Standard Method For Quantification Research Articles

A systematic review of the variability of freshly-emitted and aged black carbon based on various measurement techniques.

Black carbon (BC) poses acute negative health and environmental impacts. Accurate BC quantification is important for assessing its impacts and developing effective control strategies. However, the unclear optical properties and numerous methods of BC restrict its accurate quantification. Elemental carbon (EC), refractory black carbon (rBC), and equivalent black carbon (eBC) are terminology used for BC quantification by various techniques, with an essential parameter of mass absorption cross-section (MAC) characterizing its optical properties. This review summarizes the measurement principles, along with the advantages, limitations, and uncertainties of mainstream instruments used for BC quantification, based on findings from numerous studies. Then, we categorically compared the MAC of BC for different emission sources and regions, and those are not consistent. The MAC values of freshly-emitted and aged BC are both influenced by the combustion sources, while the transport process present significant impact on aged BC. Average MAC values of BC for urban, rural, and other areas are 10.1±3.7m2/g, 13.4±5.7m2/g, and 13.7±8.2m2/g at 550nm, respectively. Furthermore, we systematically explored the relationship between eBC, EC, and rBC of freshly-emitted and aged BC to establish unambiguous links between these terms. Averaged mass ratios of eBC/EC, eBC/rBC and EC/rBC are 0.81, 0.94, and 1.32 for freshly-emitted BC, 1.17, 2.14, and 1.60 for aged BC, respectively. Overall, it is urgent to establish standard methods for accurate BC quantification in the future to reduce the uncertainty of quantitative results by different techniques.

Establishment, evaluation and application of a gas chromatography-mass spectrometry method for quantification of volatile organic compounds in ambient air using anisole as an internal standard

Gas chromatography coupled with mass spectrometry (GC-MS) is a standard analytical method for identifying specific volatile organic compounds (VOCs) in urban air samples. Quantification ideally relies on calibration curves using standards for each compound. However, due to the variable composition of VOCs and the limited availability of all standards, this strategy is challenging and not always feasible. Quantification using an internal standard is a simple and accurate approach, as it involves the simultaneous analysis of the internal standard and analytes, eliminating the need for a calibration curve if their concentrations fall within the linear dynamic range. In this study, quantification of VOCs from different chemical classes has been evaluated using a single standard mixture and anisole as an internal standard and compared to the external calibration as standard method for quantification by GC-MS. Both methods showed comparable results, with a relative error within ±30% of the theoretical concentration. Linearity was confirmed in the range from 100 to 4000 μg/L, and the relative standard deviation (RSD) of the relative response factors (RRFs) met the acceptance criteria of ≤30% for each target VOC. Toluene, chlorobenzene, C6-C2 and C6-C3 substituted benzenes yielded similar average RRFs, ranging from 1.280 to 1.601 with an RSD of 7.83%. The methods were applied for quantification of VOCs in ambient air on real samples collected on Radiello adsorbents at one urban location. Using anisole as an internal standard proved to be a simple and reliable method for determining VOC concentrations, offering a convenient alternative to external standard calibration.

Inductively Coupled Plasma Mass Spectrometry Performance for the Measurement of Key Serum Minerals: A Comparative Study With Standard Quantification Methods

ABSTRACTBackgroundInductively coupled plasma mass spectrometry (ICP‐MS) is widely used for the accurate measurement of minerals. However, its application to serum essential mineral measurement has not been fully evaluated. The present study aimed to assess the performance of ICP‐MS for serum minerals by comparing its measurements to those obtained using standard quantification methods.MethodsCross‐sectional data were collected from 282 participants from a single facility in Japan. Serum concentrations of eight key minerals, namely sodium, potassium, calcium, phosphorus, magnesium, iron, zinc, and copper, measured via ICP‐MS and standard methods were compared using Passing–Bablok regression and Bland–Altman plots.ResultsAll minerals, except phosphorus, exhibited good agreement with standard methods, with more stable regression coefficients observed for minerals with greater interindividual variability. After systematically filtering outliers, the mean relative errors were approximately −3% for sodium, potassium, calcium, and magnesium; +5% for iron; 0% for zinc; and −19% for copper. The outliers for iron were primarily due to mild hemolysis, whereas those for zinc were largely attributed to nonhemolysis factors. For phosphorus, the serum total phosphorus concentration measured using ICP‐MS was approximately 3.5 times higher than the serum inorganic phosphorus concentration measured using standard methods, with a weak correlation observed between the two methods.ConclusionThis study provides a practical foundation for future research. Understanding ICP‐MS characteristics will facilitate the development of new approaches in clinical diagnostics.

Development of a Sensitive and Reliable Meso Scale Discovery-Based Electrochemiluminescence Immunoassay to Quantify TDP-43 in Human Biofluids.

Transactive response DNA-binding protein of 43 kDa (TDP-43) is a major component of pathological inclusions in various neurodegenerative disorders, including amyotrophic lateral sclerosis and frontotemporal lobar degeneration. The detection of TDP-43 in biofluids is crucial for the development of diagnostic and prognostic indicators of disease and therapeutic development for TDP-43-related proteinopathies. Despite its potential as a biomarker for numerous neurological disorders, the lack of a sensitive and reproducible TDP-43 assay hinders progress in TDP-43-based therapy development, underscoring the need for an effective and standardized method for accurate quantification. Addressing the limitations of sensitivity and reproducibility in existing assays, in this study, we developed and validated a highly sensitive electrochemiluminescence immunoassay on the Meso Scale Discovery platform. The assay demonstrated the detection of full-length TDP-43 in human biofluids with a limit of detection of 4pg/mL, a working range of 4-20,000 pg/mL, and a total assay time of 16 h. In this study, we developed and validated a sensitive immunoassay for the detection of full-length TDP-43 in human biofluids using the Meso Scale Discovery platform. We used this immunoassay to quantify TDP-43 levels in the plasma and serum of healthy controls and ALS patients. Our results indicate a reduction in full-length TDP-43 in the blood of ALS patients compared to healthy controls.

Design and Optimization of Isothermal Gene Amplification for Generation of High-Gain Oligonucleotide Products by MicroRNAs.

Thermal cycling-based quantitative polymerase chain reaction (qPCR) represents the gold standard method for accurate and sensitive nucleic acid quantification in laboratory settings. However, its reliance on costly thermal cyclers limits the implementation of this technique for rapid point-of-care (POC) diagnostics. To address this, isothermal amplification techniques such as rolling circle amplification (RCA) have been developed, offering a simpler alternative that can operate without the need for sophisticated instrumentation. This study focuses on the development and optimization of toehold-mediated RCA (TRCA), which employs a conformationally switchable dumbbell DNA template for the sensitive and selective detection of cancer-associated miRNAs, specifically miR-21. In addition, we developed variants of hyperbranched TRCA (HTRCA), nicking-assisted TRCA (NTRCA), and hyperbranched NTRCA (HNTRCA) to facilitate exponential amplification by enhancing TRCA through the sequential incorporation of reverse primer (Pr) and nicking endonuclease (nE). By conducting a systematic kinetic analysis of the initial rate and end point signals for varying concentrations of key reaction components, we could identify optimal conditions that markedly enhanced the sensitivity and specificity of the TRCA variants. In particular, HNTRCA, which exploits the synergistic effect of Pr and nE, demonstrated an approximately 3000-fold improvement in the detection limit (260 fM) and a wider dynamic range of more than 4 log orders of magnitude compared to TRCA, thereby evidencing its superior performance. Also, we established a mechanistic model for TRCA that includes the roles of Pr and nE in different amplification processes. Model parameters were fitted to the experimental data, and additional simulations were conducted to compare the four amplification methods. Further tests with real biological samples revealed that this technique showed a good correlation with qPCR in quantifying miR-21 expression in various cell lines (0.9510 of Pearson's r), confirming its potential as a robust and rapid tool for nucleic acid detection. Therefore, the simplicity, high sensitivity, and potential for integration with POC diagnostic platforms make the HNTRCA system suitable for field deployment in resource-limited environments.

Quantitative High-Throughput and High-Sensitivity Glycoprofile Analysis of Therapeutic Glycoproteins Using HILIC-FLD and ESI-MS.

Protein glycosylation is recognized as a Critical Quality Attribute for the biological and therapeutic activity of many recombinant proteins. Therefore, glycosylation should be monitored rigorously to ensure the desired quality, safety, and potency of monoclonal antibodies and other therapeutic glycoproteins. However, glycans are highly heterogeneous structures in proteins, and this poses a challenge for glycoprofile analysis. Hydrophilic interaction liquid chromatography with fluorescence detection has been recognized as a standard method for separation and quantification of released N-glycans after conventional 2-aminobenzamide labeling, but the sample preparation procedure is time-consuming with low sensitivity and poor reproducibility. Here we present a streamlined 96-well plate format platform for high-throughput and high-sensitivity glycan profiling. By taking advantage of rapid glycoprotein denaturation, enzymatic deglycosylation, highly sensitive fluorescent derivation, and on-matrix glycan cleanup, the reaction time has been shortened significantly to approximately 10min. A detailed workflow including two-dimensional chromatography fractionation, exoglycosidase sequential digestion, mass spectroscopy glycan mass confirmation, and data interpretation is described. Potential pitfalls and precautions to be taken to ensure data accuracy are also discussed in this chapter.

The social impacts of circular economy: disclosing epistemological stances and methodological practices

AbstractThe essence of circular economy is about closing the loop of material flows by reducing, reusing, recovering and recycling, ensuring the economic prosperity without any detriment for the environment. However, few information is available about the contribution of circular systems to the social dimension of sustainability (such as well-being, quality of life), how these aspects are evaluated and why they are deemed as important. The purpose of this study is to review those scientific studies that dealt with the social sustainability evaluations of circular systems to highlight new aspects: not only which are the most applied methodologies, which impact categories and indicators are mostly taken into account, but, above all, which is the most diffused epistemological position underlying the evaluation of social aspects of circularity. This last aspect is often disregarded but of utmost importance because it shapes the approaches and procedures choice, and legitimate and justify the insights provided: indeed, they can be the most diverse when dealing with social sciences, which are multiparadigmatic sciences. To the best of our knowledge, no studies have so far emerged that directly investigate the epistemological basis of social impacts within the circular economy. A systematic and hermeneutic review of the scientific literature has been led through a two steps method. The first step, dedicated to search and acquisition, consisted in a standardized replicable process called Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). In a second phase, a hermeneutic review has been conducted on the selected publications. Among the 42 studies reviewed, 7 papers were framed within the post-positivism paradigms, while 83,3% were based on interpretivist paradigms, showing how it is difficult to find consensus on standardized quantification methods for social impacts in circular economy studies. The most recurring qualitative methods relied on personal interpretations, perceptions or observations.

Applying UHPLC-HRAM MS/MS Method to Assess Host Cell Protein Clearance during the Purification Process Development of Therapeutic mAbs.

Host cell proteins (HCPs) are one of the process-related impurities that need to be well characterized and controlled throughout biomanufacturing processes to assure the quality, safety, and efficacy of monoclonal antibodies (mAbs) and other protein-based biopharmaceuticals. Although ELISA remains the gold standard method for quantification of total HCPs, it lacks the specificity and coverage to identify and quantify individual HCPs. As a complementary method to ELISA, the LC-MS/MS method has emerged as a powerful tool to identify and profile individual HCPs during the downstream purification process. In this study, we developed a sensitive, robust, and reproducible analytical flow ultra-high-pressure LC (UHPLC)-high-resolution accurate mass (HRAM) data-dependent MS/MS method for HCP identification and monitoring using an Orbitrap Ascend BioPharma Tribrid mass spectrometer. As a case study, the developed method was applied to an in-house trastuzumab product to assess HCP clearance efficiency of the newly introduced POROS™ Caprylate Mixed-Mode Cation Exchange Chromatography resin (POROS Caprylate mixed-mode resin) by monitoring individual HCP changes between the trastuzumab sample collected from the Protein A pool (purified by Protein A chromatography) and polish pool (purified by Protein A first and then further purified by POROS Caprylate mixed-mode resin). The new method successfully identified the total number of individual HCPs in both samples and quantified the abundance changes in the remaining HCPs in the polish purification sample.

Simultaneous determination of 61 hormones in water by solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry

Concerns over the emergence of steroid hormones as pollutants in water have grown. Steroid hormone compounds present challenges in the simultaneous detection of total residual hormones owing to their analogous structures and diverse types. In this study, we established a rapid and high-throughput continuous online method based on solid phase extraction (SPE) coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of 61 hormone components, including 48 glucocorticoids, 1 mineralocorticoid, 4 androgens, and 8 progesterones, in water. Various SPE columns were investigated to assess their extraction efficiency for enriching and purifying target compounds in a large sample volume (1 L). An HC-C18 SPE column was selected because of its superior performance. Acetonitrile was used as a washing solution during SPE to ensure that the majority of the tested substances achieved recoveries exceeding 70% and effectively avoid interferences from water-soluble components. Various C8 and C18 columns were tested, and the optimal HPLC conditions for hormone retention were established. We systematically evaluated different UPLC columns and mobile phases, including methanol-water and acetonitrile-water systems with 0.1% formic acid added to the aqueous phase. The optimized UPLC conditions were as follows: BEH C18 column (100 mm×2.1 mm, 1.7 μm); column temperature, 40 ℃; flow rate, 0.3 mL/min; injection volume, 5 μL; mobile phase A: 0.1% formic acid aqueous phase; mobile phase B: acetonitrile. Gradient elution was performed as follows: 0-0.5 min, 30%B; 0.5-15.0 min, 30%B-75%B; 15.0-18.0 min, 75%B-98%B; 18.0-19.0 min, 98%B; 19.0-19.1 min, 98%B-30%B; 19.1-20.0 min, 30%B. Both positive- and negative-ion modes were explored in the UPLC-MS/MS experiment to obtain the full scan of the parent ions, and positive mode was finally selected for electrospray ionization (ESI). Two product ions exhibiting strong signals and minimal interference were selected for quantitative and qualitative ion analyses, using an external standard method for quantification. MS/MS was performed in positive-ion (ESI+) mode with multiple reaction monitoring (MRM) scanning. The MS/MS parameters were as follows: atomizing gas pressure, 379 kPa; curtain air pressure, 241 kPa; spray voltage, 5500 V; desolvation temperature, 550 ℃; collision exit voltage (CXP), 13 V; intake voltage (EP), 10 V; and residence time of each ion pair, 0.5 ms. Other instrument settings, such as the collision energy and declustering voltage, were also optimized. The 61 hormones exhibited excellent linear relationships within their corresponding concentration ranges, with correlation coefficients greater than 0.99. The method detection limits (MDLs) were in the range of 0.05-1.50 ng/L. The average recoveries of the 61 hormones across three spiked levels ranged from 62.3% to 125.2%, with relative standard deviations (RSDs, n=6) of 1.1%-10.5%. Using this method, we successfully detected 10 hormone components (cortisone, fluticasone propionate, ciclesonide, betamethasone dipropionate, clobetasone butyrate, diflucortolone valerate, halobetasol propionate, isoflupredone, difluprednate, and hydroxyprogesterone caproate) in various surface water and groundwater samples collected from the Taihu Basin region. The SPE-UPLC-MS/MS method presented in this paper is simple, highly sensitivity, and exceptionally accurate. Thus, it exhibits promising potential for tracing targeted hormone residues in water and will be of great value in monitoring and ensuring water safety. Finally, a regional analysis was conducted on the hormone levels in water, and suggestions were made for the targeted treatment of hormone residues in future sewage treatment processes.

Visualization and standardized quantification of surface charge density for triboelectric materials

Triboelectric nanogenerator (TENG) operates on the principle of utilizing contact electrification and electrostatic induction. However, visualization and standardized quantification of surface charges for triboelectric materials remain challenging. Here, we report a surface charge visualization and standardized quantification method using electrostatic surface potential measured by Kevin probe and the iterative regularization strategy. Moreover, a tuning strategy on surface charge is demonstrated based on the corona discharge with a three-electrode design. The long-term stability and dissipation mechanisms of the injected negative or positive charges demonstrate high dependence on deep carrier traps in triboelectric materials. Typically, we achieved a 70-fold enhancement on the output voltage (~135.7 V) for the identical polytetrafluoroethylene (PTFE) based TENG (neg-PTFE/PTFE or posi-PTFE/PTFE triboelectric pair) with stable surface charge density (5% decay after 140 days). The charged PTFE was demonstrated as a robot e-skins for non-contact perception of object geometrics. This work provides valuable tools for surface charge visualization and quantification, giving a new strategy for a deeper understanding of contact electrification.

Validation of qPCR reference genes in the endangered annual killifish Austrolebias charrua considering different tissues, gender and environmental conditions.

The annual killifish Austrolebias charrua is an endangered species, endemic to the southern region of South America, which inhabits temporary ponds that emerges in the rainy season. The main anthropogenic threat driving the extinction of A. charrua stems from extensive agriculture, primarily due to the widrespread use of glyphosate-based herbicides near their habitats. Annual killifishes have been used as models for ecotoxicological studies but, up to now, there are no studies about reference genes in any Austrolebias species. This represents an obstacle to the use of qPCR-based technologies, the standard method for gene expression quantification. The present study aimed to select and validate potential reference genes for qPCR normalization in the annual killifish Austrolebias charrua considering different tissues, gender and environmental conditions. The candidate reference genes 18 s, actb, gapdh, ef1a, shox, eif3g, and the control gene atp1a1 were evaluated in male and female individuals in three different tissues (brain, liver, and gills) under two experimental conditions (control and acute exposition to Roundup Transorb®). The collected tissues were submitted to RNA extraction, followed by cDNA synthesis, cloning, sequencing, and qPCR. Overall, 18 s was the most stable reference gene, and 18 s and ef1a were the most stable combination. Otherwise, considering all variables, gapdh and shox were the least stable candidate genes. Foremost, suitable reference genes were validated in A. charrua, facilitating accurate mRNA quantification in this species, which might be useful for developing molecular tools of ecotoxicological assessment based on gene expression analysis for environmental monitoring of annual killifish.

Determination of six halogenated solvent residues in olive oil by headspace gas chromatography

The residual amount of halogenated solvents in olive oil is an important indicator of its quality. The National Olive Oil Quality Standard GB/T 23347-2021 states that the residual amount of individual halogenated solvents in olive oil should be ≤0.1 mg/kg and that the total residual amount of halogenated solvents should be ≤0.2 mg/kg. COI/T.20/Doc. No. 8-1990, which was published by the International Olive Council, describes the standard method used for the determination of halogenated solvents in olive oil. Unfortunately, this method is cumbersome, has poor repeatability and low automation, and is unsuitable for the detection and analysis of residual halogenated solvents in large quantities of olive oil. At present, no national standard method for determining residual halogenated solvents in olive oil is available in China. Thus, developing simple, efficient, accurate, and stable methods for the determination of residual halogenated solvents in olive oil is imperative. In this paper, a method based on automatic headspace gas chromatography was established for the determination of residual halogenated solvents, namely, chloroform, carbon tetrachloride, 1,1,1-trichloroethane, dibromochloromethane, tetrachloroethylene, and bromoform, in olive oil. The samples were processed as follows. After mixing, 2.00 g (accurate to 0.01 g) of the olive oil sample was added into a 20 mL headspace injection bottle and immediately sealed for headspace gas chromatography analysis. Blank virgin olive oil was used to prepare a standard working solution and the external standard method for quantification. The solvents used in the preparation of halogenated solvent standard intermediates were investigated and methanol was selected as a replacement for N,N-dimethylacetamide to prepare a halogenated solvent standard intermediate owing to its safety. The effects of different injection times (1, 2, 3, 4, 5, 6 s), equilibration temperatures (60, 70, 80, 90, 100, 110, 120 ℃), and equilibration times (4, 5, 8, 10, 20, 30, 40 min) of the headspace sampler on the detection of the residual amounts of the six halogenated solvents were investigated. The optimal injection time and equilibration temperature were 3 s and 90 ℃, respectively. The method demonstrated good analytical performance for the six halogenated solvents when the equilibration time was 30 min. A methodological study was conducted on the optimized method, and the results showed that the six halogenated solvents exhibited good linear relationships in the range of 0.002-0.200 mg/kg, with correlation coefficients of ≥0.9991. The limits of detection (LODs) and quantification (LOQs) of 1,1,1-trichloroethane and bromoform were 0.0006 and 0.002 mg/kg, respectively. The LODs and LOQs of chloroform, carbon tetrachloride, dibromochloromethane, and tetrachloroethylene were 0.0003 and 0.001 mg/kg, respectively. The average recoveries under different spiked levels were 85.53%-115.93%, and the relative standard deviations (n=6) were 1.11%-8.48%. The established method was used to analyze 13 olive oil samples available in the market. Although no halogenated solvents were detected in these samples, a limited number of samples does not represent all olive oils. Hence, monitoring residual halogenated solvents in olive oil remains necessary for its safe consumption. The LOQs of the method for the six halogenated solvents were significantly lower than that of the COI/T.20/Doc. No. 8-1990 standard method (0.02 mg/kg). In addition, the developed method can be conducted under short operation times with high precision and degree of automation as well as good accuracy. Thus, the proposed method is suitable for the determination and analysis of the residues of the six halogenated solvents in large batches of olive oil samples.

Immunochemistry-based quantification of tumor-infiltrating lymphocytes and immunoscore as prognostic biomarkers in bladder cancer

BackgroundTumor-infiltrating lymphocytes (TILs) and the derived immunoscore (IS) have gained considerable attention over the last decade as prognostic markers in many solid cancers. However, in bladder cancer (BC), their prognostic value is not clearly established.MethodsThe present study aimed to quantify the TILs rates in BC, assess the derived immunoscore, and investigate their prognostic value. An immunochemistry-based quantification of the different subtypes of TILS was performed on paraffin-embedded blocks from patients with invasive urothelial carcinoma of the bladder. We have assessed the rates of TILs, respectively, on peri-tumoral (PT) and intra-tumoral (IT) areas and calculated for each case the corresponding IS which is the index: CD8+/CD3+ TILs. The IS was then classified as low (I0, I1) or high (I2, I3, I4). We included 30 cases in the analysis.ResultsThe median age of patients was 65 years with a sex ratio of 9. TILs densities and distribution were significantly variable between IT and PT areas CD3+ (p = 0.03) and CD8+ (p = 0.004) with the highest rates on the PT areas. In univariate analysis, a low density of CD8+ TILs was significantly associated with an advanced age (p = 0.05), with the presence of lympho-vascular invasion (p = 0.02) and with the absence of specific histological subtype (p = 0.05). A low immunoscore was significantly associated with the presence of lympho-vascular invasion (p = 0.004). No significant association was found between TILs subpopulations, the IS, and the other clinicopathological and survival data. The overall survival (OS) and disease-free survival (DFS) medians were slightly superior in highly T (CD3+/CD8+)-cell infiltrated tumors as well as tumors with a high IS densities. However, the univariate analysis showed that TILs and immunoscore did not impact overall survival (OS) and disease-free survival (DFS).ConclusionTILs and immunoscore might be effective prognostic tools in BC. However, standardized quantification methods and further investigation on larger samples are highly recommended to definitively attest the prognostic value of TILs and IS in BC.

Assessing viability of Escherichia coli in mining-impacted surface waters

Multiple threats to surface water quality, including both fecal pollution and acid mine drainage (AMD) are frequently coincident in mining regions, such as parts of the Appalachian region of the U.S. While it has been established that AMD can have toxic effects to fecal bacteria in surface waters, there is also evidence that AMD may induce a viable but non-culturable (VBNC) state, leaving fecal bacteria undetectable by common standard methods. This study aims to better understand the occurrence of VBNC Escherichia coli in mining-impacted waters (MIW) through three objectives: (1) assess the relationship between MIWs and concentrations of culturable fecal bacteria, (2) compare standard methods for quantification of E. coli in MIW, and (3) investigate whether MIW can induce a VBNC state in E. coli. An analysis of historic data from the Deckers Creek and West Run Watersheds of West Virginia revealed a moderate correlation between pH and E. coli abundance determined via culture and enzyme-based methods. In samples collected from sites within the two watersheds impacted by historic mining activities, the E. coli uidA gene was measured via quantitative polymerase chain reaction (qPCR) and found to be significantly more abundant than E. coli concentrations measured via culture or enzyme substrate coliform test. However, this discrepancy between methods was greatest in samples with a low pH. Further, experiments in which an environmental strain of E. coli was dosed into samples of MIW diluted with saline at different concentrations confirmed that the culturability of E. coli significantly decreased with increasing MIW concentration, while the total number of intact cells, determined via "live/dead” staining and microscopy, remained high. This finding suggests that culture and enzyme methods may underestimate viable E. coli levels in AMD-impacted waters.

Identification of the key aroma compounds in flaxseed milk using stir bar sorptive extraction, aroma recombination, and omission tests

Flaxseed milk is a plant-based dairy alternative that is rich in nutrients. Due to the low concentration of odor compounds in flaxseed milk, it cannot be completely extracted. This poses significant challenges for analysis. Therefore, this study developed a method suitable for extracting volatile compounds from flaxseed milk and compared it with three other extraction methods. It was found that Stir Bar Sorptive Extraction had the best extraction performance, identifying 39 odorants. Flavor dilution factors ranged from 1 to 512, with higher values observed for esters. 13 key odor compounds were identified (odor activity value > 1) using the external standard method for quantification; these included four aldehydes, three pyrazines, two alcohols, two esters, and two other compounds. Pyrazine compounds exhibited the highest concentrations. Aroma recombination and omission experiments showed that nine key odorants contributed significantly to the flavor profile of flaxseed milk, imparting aroma of cucumber, green, mushroom, fruity, sweet, and coconut.

Determination of perchlorate and chlorate in drinks by ultra-performance liquid chromatography-tandem mass spectrometry

To establish a method for determination of perchlorate and chlorate in drinks by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) based on isotopic internal standard method. The perchlorate and chlorate residue in liquid drinks were extracted with methanol, in solid drinks with acetic acid solution, then centrifuged. The supernatant was cleaned-up with PSA/C18 cleanup tube. The separation of perchlorate and chlorate was carried out on a Acquity CSH fluorophenyl column(100 mm×2.1mm, 1.7 μm) and the detection was performed with tandem mass spectrometry with internal standard method for quantification. The peak area ratio of perchlorate and chlorate had a good linear relationship with their mass concentration within their respective linear ranges, with correlation coefficients(r) greater than 0.999. The limits of detection of perchlorate and chlorate were 0.2and 1 μg/L respectively and the limits of quantification were 0.5 and 3 μg/L respectively. The mean recoveries of two compounds were from 84.0% to 105.5% with relative standard deviations from 4.2% to 17.0% and 82.7% to 112.1% with relative standard deviations from 5.5% to 18.4%(n=6), respectively. The perchlorates in 11 kinds of beverage samples were 0.53-4.12 μg/L, chlorates were 3.27-61.86 μg/L. This method is simple, sensitive, accurate and reliable, which is suitable for the determination of perchlorate and chlorate in drinks.

Development and Application of Isotope Labelled Internal Standards in a Sum Parameter Method for Ergot Alkaloid Screening of Food

Ergot alkaloids are a group of toxic compounds, formed by fungi on infested grasses. In 2022, the European Commission set into effect maximum levels for the sum of the twelve major ergot alkaloids in multiple foods. To facilitate the laborious and costly individual quantification of the twelve major ergot alkaloids by HPLC–MS/MS or -FLD, we recently reported a sum parameter method (SPM) for ergot alkaloid quantification. Here, derivatization to lysergic acid hydrazide—a derivative of the mutual ergoline backbone in all ergot alkaloids—allowed simplified determination of all ergot alkaloids in flour via HPLC-FLD. For the measurement of more complex matrices like processed foods, we now developed a MS/MS-based SPM. Two internal standards (IS), isotopically labelled at different positions of the molecule, were synthesized and employed in the MS/MS-measurements. Method performance using either the 13CD3-labelled or the 15N2-labelled IS was evaluated on naturally contaminated rye and wheat flour samples as well as on processed food matrices. Employing the 13CD3-labelled IS leads to lower variances and better consistency with the reference data (obtained by the FLD-based SPM) in flour samples compared to the 15N2-labelled IS. The novel method significantly improves the measurement of ergot alkaloids in complex food matrices, due to their increased selectivity and thus lower interferences. Furthermore, the application of isotope labelled IS obviates the need for time-consuming steps like the determination of recovery rate based, matrix specific correction factors as described in the MS/MS-based European standard method for ergot alkaloid quantification (EN 17425).

Effects of acute physical activity on brain metabolites as measured by magnetic resonance spectroscopy (1H-MRS) in humans: A systematic review

Physical activity (PA) promotes brain health in a variety of domains including cognition, mood, and neuroplasticity. At the neurochemical level, the mechanisms underlying these effects in the brain are not fully understood. With proton Magnetic Resonance Spectroscopy (1H-MRS), it is possible to non-invasively quantify metabolite concentrations, enabling studies to obtain measures of exercise-induced neurochemical changes. This systematic review aimed to examine the existing literature on acute effects of PA on brain metabolites as measured by 1H-MRS. Four databases (Cochrane Central Register of Controlled Trials, PubMed, Embase, and PsycINFO) were searched, identifying 2965 studies, of which 9 met the inclusion criteria. Across studies, Gamma-AminoButyric Acid (GABA) and lactate tended to increase after exercise, while no significant changes in choline were reported. For glutamine/glutamate (Glx), studies were inconclusive. Conclusions were limited by the lack of consensus on 1H-MRS data processing and exercise protocols. To reduce inter-study differences, future studies are recommended to: (1) apply a standardized exercise index, (2) consider the onset time of MRS scans, and (3) follow standardized MRS quantification methods.

Rank classification method for cascade reservoirs considering scale, benefits, and risk consequences

Cascade reservoirs are essential in water resource utilization, flood control, and disaster mitigation. They have been built in several rivers worldwide. The rank classification for cascade reservoirs has significant practical implications for resource distribution decisions and risk management strategies and is also essential for ensuring the security of the basin. Internationally, reservoirs are typically classified based on project scale and dam failure consequences, whereas the risk transmission and superposition effect renders the classification method of reservoirs not directly applicable to cascade reservoirs. Therefore, to address this issue, this study proposed a rank classification method for cascade reservoirs. First, based on the scale, benefits, and risk consequences of dam failure, an index system for the rank classification of cascade reservoirs was established. Second, by constructing social risk criteria for cascade reservoirs and establishing a link among “project rank-reliability index-annual failure probability,” the allowable dam failure losses for different project ranks were determined. Thereafter, considering risk transmission and superposition, the rank classification standard and index quantification method for cascade reservoirs were proposed. Finally, five cascade reservoirs were selected for a feasibility study. The proposed method provides a supplement to the current rank classification standard of water conservancy projects in China and can also serve as a reference for risk assessment and classification management of cascade reservoirs in other countries.

Endometrial Staining of CD56 (Uterine Natural Killer), BCL-6, and CD138 (Plasma Cells) Improve Diagnosis and Clinical Pregnancy Outcomes in Unexplained Infertility and Recurrent IVF Failures: Standardization of Diagnosis with Digital Pathology.

In women with unexplained infertility (UI) and recurrent in vitro fertilization (IVF) failures, the etiology is often unclear. Endometrial immune perturbations and the use of immune markers associated with these dysregulations are of great interest in the diagnosis and treatment of UI. However, reliable biomarkers and standardized quantification methods are lacking. Here, to address endometrial immune dysregulation in UI patients with recurrent IVF failures, we performed endometrial tissue sampling and immunostaining of CD56 (uNK), CD138, and BCL-6. Of these cases, 57.9% had positive CD56 in the endometrial stroma, while 46.1% had positive BCL-6 in the glandular epithelium, and 14.5% of the cases were found to be positive for CD138. Combined staining rates were 60.5%, 68.4%, and 71.05% for (CD56 or BCL-6), (CD56 or CD138), and (CD56, BCL-6, or CD138), respectively. There was a significant correlation between CD56 and BCL-6 positivity, while CD138 positivity was an independent parameter. After the recommended targeted therapy, pregnancy rates were found to increase from 58.5% to 61.6% and 73.8% in CD56-positive, (CD56- or BCL-6-positive), and (CD56-, BCL-6-, or CD138-positive) cases, respectively. Notably, a retrospective evaluation of digital pathology and light microscopy results showed a significant correlation. This study suggests that the examination of CD56, BCL-6, and CD138 in the same endometrial sample may be an effective method in determining the etiology of UI and reaching an early diagnosis and treatment options. Moreover, digital pathology can be used in the evaluation of CD56 and BCL-6 to provide objective, rapid, and reliable results.