals in some cases which might have carcinogenic effects, prolonged consumption should be discouraged to avoid risks of serious adverse effects to the health of the general population.

Extraction, purification, and clumped isotope analysis of methane (Δ13CDH3 and Δ12CD2H2) from sources and the atmosphere

Abstract. Measurements of the clumped isotope anomalies (Δ13CDH3 and Δ12CD2H2) of methane have shown potential for constraining methane sources and sinks. At Utrecht University, we use the Thermo Scientific Ultra high-resolution isotope-ratio mass spectrometer to measure the clumped isotopic composition of methane emitted from various sources and directly from the atmosphere. We have developed an extraction system with three sections for extracting and purifying methane from high (&gt; 1 %), medium (0.1 % to 1 %), and low-concentration (&lt; 0.1 %) samples, including atmospheric air (∼ 2 ppm = 0.0002 %). Depending on the methane concentration, a quantity of sample gas is processed that delivers 3 ± 1 mL of pure methane, which is the quantity typically needed for one clumped isotope measurement. For atmospheric air with a methane mole fraction of 2 ppm, we currently process up to 1100 L of air. The analysis is performed on pure methane, using a dual-inlet setup. The complete measurement time for all isotope signatures is about 20 h for one sample. The mean internal precision values of sample measurements are 0.3 ± 0.1 ‰ for Δ13CDH3 and 2.4 ± 0.8 ‰ for Δ12CD2H2. The long-term reproducibility, obtained from repeated measurements of a constant target gas, over almost 3 years, is around 0.15 ‰ for Δ13CDH3 and 1.2 ‰ for Δ12CD2H2. The measured clumping anomalies are calibrated via the Δ13CDH3 and Δ12CD2H2 values of the reference CH4 used for the dual-inlet measurements. These were determined through isotope equilibration experiments at temperatures between 50 and 450 °C. We describe in detail the optimized sampling, extraction, purification, and measurement technique followed in our laboratory to measure the clumping anomalies of methane precisely and accurately. This paper highlights the extraction and one of the first global measurements of the clumping anomalies of atmospheric methane.

Impact of introducing a new biological matrix into a validated bioanalytical method: Focus on matrix protein content.

International guidance on bioanalytical method validation recommends the practice of partial validation when introducing a new matrix from the same species into a previously fully validated assay. Planning the partial validation protocol should include an evaluation of analyte chemistry, consideration of sample container materials, and a comparison of properties between the relevant biological matrices. Transition of a serum/plasma-validated bioanalytical method to analysis from a low-protein matrix, such as urine, cerebral spinal fluid, or oral fluid can result in inconsistent analyte recovery. The low recovery can potentially be mistaken for signal suppression or lack of drug stability and may be more pronounced in low-concentration or low-volume samples. In addition, adsorption and absorption interactions with containers may be exacerbated in low-protein matrices. Several possibilities exist for mitigating the impact of non-specific binding and low-protein matrices, including surfactants, bovine serum albumin, and β-cyclodextrin. Finally, higher matrix protein can facilitate analyte stability. Given all this, matrix protein content should not be overlooked when anticipating a partial bioanalytical method validation.

Determination of bisphenols in food and its contact materials migration by magnetic solid-phase extraction coupled with LC-MS/MS

Bisphenols (BPs) pose a pervasive threat to human health due to their potential to leach from plastic packaging into food, thereby endangering the endocrine system. Given their low concentrations in complex samples, the development of a swift, efficient pretreatment method is imperative. In this study, a magnetic covalent organic framework (Fe3O4@TzDa) was synthesized via a layer-by-layer growth approach, utilized in rapid magnetic solid-phase extraction (MSPE) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the determination BPs. This method exhibited remarkable sensitivity, detecting five BPs (BPS, BPF, BPA, BPB, and BPAF) at levels between 0.68 and 0.77 μg L−1. Fe3O4@TzDa nanocomposites maintained an extraction efficiency of approximately 98.0% over five cycles, ensuring robust and repeatable performance. Sample analysis, encompassing plastic bottled water, pear juice, mix juices, pop-top canned curry beef, and box-packed milk, yielded recoveries of 72.0%–114.0% (RSDs <7.5%, n = 5). Furthermore, BPs migration in sports cups and portable bottles was evaluated, revealing migration levels ranging from 0.0014 to 0.0058 mg kg−1. This study advances BP analysis by introducing a reliable method for their efficient determination in beverage samples from plastic food packages. The utilization of Fe3O4@TzDa underscores a pivotal advancement in sample preparation, enhancing both sensitivity and efficiency for the extraction and determination of BPs in food samples.

Solid Phase Extraction Methodology for Robust Isotope Analysis of Atmospheric Ammonium.

The stable nitrogen isotope composition (δ15N) of atmospheric ammonia (NH3) and ammonium (NH4+) has emerged as a potent tool for improving our understanding of the atmospheric burden of reduced nitrogen. However, current chemical oxidation methodologies commonly utilized for characterizing δ15N values of NH4+ samples have been found to lead to low precision for low concentration (i.e., < 5 μmol L-1) samples and often suffer from matrix interferences. Here, we present an analytical methodology to extract and concentrate NH4+ from samples through use of a sample pretreatment step using a solid phase extraction technique involving cation exchange resins. Laboratory control tests indicated that 0.4 g of cation exchange resin (Biorad AG-50W) and 10 mL of 4 M sodium chloride extraction solution enabled the complete capture and removal of NH4+. Using this sample pretreatment methodology, we obtained accurate and precise δ15N values for NH4+ reference materials and an in-house quality control sample at concentrations as low as 1.0 μM. Additionally, the sample pretreatment methodology was evaluated using atmospheric aerosol samples previously measured for δ15N-NH4+ (from Changdao Island, China), which indicated an excellent δ15N-NH4+ match between sample pretreatment and no treatment (y = (0.98 ± 0.05)x + (0.11 ± 0.6), R2 = 0.99). Further, this methodology successfully extracted NH4+ from aerosol samples and separated it from present matrix effects (samples collected from Oahu, Hawaii; pooled standard deviation δ15N-NH4+ = ± 0.5‰,n = 16 paired samples) that without pretreatment originally failed to quantitatively oxidize to nitrite for subsequent δ15N isotope analysis. Thus, we recommend applying this sample pretreatment step for all environmental NH4+ samples to ensure accurate and precise δ15N measurement.

Examination of Analytical Method for Tris(2,3-dibromopropyl)phosphate and Bis(2,3-dibromopropyl)phosphate to Revise the Official Methods Based on "Act on the Control of Household Products Containing Harmful Substances"

In Japan, the use of frame retardants [tris(2,3-dibromopropyl)phosphate: TDBPP and bis(2,3-dibromopropyl)phosphate: BDBPP] in several household textile products is banned under the "Act on the Control of Household Products Containing Harmful Substances." As the official analytical methods for testing these substances have not been revised for over 42 years, several issues such as the using of harmful reagents, have been pointed out. Therefore, we developed a new method to revise the official method in our previous study. In this study, the validity of the developed test method is evaluated at six laboratories using two types of textile samples spiked with TDBPP and BDBPP at three concentrations (4, 8, and 20 µg/g). TDBPP and BDBPP are extracted under reflux using methanol containing hydrochloric acid. TDBPP is analyzed using GC-MS, and BDBPP is also analyzed using GC-MS after methylation with trimethylsilyl diazomethane. Although the accuracy (70-120%), repeatability (<10%), and reproducibility (<15%) of a few samples, mainly low concentration samples, are out of range, overall, the concentration level of detection limits of TDBPP and BDBPP (8 and 10 µg/g) in official analytical methods are quantifiable with sufficient precision using the proposed method. Furthermore, harmful reagents are not used in this method. Thus, the method validated in this study is effective as a revised method for the testing of TDBPP and BDBPP in household textile products.

External quality assessment for molecular detection of Ureaplasma urealyticum in China

Background and aimsA pilot external quality assessment (EQA) scheme for molecular detection of Ureaplasma urealyticum (UU) was conducted by the National Center for Clinical Laboratories (NCCL) to evaluate the testing capabilities of clinical laboratories and the actual performance of DNA-based nucleic acid amplification tests (NAAT) and RNA-based NAATs when applied in clinical settings. Materials and methodsThe EQA panel contained twelve lyophilized samples, including positive samples containing inactivated cell culture supernatants of UU at different concentrations and sterile saline for negative samples. The positive samples were further divided into three groups of high, moderate and low concentrations. The panels were distributed to the participants and the datasets were analyzed according to the qualitative results. ResultsA total of 365 laboratories participated in the EQA scheme, and 360 results submitted by 338 laboratories were collected, of which 96.11 % (346/360) of the returned results and 95.86 % (324/338) of the laboratories were deemed competent. The positive percentage agreement (PPA) was ≥ 97.5 % for high and moderate concentration samples, but varied significantly for low concentration samples, decreasing from 86.94 % to 51.94 % as the sample concentration decreased. Additionally, for low concentration samples, RNA-based NAAT showed higher PPAs than DNA-based NAATs, but these results were specific to UU supernatants used in this study. ConclusionMost of UU detection assays employed by the participants were generally consistent with their estimated limit of detection (LOD), and the majority of participants can reliably detect UU samples with high and moderate concentrations, while the poor analytical performance for low concentration samples requires further improvement and optimization.

Streamlining regular liquid chromatography with MALDI-TOF MS and NMR spectroscopy using automatic full-contact splitless spotting interface and flash-tap fractioning collection

BackgroundHigh-resolution matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and nuclear magnetic resonance (NMR) spectroscopy are powerful tools to identify unknown psychoactive substances. However, in complex matrices, trace levels of unknown substances usually require additional fractionation and concentration. Specialized liquid chromatography systems are necessary for both techniques. The small flow rate of nano LC, typically paired with MALDI-TOF MS, often results in prolonged fractionation times. Conversely, the larger flow rate of semi-preparative LC, used for NMR analysis, can be time-consuming and labor-intensive when concentrating samples. To address these issues, we developed an integrated automatic system that integrated to regular LC. ResultAutomatic spot collector (ASC) and automatic fraction collector (AFC) were present in this study. The ASC utilized in-line matrix mixing, full-contact spotting and real time heating (50 °C), achieving great capacity of 5 μL droplet on MALDI plate, high recovery (76–116%) and rapid evaporation in 2 min. The analytes were concentrated 4–8 times, forming even crystallization, reaching the detection limit at the concentration of 50 μg L−1 for 12 psychoactive substances in urine. The AFC utilizes flexible tubing which flash-tapped the microtube's upper rim (3 mm depth) instead of reaching the bottom. This method prevents sample loss and minimizes the robotic arm's movement, providing a high fractionating speed at 6 s 12 psychoactive compounds were fractionated in a single round analysis (recovery: 81%–114%). Methamphetamine and nitrazepam obtained from drug-laced coffee samples were successful analyzed with photodiode array (PDA) after one AFC round and NMR after five rounds. SignificanceThe ASC device employed real-time heating, in-line matrix mixing, and full-contact spotting to facilitate the samples spotting onto the MALDI target plate, thereby enhancing detection sensitivity in low-concentration and complex samples. The AFC device utilized the novel flash-tapping method to achieve rapid fractionation and high recovery rate. These devices were assembled using commercially available components, making them affordable (400 USD) for most laboratories while still meeting the required performance for advanced commercialized systems.

A pilot study of chlormequat in food and urine from adults in the United States from 2017 to 2023

Chlormequat chloride is a plant growth regulator whose use on grain crops is on the rise in North America. Toxicological studies suggest that exposure to chlormequat can reduce fertility and harm the developing fetus at doses lower than those used by regulatory agencies to set allowable daily intake levels. Here we report, the presence of chlormequat in urine samples collected from people in the U.S., with detection frequencies of 69%, 74%, and 90% for samples collected in 2017, 2018–2022, and 2023, respectively. Chlormequat was detected at low concentrations in samples from 2017 through 2022, with a significant increase in concentrations for samples from 2023. We also observed high detection frequencies of chlormequat in oat-based foods. These findings and chlormequat toxicity data raise concerns about current exposure levels, and warrant more expansive toxicity testing, food monitoring, and epidemiological studies to assess health effects of chlormequat exposures in humans.ImpactThis study reports the detection of chlormequat, an agricultural chemical with developmental and reproductive toxicity, in the U.S. population and U.S. food supplies for the first time. While similar levels of the chemical were found in urine sampled from 2017 to 2022, markedly increased levels were found in samples from 2023. This work highlights the need for more expansive monitoring of chlormequat in U.S. foods and in human specimens, as well as toxicological and epidemiological study on chlormequat, as this chemical is an emerging contaminant with documented evidence of low-dose adverse health effects in animal studies.

Leaching of catalyst platinum from cured silicone elastomers: A preliminary study for comparing reagents

Addition curing systems involve two-part silicones which require the mixture of a silicone polymer with a catalyst to initiate the cure. Platinum is the most commonly used metal catalyst for addition curing of silicones by hydrosilylation which involves the crosslinking by the addition reaction of silicon hydride species to unsaturated bonds, mainly CC, but also CO or CN double bonds. After crosslinking of the polymers, the platinum catalyst cannot be recovered but remains in the silicone materials throughout the entire product life. In the end, platinum is disposed of together with the silicones and is thus lost to the value chain. The overall objective of this work was to develop a recycling process for the recovery of platinum from addition-cured silicone elastomers. In the first step, this was achieved by efficient digestion methods and by optimizing the leaching processes for exemplary commercial silicone elastomer products. Two different silicone materials were investigated, both of which were crosslinked with a platinum catalyst. The initial Pt content in the tested samples was 12.6 ± 0.2 mg/kg for a commercial silicone impression material and 6.3 ± 0.5 mg/kg for a silicone baking mold, measured by graphite furnace atomic absorption spectrometry (GF-AAS). Samples were first frozen with liquid nitrogen to improve brittleness and then crushed with a simple food processor to obtain a silicone granule. Various acid mixtures, mainly based on sulfuric acid, were investigated as digestion methods in order to extract platinum from the silicone network. These had different effects on the dissolution behavior of silicone samples and the amount of platinum extracted in each case. The amount of platinum leached from the filtrate of the digested samples in each case was measured by ICP-OES to evaluate the efficiency of different leaching mixtures. In addition, the dissolved platinum species present in the solutions was identified by UV/VIS as tetrachloridoplatinate(II) complex. The best platinum leaching results so far were obtained with two methods, both of which used a leaching mixture based on sulfuric acid and hexamethyldisiloxane (M2). In the presence of hydrochloric acid, 9.6 ± 1.6 mg platinum/kg was leached from the silicone impression material and 4.2 ± 0.8 mg platinum/kg from the silicone baking mold. With the additional use of aqua regia instead of hydrochloric acid, 10.4 ± 2.8 mg platinum/kg was extracted from the silicone impression material and 4.8 ± 1.0 mg platinum/kg was extracted from the silicone baking mold. These methods were replicated with n = 3. Using statistical evaluation methods (F-test, t-test, and confidence interval), no significant difference was found between these two best methods. Recovery of platinum(0) from leach mixtures has not yet been achieved due to high dilution and very low platinum concentration in samples and will be part of another study.

A novel microfluidic chip-based digital PCR method for enhanced sensitivity in the early diagnosis of colorectal cancer via mSEPT9

BackgroundTo enhance the sensitivity of plasma methylated Septin9 gene (mSEPT9) detection in colorectal cancer (CRC) screening, we developed a microfluidic chip-based digital PCR (dPCR) method suitable for low-concentration samples, aiming to apply it for mSEPT9 detection in CRC diagnosis. MethodsOur microfluidic chip-based dPCR method utilized specific primers and probes with locked nucleic acids (LNAs) modifications for mSEPT9 detection. We evaluated its performance, including detection limit, specificity, and linear range, comparing it with a commercial qPCR reagent kit using the same samples (95 CRC, 23 non-CRC). ResultsThe LNAs-modified dPCR method showed a linear range of 100–104 copies/μL and a detection limit of 100 copies/μL. Clinical testing revealed that our dPCR method exhibited a sensitivity of 82.11 % and specificity of 95.65 % for CRC diagnosis, outperforming the commercial qPCR kit (sensitivity: 58.95 %, specificity: 91.30 %), particularly in Stage I with a diagnostic sensitivity of 90.91 %. Combining mSEPT9 and carcinoembryonic antigen (CEA) improved diagnostic sensitivity to 91.49 %. ConclusionsOur accurate microfluidic chip-based dPCR method, especially in combination with CEA, holds promise for effective CRC screening and timely interventions, offering enhanced mSEPT9 quantification over conventional qPCR.

Development and applications of accelerator mass spectrometry methods for measurement of 14C, 10Be and 26Al in the CENTA laboratory

Terrestrial and extraterrestrial radioisotope research has been strongly dependent on the development of analytical methods which would enable to trace radioisotopes at low concentrations in subgram samples (e.g., in tree rings, ice cores, meteorites, etc.). Accelerator mass spectrometry (AMS) has become the most sensitive technique for ultralow-level analysis of long-lived radioisotopes, such as 14C, 10Be and 26Al. We review developments and applications carried out in the CENTA laboratory, and describe a recently installed fully equipped AMS line, designed for analysis of long-lived radioisotopes from tritium to curium.

Optimization of a pre-concentration method for the analysis of mercury isotopes in low-concentration foliar samples.

Hg isotope analysis in samples from background regions is constrained by the presence of low Hg concentration and therefore requires a pre-concentration method. Existing Hg pre-concentration methods are constrained by long sample processing time and limited sample loading capacity. Using foliar samples as a test case, an optimized Hg pre-concentration method is presented that involves the microwave-assisted digestion of samples for Hg isotope analysis with the addition of a pre-digestion step. Microwave-digested foliar samples and CRMs were transferred to an impinger, reduced with SnCl2, and collected in a 2.25 mL concentrated inverse aqua regia (3:1 HNO3:HCl, v/v). This resulted in an optimal acid concentration in the solution ideal for analysis on MC-ICP-MS. The time for purging with Hg-free N2 was optimized to 30 min and the efficiency of the pre-concentration method was tested using a combination of approaches. Tests performed on pure reagents and matrix of foliar samples spiked with 197Hg radiotracer showed recoveries averaging 99 ± 1.7% and 100 ± 3.0%, respectively. Mercury at concentrations as low as 1.83 ng g-1 was pre-concentrated by digesting aliquots of foliage samples in individual digestion vessels. Recoveries following their pre-concentration averaged 99 ± 6.0%, whereas recoveries of 95 ± 4.7% and 95 ± 2.5% were achieved for NIST SRM 1575a (pine needle) and reagents spiked with NIST SRM 3133, respectively. Analysis using multicollector-ICP-MS showed low fractionation of δ202Hg during sample pre-concentration with no significant mass-independent fractionation. The proposed method is a relatively simple and robust way to prepare Hg samples for Hg isotopic analysis and is suitable even for complex biological matrices.