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Analytica Chimica Acta Mar 2022There are several challenges associated with LC-MS/MS bioanalytical method development and validation. Low and variable recovery of some analytes, especially the more...
There are several challenges associated with LC-MS/MS bioanalytical method development and validation. Low and variable recovery of some analytes, especially the more hydrophobic ones, is often challenging. Analytes can be lost to various extents throughout the process of sample collection, storage, before, during, and/or after sample preparation and analysis. The calculation of overall extraction recovery can detect problems of low recovery during sample preparation but does not identify the source(s) of analyte losses. Low overall analyte recovery is the net result of losses that can happen for multiple reasons at all steps of sample preparation and analysis. Therefore, identifying the source(s) of analyte loss during sample preparation can help guide the optimization the bioanalysis conditions to minimize these losses. In this article we propose a practical protocol to systematically identify and quantify the sources of low analyte recovery. This allows the proper choice of strategies to optimize the relevant bioanalytical conditions to minimize analyte losses and improve overall recovery.
Topics: Chromatography, Liquid; Specimen Handling; Tandem Mass Spectrometry
PubMed: 35190119
DOI: 10.1016/j.aca.2022.339512 -
Micromachines Nov 2023The infield measurement of nutrients, heavy metals, and other contaminants in water is still a needed tool in environmental sciences. The Lab-on-a-chip approach can...
The infield measurement of nutrients, heavy metals, and other contaminants in water is still a needed tool in environmental sciences. The Lab-on-a-chip approach can develop deployable instruments that use the standardized analytical assay in a miniaturized manner in the field. This paper presents a Lab-on-a-chip platform for colorimetric measurements that can be deployed for nutrient monitoring in open water (oceans, rivers, lakes, etc.). Nitrite was selected as an analyte. Change to other analytes is possible by changing the reagents and the detection wavelength. In this paper, the principle of the sensor, technical realization, setup of the sensor, and test deployment are described. The sensor prototype was deployed at the Jade Bay (German Bight) for 9 h, measuring the nitrite value every 20 min. Reference samples were taken and processed in the lab. The work presented here shows that an infield measurement using a colorimetric assay is possible by applying Lab-on-a-chip principles.
PubMed: 38004959
DOI: 10.3390/mi14112102 -
Diagnostics (Basel, Switzerland) Sep 2023The usefulness of myositis-specific autoantibodies (MSAs) and myositis-associated autoantibodies (MAAs) for the assessment of idiopathic inflammatory myopathies (IIMs)...
BACKGROUND
The usefulness of myositis-specific autoantibodies (MSAs) and myositis-associated autoantibodies (MAAs) for the assessment of idiopathic inflammatory myopathies (IIMs) is acknowledged, but laboratory standardization remains a challenge. We detected MSAs/MAAs by multi-analytic line immunoassay (LIA) and particle-based multi-analyte technology (PMAT) in a multicenter cohort of patients with IIMs.
METHODS
We tested the sera from 411 patients affected with definite IIM, including 142 polymyositis (PM), 147 dermatomyositis (DM), 19 cancer-associated myositis, and 103 overlap myositis syndrome (OM), and from 269 controls. MSAs/MAAs were determined by 16Ags LIA in all sera, and anti-HMGCR by ELISA in 157/411 IIM sera and 91/269 control sera. The analytical specificity of LIA/HMGCR ELISA was compared with that of PMAT in 89 MSA+ IIM sera.
RESULTS
MSAs/MAAs were positive in 307/411 (75%) IIM patients and 65/269 (24%) controls by LIA (Odds Ratio 9.26, 95% CI 6.43-13.13, < 0.0001). The sensitivity/specificity of individual MSAs/MAAs were: 20%/100% (Jo-1), 3%/99.3% (PL-7), 4%/98.8% (PL-12), 1%/100% (EJ), 0.7%/100% (OJ), 9%/98% (SRP), 5.6%/99.6% (TIF1γ), 4.6%/99.6% (MDA5), 8%/96% (Mi-2), 1.5%/98% (NXP2), 1.7%/100% (SAE1), 4%/92% (Ku), 8.5%/99% (PM/Scl-100), 8%/96% (PM/Scl-75), and 25.5%/79% (Ro52). Anti-HMGCR was found in 8/157 (5%) IIM patients and 0/176 (0%) controls by ELISA ( = 0.007). Concordance between LIA/HMGCR ELISA and PMAT was found in 78/89 (88%) samples. Individual MSAs detected by LIA were associated with IIM subsets: Jo-1 with PM and OM, PL-12 with OM, Mi-2, TIF1γ, and MDA5 with DM, SRP with PM, and PM/Scl-75/100 with OM ( < 0.001 for all).
CONCLUSIONS
Since MSAs are mostly mutually exclusive, multi-specific antibody profiling seems effective for a targeted clinical-serologic approach to the diagnosis of IIMs.
PubMed: 37835823
DOI: 10.3390/diagnostics13193080 -
Biochemia Medica Feb 2023In order to deliver high quality results, detection and elimination of possible analytical interferences, such as lipaemia, is crucial. The aim of this study is to...
INTRODUCTION
In order to deliver high quality results, detection and elimination of possible analytical interferences, such as lipaemia, is crucial. The aim of this study is to evaluate the efficacy of high-speed centrifugation in eliminating lipaemic interference and to define own lipaemic index (LI) for the studied biochemical analytes.
MATERIALS AND METHODS
Evaluated analytes were: albumin, alkaline phosphatase, alanine-aminotransferase (ALT), aspartate-aminotransferase (AST), calcium, creatinine, gamma-glutamyltransferase (GGT), glucose, phosphates, total proteins, urea and total bilirubin. Those analytes and LIs have been analysed in duplicate in the Roche Diagnostics-c8000 analyser in samples centrifuged at 3000 rpm/10 minutes in the SL16 (Thermo Scientific, Waltham, USA) centrifuge and according to an own high-speed centrifugation protocol (12,900 rpm/15 minutes) in the MicroCL17R (Thermo Scientific, Waltham, USA) centrifuge. Lipaemia has been measured in each sample. The efficiency of high-speed centrifugation is verified by the Wilcoxon test (P < 0.05). In cases where significant differences are observed, our own LI is calculated. For ALT and AST, it is verified by McNemar test (P < 0.05. For creatinine, both Wilcoxon and McNemar test were applied.
RESULTS
There were statistically significant differences in analyte concentration before and after high-speed centrifugation for: albumin, creatinine, GGT, glucose, phosphates, urea and total bilirrubin. Own LI is calculated. McNemar test shows statistically significant diferences in the proportion of delivered results before and after high-speed centrifugation in ALT, AST and creatinine.
CONCLUSIONS
This study confirms the efficacy of high-speed centrifugation protocol for all the considered analytes, excepting calcium, alkaline phosphatase and total proteins.
Topics: Humans; Calcium; Creatinine; Alkaline Phosphatase; Centrifugation; Hyperlipidemias; Glucose; Alanine Transaminase; Albumins; Phosphates
PubMed: 36627977
DOI: 10.11613/BM.2023.010703 -
Journal of Extracellular Biology Sep 2023Multi-analyte liquid biopsies represent an emerging opportunity for non-invasive cancer assessment. We developed ONCE (One Aliquot for Circulating Elements), an approach...
Integrating extracellular vesicle and circulating cell-free DNA analysis using a single plasma aliquot improves the detection of HER2 positivity in breast cancer patients.
Multi-analyte liquid biopsies represent an emerging opportunity for non-invasive cancer assessment. We developed ONCE (One Aliquot for Circulating Elements), an approach for the isolation of extracellular vesicles (EV) and cell-free DNA (cfDNA) from a single aliquot of blood. We assessed ONCE performance to classify HER2-positive early-stage breast cancer (BrCa) patients by combining EV-associated RNA (EV-RNA) and cfDNA signals on = 64 healthy donors (HD) and non-metastatic BrCa patients. Specifically, we isolated EV-enriched samples by a charge-based (CB) method and investigated EV-RNA and cfDNA by next-generation sequencing (NGS) and by digital droplet PCR (ddPCR). Sequencing of cfDNA and EV-RNA from HER2- and HER2+ patients demonstrated concordance with in situ molecular analyses of matched tissues. Combined analysis of the two circulating analytes by ddPCR showed increased sensitivity in ERBB2/HER2 detection compared to single nucleic acid components. Multi-analyte liquid biopsy prediction performance was comparable to tissue-based sequencing results from TCGA. Also, imaging flow cytometry analysis revealed HER2 protein on the surface of EV isolated from the HER2+ BrCa plasma, thus corroborating the potential relevance of studying EV as companion analyte to cfDNA. This data confirms the relevance of combining cfDNA and EV-RNA for HER2 cancer assessment and supports ONCE as a valuable tool for multi-analytes liquid biopsies' clinical implementation.
PubMed: 38046436
DOI: 10.1002/jex2.108 -
Molecules (Basel, Switzerland) Nov 2020Nuclear Magnetic Resonance (NMR) spectroscopy is a quantitative analytical tool commonly utilized for metabolomics analysis. Quantitative NMR (qNMR) is a field of NMR... (Review)
Review
Nuclear Magnetic Resonance (NMR) spectroscopy is a quantitative analytical tool commonly utilized for metabolomics analysis. Quantitative NMR (qNMR) is a field of NMR spectroscopy dedicated to the measurement of analytes through signal intensity and its linear relationship with analyte concentration. Metabolomics-based NMR exploits this quantitative relationship to identify and measure biomarkers within complex biological samples such as serum, plasma, and urine. In this review of quantitative NMR-based metabolomics, the advancements and limitations of current techniques for metabolite quantification will be evaluated as well as the applications of qNMR in biomedical metabolomics. While qNMR is limited by sensitivity and dynamic range, the simple method development, minimal sample derivatization, and the simultaneous qualitative and quantitative information provide a unique landscape for biomedical metabolomics, which is not available to other techniques. Furthermore, the non-destructive nature of NMR-based metabolomics allows for multidimensional analysis of biomarkers that facilitates unambiguous assignment and quantification of metabolites in complex biofluids.
Topics: Animals; Biomarkers; Humans; Metabolome; Metabolomics; Nuclear Magnetic Resonance, Biomolecular
PubMed: 33158172
DOI: 10.3390/molecules25215128 -
Plants (Basel, Switzerland) Aug 2023In many analytical chemical procedures, organic solvents are required to favour a better global yield upon the separation, extraction, or isolation of the target... (Review)
Review
In many analytical chemical procedures, organic solvents are required to favour a better global yield upon the separation, extraction, or isolation of the target phytochemical analyte. The selection of extraction solvents is generally based on the solubility difference between target analytes and the undesired matrix components, as well as the overall extraction procedure cost and safety. Hansen Solubility Parameters are typically used for this purpose. They are based on the product of three coordinated forces (hydrogen bonds, dispersion, and dipolar forces) calculated for any substance to predict the miscibility of a compound in a pure solvent, in a mixture of solvents, or in non-solvent compounds, saving time and costs on method development based on a scientific understanding of chemical composition and intermolecular interactions. This review summarises how Hansen Solubility Parameters have been incorporated into the classical and emerging (or greener) extraction techniques of phytochemicals as an alternative to trial-and-error approaches, avoiding impractical experimental conditions and resulting in, for example, saving resources and avoiding unnecessary solvent wasting.
PubMed: 37631219
DOI: 10.3390/plants12163008 -
Cell May 2023Phenotypic sex-based differences exist for many complex traits. In other cases, phenotypes may be similar, but underlying biology may vary. Thus, sex-aware genetic... (Review)
Review
Phenotypic sex-based differences exist for many complex traits. In other cases, phenotypes may be similar, but underlying biology may vary. Thus, sex-aware genetic analyses are becoming increasingly important for understanding the mechanisms driving these differences. To this end, we provide a guide outlining the current best practices for testing various models of sex-dependent genetic effects in complex traits and disease conditions, noting that this is an evolving field. Insights from sex-aware analyses will not only teach us about the biology of complex traits but also aid in achieving the goals of precision medicine and health equity for all.
Topics: Animals; Female; Male; Models, Genetic; Multifactorial Inheritance; Phenotype; Quality Control; Sex Characteristics; Genome-Wide Association Study; Guidelines as Topic; Gene-Environment Interaction; Humans
PubMed: 37172561
DOI: 10.1016/j.cell.2023.04.014 -
Molecules (Basel, Switzerland) Apr 2023Infrared spectroscopy (wavelengths ranging from 750-25,000 nm) offers a rapid means of assessing the chemical composition of a wide range of sample types, both for... (Review)
Review
Infrared spectroscopy (wavelengths ranging from 750-25,000 nm) offers a rapid means of assessing the chemical composition of a wide range of sample types, both for qualitative and quantitative analyses. Its use in the food industry has increased significantly over the past five decades and it is now an accepted analytical technique for the routine analysis of certain analytes. Furthermore, it is commonly used for routine screening and quality control purposes in numerous industry settings, albeit not typically for the analysis of bioactive compounds. Using the Scopus database, a systematic search of literature of the five years between 2016 and 2020 identified 45 studies using near-infrared and 17 studies using mid-infrared spectroscopy for the quantification of bioactive compounds in food products. The most common bioactive compounds assessed were polyphenols, anthocyanins, carotenoids and ascorbic acid. Numerous factors affect the accuracy of the developed model, including the analyte class and concentration, matrix type, instrument geometry, wavelength selection and spectral processing/pre-processing methods. Additionally, only a few studies were validated on independently sourced samples. Nevertheless, the results demonstrate some promise of infrared spectroscopy for the rapid estimation of a wide range of bioactive compounds in food matrices.
Topics: Spectroscopy, Near-Infrared; Anthocyanins; Spectrophotometry, Infrared; Food; Polyphenols
PubMed: 37049978
DOI: 10.3390/molecules28073215 -
Beilstein Journal of Nanotechnology 2023Lateral flow assays (LFAs) are currently the most widely used point-of-care testing technique with remarkable advantages such as simple operation, rapid analysis,... (Review)
Review
Lateral flow assays (LFAs) are currently the most widely used point-of-care testing technique with remarkable advantages such as simple operation, rapid analysis, portability, and low cost. Traditionally, gold nanoparticles are employed as tracer element in LFAs due to their strong localised surface plasmon resonance. However, this conventional LFA technique based on colorimetric analysis is neither useful to determine critical analytes with desired sensitivity, nor can it quantify the analytes. Various signal amplification strategies have been proposed to improve the sensitivity and the quantitative determination of analytes using LFAs. One of the promising strategies is to enhance the photothermal properties of nanomaterials to generate heat after light irradiation, followed by a temperature measurement to detect and quantify the analyte concentration. Recently, it has been observed that the nanoscale architecture of materials, including size, shape, and nanoscale composition, plays a significant role in enhancing the photothermal properties of nanomaterials. In this review, we discuss the nanoarchitectonics of nanomaterials regarding enhanced photothermal properties and their application in LFAs. Initially, we discuss various important photothermal materials and their classification along with their working principle. Then, we highlight important aspects of the nanoscale architecture (i.e., size, shape, and composition) to enable maximum light-to-heat conversion efficiency. Finally, we discuss some of the recent advances in photothermal LFAs and their application in detecting analytes.
PubMed: 37822722
DOI: 10.3762/bjnano.14.82