Sample Matrix Research Articles

Enhanced ICP-MS detection of 24 elemental impurities in complex, high matrix mineral, medicinal lanthanum carbonates according to ICH Q2 (R2).

Validation of the specificity of ICP-MS methods by ICH Q2 (R2) through spiking experiments cannot characterize the multi-atomic interference of complex high matrix samples, possibly affecting the accuracy of the method and reliability of results. In this study, we highlight this issue by investigating the elemental impurities in lanthanum carbonate raw materials and use isotope ratio analysis to overcome this limitation and establishing an accurate ICP-MS method for 24 elemental impurities (listed in ICH Q3D). An Agilent 7900 ICP-MS was used with an automatic sampler for sample determination in the He mode. The isotope ratio of elemental impurities in the spiked samples was analyzed to characterize polyatomic interference and select the exclusive mass number of elemental impurities. While the recovery rate of most elemental impurities met the requirements, that of selenium (default measurement mass number 78Se) exceeded the standard in the matric sample. The isotope ratio of Se (78Se/82Se) was much higher than the theoretical value, indicating that the response intensity of m/z 78 was formed by the combination of 78Se and other mass-to-charge ratios, such as LaOH2+ formed by the large amount of La in the matrix. Therefore, 82 was chosen as the mass number for Se. The validation results indicate that the method has strong specificity, high accuracy, and is simple and facile. This study provides technical support for the control of elemental impurities in lanthanum carbonates and isotope ratios can be used as a supplementary means of spiked recovery rates.

Manufacturing Aluminum-Based Nanocomposites via Stir-Squeeze Casting Combined with Ultrasonication

This study successfully produced aluminum nanocomposites using a stir-squeeze casting process, both with and without ultrasonication (US) assistance. The matrix material utilized was scrap automobile wheel aluminum alloy (A356), with 1% SiC nano particles, averaging a size of 40 nm, serving as the reinforcement material. A comparison was made by also producing A356 aluminum casts with and without the use of US. The produced casts underwent thorough chemical and mechanical characterization, including optical and scanning microscopy, porosity measurement, hardness measurement, compression and tensile testing, as well as wear testing. Additionally, energy-dispersive X-ray spectroscopy (EDS) and X-ray diffraction (XRD) analyses were conducted to assess compositions and confirm the presence of SiC nano particles in the aluminum matrix. Porosity levels were slightly higher in the nanocomposite samples compared to pure matrix samples, attributed to the tendency of pore formation due to improper distribution of ceramic particles, resulting in clustering and agglomeration. However, significant reduction in porosity was observed with the application of ultrasonication, effectively breaking up clusters and agglomerations of reinforcement particles. Regarding mechanical properties, the A356+SiC sample with US exhibited the highest hardness (70.8 HRB), tensile strength (163.25 MPa), and compressive strength (387.2 MPa), along with the lowest abrasive wear loss (0.0017 g) among all types of casts produced in this study.

Matrix Background Screening of an ssDNA Aptamer and Its Identification Against Lactopontin

Lactopontin (LPN) is a highly phosphorylated O-glycosylated acidic protein closely associated with infant gut, brain, and immune development, and its recognition is urgent due to its rising application in fortified dairy products and infant formula. In this study, an ssDNA aptamer against LPN was obtained, among which two kinds of matrix-background-assisted systematic evolution of ligands via exponential enrichment (SELEX) approaches were performed and compared. The direct approach was to utilize the sample matrix as the mixing-incubation background between the ssDNA library and LPN that can theoretically increase screening pressure and simulate practical application scenarios. The indirect approach was to utilize a PBS buffer as a screening background and to include counter-screening steps that adopt the “sample matrix” as a whole as the counter-screening target. Their screening evolutions were monitored through qPCR assays from sequence diversity convergences of each sub-library based on the change in the proportion of hetero- and homo-duplexes from the dissociation curve and melting temperature, which were also verified from the sequence statistics of high-throughput sequencing. The common sequence of Seq.I1II3 from the two approaches was finally fished out as the aptamer through multiple analyses of combining the sequence frequency, secondary structures, homology, and binding assessments, which was demonstrated good specificity and low-nanomolar affinity by qPCR assay (KD, 5.9 nM). In addition, molecular docking and a dynamics simulation were performed for their binding site prediction and affinity confirmation. This study provides a potential identifying element and a basis for accelerating the development of methods for LPN detection in dairy products.

Dairy Chain Safety in the Context of Antibiotic Residues—Current Status of Confirmatory Liquid Chromatography Methods: A Review

With ever-developing analytical capabilities, the protection of the dairy chain from contamination by residues of veterinary drugs is improving. Legislative requirements are an inherent part of this process. Of antibiotics in dairy farming, representatives of the beta-lactams, tetracyclines, sulfonamides, (fluoro)quinolones, aminoglycosides, or polypeptide antibiotics are the most widely used. Due to the typically low levels of antibiotic residues in milk, mass spectrometry is the most commonly used detection technique. However, the interference of the sample matrix is one of its main limiting factors, and therefore, it needs to be eliminated. In the first step, the lipid fraction is removed and proteins are precipitated, followed by solid-phase or liquid–liquid extraction. The current trends include the reduction in the consumption of organic solvents (to reduce occupational hazards and burden to the environment) and automation, eliminating the influence of human error and optimizing the workflow. These trends lead to the development of new microextraction and automated techniques as well as the use of new sorbents and/or (green) solvents of natural origin. To capture the latest developments in the field and the relatively recent aforementioned trends, this review focuses on papers investigating antimicrobial residues in milk that were published between 2015 and 2024, providing an overview of emerging techniques applicable to antibiotic residue detection in milk samples.

Quantitative summary on the human pharmacokinetic properties of cannabidiol to accelerate scientific clinical application of cannabis.

Cannabidiol (CBD) is a non-psychoactive substance that exerts numerous pharmacological benefits, including anti-inflammatory and antioxidant properties. It has received attention as a useful substance for the treatment of intractable pain, seizures, and anxiety, and related clinical trials have continued. However, the CBD pharmacokinetic results between reports are highly variable, making it difficult to clearly identify the pharmacokinetic properties of CBD. The main purpose of this study was to identify CBD clinical pharmacokinetic properties through meta-analysis. In particular, we sought to derive valid, interpretable independent variables and interpret their pharmacokinetic parameter correlations in relation to the large inter-individual and inter-study variability in CBD pharmacokinetics. For this study, CBD-related clinical trial reports were extensively screened and intercomparisons were performed between internal data sets through systematic classification and extraction of pharmacokinetic parameter values. The candidate independent variables associated with interpretation of CBD pharmacokinetic diversity established and explored in this study were as follows: diet, tetrahydrocannabinol (THC) combination, sample matrix type, liver and renal function, exposure route, dosage form, CBD exposure dose, cannabis smoking frequency, multiple exposure. The results of this study showed that CBD pharmacokinetics were influenced (increased plasma exposure by approximately 2-5 times) by diet immediately before or during CBD exposure, and that THC was not expected to have an antagonistic effect on the CBD absorption. The influence of changes in liver function would be significant in CBD pharmacokinetic diversity. Due to decreased liver function, the plasma exposure of CBD increased 2.57-5.15 times compared to healthy adults, and the half-life and clearance showed a 2.58-fold increase and a 5.15-fold decrease, respectively. CBD can be rapidly absorbed into the body (time to reach maximum concentration within 3.18h) by oral, transdermal, and inhalation exposures, and lipid emulsification and nanoformulation of CBD will greatly improve CBD bioavailability (up to approximately 2 times). The pharmacokinetics of CBD generally follow linear kinetic characteristics. The importance of this study is that it suggests key factors that should be considered in terms of pharmacokinetics in further clinical trials and formulations of CBD in the future.

Protein-bridged size-permeable DNAzyme@PEG hollow nanosphere reactor facilitates direct and rapid monitoring of trace circulating microRNAs in undiluted cancer patient serums

The direct monitoring of low levels of circulating microRNAs (miRNAs) in undiluted human serums is of significant clinical importance for the diagnosis of different diseases in the early stages; however, this is highly challenged by both the short lengths, low abundance and homogeneity nature of miRNAs and the complicated sample matrix of human serums. In this work, we show the synthesis of a new protein-bridged and size-permeable DNAzyme@PEG hollow nanosphere reactor (p-DPHNR) that enables direct and rapid detection of trace circulating microRNAs in undiluted serums from prostate carcinoma patients. The hydrophilic PEG shell of p-DPHNR efficiently reduces protein adsorption; its size-permeability only allows short miRNAs to enter the core cavity and protects the core DNA signal probes from digestion; and the confinement of the NIR signal amplification DNA probes in the nanosphere core accelerates the reaction kinetics and minimizes the serum background fluorescence. Such p-DPHNR exhibits a 1.8 pM detection limit with a short 20 min assay time for miRNA-141 and can realize clear discrimination of prostate carcinoma patients from healthy donors, thereby manifesting its robust clinical applications for the convenient and early diagnosis of various diseases

Direct ion chromatographic method for speciation micro analysis of arsenic forms in industrial samples with “rich” matrix composition

The speciation analysis of arsenic has consistently been a subject of great interest. However, it remains challenging to analyze complex matrix samples that contain both arsenic and interfering components. In this case, it can be hard to choose the right combinations of different instrumental methods, or a separation method followed by detection, which is usually done using a spectral approach (hybrid methods). In the production control of copper electrorefining, the determination of the concentration of As (III) and As (V) helps to improve the quality of the cathode copper produced. This work investigated the possibility of directly determining both arsenic forms and total As in an electrolyte bath using ion chromatography (IC) with conductometric detection. The use of the ion chromatographic approach for the determination of As(V) in complex matrix samples such as copper electrolyte must take into account the presence of potential interferences from anions such as sulphates, sulfites, selenites, selenates, etc. The results revealed that the method is accurate and precise, with As(V) quantification limits of 15 µg.L-1 and detection limits of 5 µg.L-1. This method is suitable for assessing various types of arsenic in the production of electrolytic copper, with the aim of replacing the current technique that requires liquid-liquid extraction and ICP-OES detection. This led to the following improvements: Enhanced efficiency: The method eliminates the need for extensive and time-consuming sample preparation for the initial separation of arsenic forms. At the same time, the method's characteristics are comparable to those of ICP-OES with liquid-liquid extraction, which is often used in the speciation analysis of arsenic. The method is environmentally friendly as it avoids the use of organic and poisonous extractants. The method can simultaneously analyze other anions (PO43-, SO42-, F-, Cl-, etc.) with arsenates with appropriate calibration.

Comparative study of virgin and recycled polyethylene terephthalate and polypropylene intermingled thermoplastic composites

AbstractIn this study, a comparative analysis of the use of recycled PET (R‐PET) in synthetic and natural fiber blended composites was made. PET, R‐PET, and PP yarns were folded with reinforcing fibers by commingling technique and then fabrics were prepared using weaving technology. Hot compression molding technology was used for composite fabrication. Tensile and impact characterization revealed the significant influence of reinforcing and TP fibers on mechanical properties. R‐PET matrix glass fiber reinforced composites exhibited 41.77% higher tensile strength than virgin PET and 20.41% higher than PP. In terms of tensile strength for linen reinforced samples, R‐PET shows an impressive 181% improvement over virgin PET and 154% over PP. According to the drop weight test results, glass fiber reinforced R‐PET matrix sample exhibited a peak load value 53% higher than that of the virgin PET matrix, although it was 9.47% lower than the PP matrix. In linen fiber reinforced samples, R‐PET demonstrated a 57.73% improvement over virgin PET but lower by 9.47% compared to the PP matrix. During this time, DSC, TGA, FTIR, and flammability tests were also performed on the composite samples. The results showed that R‐PET has easy processability as a matrix material and provides better wetting of the reinforcing fibers. For this reason, it has been shown that R‐PET matrix samples provide better mechanical properties than both PET and PP matrix samples. It has also been determined that R‐PET is a more suitable matrix material compared with natural fiber reinforced composites.Highlights Commingled yarns were used for preparing composites. R‐PET yarns were used as matrix in preparing glass and linen reinforced composites. R‐PET was compared with virgin PET and PP yarns. R‐PET showed great potential in easy preparation of TP composites. Composites made with R‐PET showed improved thermal and mechanical properties.

A covalent organic framework-derived pretreatment for pesticides in vegetables and fruits

Sample pretreatment is an essential procedure in pesticide analysis, as the matrix effect can significantly influence the results. In this study, a covalent organic framework (COF) was synthesized using 1,2,4,5-tetrakis-(4-formylphenyl)benzene (TFPB) and benzidine (BD) to mitigate the matrix effect in vegetable and fruit samples. This COF was then used to develop a solid-phase extraction (CSPE) method. In addition, the COF was used to create a magnetic COF (MCOF) for use in magnetic solid-phase extraction (MSPE). The synthesized COF and MCOF were thoroughly characterized using scanning electron microscopy (SEM) for morphological analysis, Fourier-transform infrared spectroscopy (FT-IR) for chemical bond identification, and N2 adsorption–desorption measurements for porosity and surface area assessment. Key pretreatment parameters such as buffers, dilution rate, sorbent dosage, extraction time, elution solvent, and reuse number were optimized. The developed CSPE and MSPE showed excellent purification ability for the matrix of vegetable and fruit samples. The reuse test demonstrated that the synthesized COF and MCOF can be reused up to 15 times. Moreover, the developed CSPE and MSPE showed acceptable recoveries in spiked recovery tests, suggesting that these pretreatment methods were feasible for sample purification in pesticide analysis.

Coupling solid phase microextraction to integrated optical sensors with microfluidic open interface

Increasing number of regulations today is very demanding, so many fields need to rely to screening techniques to separate samples whose content need to be confirmed with that field's gold standard approach. Therefore, the development of simple and fast approaches is very much needed. This manuscript presents the development of a microfluidic open interface integrated with ultraviolet-visible (UV–Vis) absorption detection for rapid screening applications. The system was designed as a rapid sensor readout for compounds enriched by solid-phase microextraction devices. Two azo dyes from a liquid water enhancer solution served as model analytes. These two dyes were extracted with the solid phase microextraction fiber and introduced to the system for analysis. Experimental results demonstrated that the system could effectively detect dyes with absorbance at 403 nm, indicating its potential for quantitating analytes with UV–Vis chromophores. Although this sensing method exhibits limited selectivity, it offers a cost-effective, straightforward approach to rapid screening that can be conveniently miniaturized for on-site applications. Further advances could focus on integrating low-cost sensors with specific responses in place of the UV–Vis unit, as well as simplification and miniaturization of the system to improve on-site analysis. Additionally, incorporating autosampler systems could enable high-throughput determinations, broadening the method's applicability. The developed solid phase microextraction method reliably samples and enriches small molecules from complex systems, delivering clean extracts to the selective sensor for readout without interference from the investigated sample matrix.

A practical guide to item bank calibration with multiple matrix sampling

When it is required to estimate item parameters of a large item bank, Multiple Matrix Sampling (MMS) design provides an efficient way while minimizing the test burden on students. The current study exemplifies how to calibrate a large item pool using MMS design for various purposes, such as developing a CAT administration. The purpose of the current study is to explain and provide an example of how to use MMS design for item bank calibration. Two functions of mirt package, mirt() and multipleGroup() were compared using real data. The results of the present study showed that the standard mirt() function is more practical and makes more precise estimations compared to the multipleGroup() function.

Towards harmonization of metal(loid)s determination in conventional and compostable plastics: Comparison of acid digestion protocols in LDPE and PBAT/TPS blends

The determination of metal-containing additives in plastic materials via acid digestion protocols has attracted growing interest to address potential environmental implications. However, the lack of protocol harmonization hinders data comparability within the literature. Here, six acid digestion protocols were employed to determine the metal(loid) content in plastics: these included three different acid mixtures (HNO3 combined with H2SO4, HCl or H2O2) for microwave-assisted digestion, with or without an additional room-temperature digestion step with H2O2.Each protocol was first validated for seven metal(loid)s (As, Cd, Cr, Pb, Sb, Sn and Zn) using a low-density polyethylene (LDPE) certified reference material (ERM®-EC681m). Then, validated protocols were applied on end-use materials, including conventional (i.e., LDPE) and compostable (i.e., PBAT/TPS) plastics.The combination of H2SO4 and HNO3 with a further digestion step with H2O2 was the most suitable protocol: it successfully passed validation thresholds for all metal(loid)s (recoveries in the range 98.6–101.0 %) and yielded the highest concentrations in end-use materials. All other protocols resulted in a less efficient digestion of the sample matrix, leading to lower recoveries and the formation of solid residues. Notably, end-use plastics showed a great variability in metal(loid) concentrations, likely due to their additive-rich composition, in contrast to the minimal content of acid-soluble additives of the reference material.This study represents an initial step towards the harmonization of acid digestion protocols and highlights new challenges in accurately analyzing end-use plastic materials, due to their complex additive composition.

Effect of pre-aging on mechanical reinforcement of the Cu–Fe–Nb composites

Pre-aging treatment was demonstrated to significantly enhance the mechanical properties of Cu–10Fe–1Nb (wt.%) composites. Specifically, after rolling and aging at 400 °C for 8 h, the microhardness, tensile strength, and elongation of the pre-aged samples reached 163 HV, 559 MPa, and 11.2%, respectively. The refinement of Fe-rich phases in pre-aged samples was primarily attributed to the increased shear strain between the Fe-rich phases and the matrix, which was driven by the precipitation-strengthening effect induced by pre-aging. Additionally, the matrix of the pre-aged samples suffered significant refinement as dynamic recovery and recrystallization of the matrix were suppressed during the rolling process, forming a high-density dislocation structure that greatly enhances the work-hardening effect. The main contribution to strength improvement in the Cu–10Fe–1Nb composites was identified as precipitation strengthening, refinement strengthening, and work hardening. Moreover, compared with the sample without pre-aging treatment, the refinement strengthening and work hardening effects of pre-aged samples exhibited substantial improvements, with increment of 49 MPa and 21 MPa, respectively.

Analytical control of residues of the herbicide trifluralin in the assessment of the food safety

Introduction. Trifluralin is a systemic herbicide of the chemical class of dinitroaniline derivatives. During monitoring studies, trifluralin was detected in carrots produced in the Russian Federation. Since trifluralin-based pesticides are not approved for the use in the Russian Federation, residual amounts of the substance are not typical for this crop. To check the contamination of carrots with a pesticide not typical for this crop, confirmatory qualitative and quantitative studies were carried out. Purpose of the work. Increasing the efficiency of identification and reliability of quantitative results when monitoring food products for safety when performing confirmatory analytical studies using the example of determining trifluralin in carrots to assess the safety of food products intended for the consumer. Materials and methods. Gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) was used to identify and quantify trifluralin. Samples were prepared for analysis using the QuEChERS method. Identification was carried out by multiple reaction monitoring (MRM) using a triple quadrupole mass spectrometer using two to four MRM transitions (m/z): 306,1→264,0; 264,0→206,0; 264,0→188,0; 264,0→160,1. Matrix-matched calibration was used for quantitation. Results. The effectiveness of screening studies when monitoring food products using the GC-MS/MS method can be significantly elevated by increasing the number of MRM transitions (at least 3), controlling the reference retention time of the substance, and maintaining the ratio of confirmatory ions. To obtain a reliable quantification of trifluralin content in carrots, the use of a matrix-matched calibration is recommended. Limitation. In the study, only carrots were considered as food products. Conclusion. Confirmation of test results is especially important when determining pesticide residues that are not normally found in a given matrix, or when it is suspected that the maximum permissible level may be exceeded. When conducting a screening study and detecting a food contaminant, contamination of the product can only be reported in advance. Next, a confirmatory analysis is required using a validated quantitative method, including an appropriate calibration procedure. In quantitative analysis, the presence of matrix sample components can cause problems due to sample suppression/enhancement phenomena. Matrix-aware calibration has been shown to be effective in compensating for matrix effects.

Identification of Phospholipids Relevant to Cancer Tissue Using Differential Ion Mobility Spectrometry

Phospholipids are the main building components of cell membranes and are also used for cell signaling and as energy storages. Cancer cells alter their lipid metabolism, which ultimately leads to an increase in phospholipids in cancer tissue. Surgical energy instruments use electrical or vibrational energy to heat tissues, which causes intra- and extracellular water to expand rapidly and degrade cell structures, bursting the cells, which causes the formation of a tissue aerosol or smoke depending on the amount of energy used. This gas phase analyte can then be analyzed via gas analysis methods. Differential mobility spectrometry (DMS) is a method that can be used to differentiate malignant tissue from benign tissues in real time via the analysis of surgical smoke produced by energy instruments. Previously, the DMS identification of cancer tissue was based on a ‘black box method’ by differentiating the 2D dispersion plots of samples. This study sets out to find datapoints from the DMS dispersion plots that represent relevant target molecules. We studied the ability of DMS to differentiate three subclasses of phospholipids (phosphatidylcholine, phosphatidylinositol, and phosphatidylethanolamine) from a control sample using a bovine skeletal muscle matrix with a 5 mg addition of each phospholipid subclass to the sample matrix. We trained binary classifiers using linear discriminant analysis (LDA) and support vector machines (SVM) for sample classification. We were able to identify phosphatidylcholine, -inositol, and -ethanolamine with SVM binary classification accuracies of 91%, 73%, and 66% and with LDA binary classification accuracies of 82%, 74%, and 72%, respectively. Phosphatidylcholine was detected with a reliable classification accuracy, but ion separation setups should be adjusted in future studies to reliably detect other relevant phospholipids such as phosphatidylinositol and phosphatidylethanolamine and improve DMS as a microanalysis method and identify other phospholipids relevant to cancer tissue.

How a mixture of microRNA-29a (miR-29a) and microRNA-144 (miR-144) cancer biomarkers interacts with a graphene quantum dot

MicroRNAs (miRNAs) which are small non-coding RNAs have been reported to be potential cancer biomarker. However, it is difficult to extract such short RNA from a sample matrix. New effective strategies are required. Recently, graphene quantum dots (GQDs) have been used to detect nucleotides in many biosensor platforms, but their applications for miRNA extraction remain limited. GQD was reported to be able to collect short miRNA, but its performance to collect miRNAs with different structure remains unknown. Thus, in this work, the capability of GQD to interact with two different miRNAs is investigated. A mixture of hairpin-like miR-29a and circular miR-144 molecules are used as a representative of two miRNA morphologies. Two systems (a miRNA mixture, comprising 4 of miR-29a and 4 of miR-144, with (miR_GQD) and without GQD (miR)) were studied in comparison. MiRNAs in a mixture (miR) can aggregate, but no permanent miRNA assembly is captured. In contrast, the presence of GQD can rapidly and spontaneously activate the permanent miRNA/GQD clustering. This finding highlights the ability of GQD to be a miRNA collector. Interestingly, all GQD-bound miRNAs do not unfold. This allows the easy accessibility for probes. Also, nano-sized GQD seems to prefer hairpin miR-29a. The free 5’ terminus of miR-29a acts as the sticky end to adhere on GQD. This work highlights the importance of RNA secondary structure on GQD/miRNA aggregation capability. An insight obtained here will be useful for further design of miRNA isolation strategies.

Sequence-Order Frequency Matrix–Sampling and Machine Learning with Smith–Waterman (SOFM–SMSW) for Protein Remote Homology Detection

Sequence-Order Frequency Matrix–Sampling and Machine Learning with Smith–Waterman (SOFM–SMSW) for Protein Remote Homology Detection

A fluorescent platform integrated with a “one-pot” nicking endonuclease signal amplification and magnetic separation for simultaneous detection of tumor markers

Although Enzyme-linked immunosorbent assay (ELISA) has been widely used for biomedical research, simultaneous sensitive and cost-effective detection of multiple biomarkers is challenging. Herein, we proposed a “one-pot” nicking endonuclease signal amplification (NESA)-based fluorescent aptasensor for simultaneous detection of carcinoembryonic antigen (CEA) and alpha fetoprotein (AFP). Firstly, two aptamers were synchronously immobilized on the surface of magnetic nanoparticles (MNPs) by coupling with two complementary DNA (cDNA). CEA and AFP specifically recognized the aptamers and then the released cDNA (ssDNA) from the double-strands (dsDNA) triggers NESA, further breaking two detection probes which were labeled with the fluorescent dye (FAM and ROX) and its quencher (BHQ1 and BHQ2) at the same time. Then, the fluorescence signal of FAM and ROX were restored separately. The results indicated that the fluorescence intensity at the emission wavelength of 518 nm and 610 nm had a positive correlation with CEA and AFP concentrations, respectively. Under the optimum conditions, wider liner range of 1–500 ng mL−1 to CEA and 5–800 ng mL−1 to AFP of this fluorescent aptasensor were successfully obtained, achieving a detection limit of CEA and AFP were 0.7 ng mL−1 and 2 ng mL−1, respectively. Hence, it turned out that the aptasensor strategy can be a promising candidate for developing a newly fluorescence assay for the simultaneous quantitative detection of multiple tumor markers in matrix samples by changing the corresponding sequences of aptamer and fluorescent signal probe, which has great potential for the screening of early cancer.

Is It Just a Game?

Abstract: Building on the long common history of board games and intelligence research, we developed a new deductive reasoning test based on the popular game Mastermind. The research questions of this registered report were: (a) Is a psychometrically sound measurement of the ability to solve Mastermind items possible (i.e., a reliable, uni-dimensional measurement with a good coverage of difficulty)? (b) Is the ability to solve Mastermind items substantially related to other measures of cognitive ability (i.e., matrix test, knowledge test) and need for cognition? (c) Can item difficulty be predicted by the number of colors, positions, premises, and a newly proposed entropy-based index? Based on the results of a pilot study, we developed 30 items and administered them to 351 participants in the preregistered main study using a multiple matrix sampling design. The deductive Mastermind test proved to be (a) a reliable and efficient measure of reasoning across a wide ability range, and (b) showed expectation-consistent patterns for the convergent and divergent measures. (c) The entropy-based index allowed for the prediction of item difficulty to a considerable degree ( R2 = .47). We discuss the ideas of information theory, including entropy, as constructing principles for the rational test development of reasoning tests.

B-229 Extraction-free Nucleic Acid Sample Preparation from Inhibitor Rich Samples via Hyperbaric Heating

Abstract Background Extraction-based nucleic acid sample preparation for PCR is limited by long processing times, liquid transfers, cost, and storage requirements. Extraction-free approaches enable streamlined sample prep but are severely impacted by inhibitors present in biological sample matrices. We previously reported the use of Hyperbaric Heating (HBH) for effective organism lysis and demonstrate here its effectiveness to prepare nucleic acids for PCR from various inhibitor-rich sample matrix types. Methods HBH exposes the sample to high temperature/high pressure environment for a short duration. This process effectively lyses organisms and denatures inhibitors within seconds. Samples are sealed within a pressure-tight metallic vessel, then rapidly heated using magnetic induction. Within the sealed metallic vessel, vaporization of the aqueous sample raises the internal pressure, enabling an internal temperature higher than 100°C. Proprietary sample processing beads containing chelating agents and other proteins protect nucleic acids from fragmentation during heating. Results The efficacy of HBH for PCR amplification was tested against commercial kits and extraction-free methods. HBH treatment of nasal swab samples spiked with inactivated SARS-CoV-2 elicited detection down to 125 viral copies/mL. Viruses such as RSV and Influenza spiked in nasal swab samples demonstrated comparable PCR results as extraction prepped samples. HBH treatment of microbe samples, including Candida albicans spiked in urine and Bacillus subtilis spiked in serum, had comparable PCR threshold cycles as microbe specific DNA extraction methods (Table 1). Conclusions HBH is a generalizable extraction-free nucleic acid sample prep technology affording detection capabilities comparable to commercial sample prep methods. We demonstrated the use of HBH sample prep for detection of several infectious microorganisms in common biological matrices (nasal swab, serum, urine) without additional sample purification. With its ease of use, speed, and lower cost, HBH will be a key tool in applications where nucleic acid amplification is pivotal for diagnostic and research use.