Interlaboratory Comparison Research Articles

Successful Detection of Delta and Omicron Variants of SARS-CoV-2 by Veterinary Diagnostic Laboratory Participants in an Interlaboratory Comparison Exercise.

Throughout the COVID-19 pandemic, veterinary diagnostic laboratories have tested diagnostic samples for SARS-CoV-2 both in animals and over 6 million human samples. An evaluation of the performance of those laboratories is needed using blinded test samples to ensure that laboratories report reliable data to the public. This interlaboratory comparison exercise (ILC3) builds on 2 prior exercises to assess whether veterinary diagnostic laboratories can detect Delta and Omicron variants spiked in canine nasal matrix or viral transport medium. The ILC organizer was an independent laboratory that prepared inactivated Delta variant at levels of 25 to 1000 copies per 50 µL of nasal matrix for blinded analysis. Omicron variant at 1000 copies per 50 µL of transport medium was also included. Feline infectious peritonitis virus (FIPV) RNA was used as a confounder for specificity assessment. Fourteen test samples were prepared for each participant. Participants used their routine diagnostic procedures for RNA extraction and real-time reverse transcriptase-PCR. Results were analyzed according to International Organization for Standardization (ISO) 16140-2:2016. Overall, laboratories demonstrated 93% detection for Delta and 97% for Omicron at 1000 copies per 50 µL. Specificity was 97% for blank samples and 100% for blank samples with FIPV. No differences in Cycle Threshold (Ct) values were significant for samples with the same virus levels between N1 and N2 markers, nor between the 2 variants. The results indicated that all ILC3 participants were able to detect both Delta and Omicron variants. The canine nasal matrix did not significantly affect SARS-CoV-2 detection.

Zinc Isotope Measurement by MC‐ICP‐MS in Geological Certified Reference Materials

Here we present an analytical method for Zinc (Zn) isotopic measurements using “standard‐sample bracketing” on a Nu Sapphire multi‐collector inductively coupled plasma‐mass spectrometer. The effects of Zn mass fraction and HNO3 molarity mismatch between the standard and the sample, as well as the presence of matrix elements have been evaluated. Long‐term reproducibility of better than ±0.03‰ for δ66Zn (2s) was routinely obtained. Accurate measurements were achieved when CNi/CZn < 0.001, CTi/CZn < 0.03, CBa/CZn < 0.05, CNa/CZn and CAl/CZn < 0.5, CMg/CZn < 0.1, CFe/CZn < 5 and CCu/CZn as well as CCd/CZn < 10. High‐precision Zn isotopic determination were performed on twenty‐one widely available geological certified reference materials, with an overall range of ~ 0.62‰ (0.15 to 1.07‰), which is nearly twenty times the current analytical precision (0.03), and the results are in agreement with most previously published data within 2s. Among them, δ66ZnJMC 3‐0749L of eleven geological certified reference materials are reported for the first time: 0.22 ± 0.03‰ (dolerite, DR‐N), 0.23 ± 0.02‰ (gabbro, GSR‐10), 0.20 ± 0.04‰ (microgabbro, PM‐S), 0.15 ± 0.02‰ (andesite, GSR‐2), 0.22 ± 0.08‰ (diorite, GSR‐9), 0.19 ± 0.07‰ (syenite, GSR‐7), 0.30 ± 0.06‰ (granite, GSR‐1), 0.25 ± 0.00‰ (rhyolite, GSR‐11), 0.37 ± 0.01‰ (shale, GSR‐5), 0.21 ± 0.03‰ (limestone, GSR‐6) and 0.20 ± 0.03‰ (hornblendite, GSR‐15). The novel Zn isotopic data from these certified reference materials can be used for future interlaboratory comparisons.

Transferability and reproducibility of exposed air-liquid interface co-culture lung models

BackgroundThe establishment of reliable and robust in vitro models for hazard assessment, a prerequisite for moving away from animal testing, requires the evaluation of model transferability and reproducibility. Lung models that can be exposed via the air, by means of an air-liquid interface (ALI) are promising in vitro models for evaluating the safety of nanomaterials (NMs) after inhalation exposure. We performed an inter-laboratory comparison study to evaluate the transferability and reproducibility of a lung model consisting of the human bronchial cell line Calu-3 as a monoculture and, to increase the physiologic relevance of the model, also as a co-culture with macrophages (either derived from the THP-1 monocyte cell line or from human blood monocytes). The lung model was exposed to NMs using the VITROCELL® Cloud12 system at physiologically relevant dose levels. ResultsOverall, the results of the 7 participating laboratories are quite similar. After exposing Calu-3 alone and Calu-3 co-cultures with macrophages, no effects of lipopolysaccharide (LPS), quartz (DQ12) or titanium dioxide (TiO2) NM-105 particles on the cell viability and barrier integrity were detected. LPS exposure induced moderate cytokine release in the Calu-3 monoculture, albeit not statistically significant in most labs. In the co-culture models, most laboratories showed that LPS can significantly induce cytokine release (IL-6, IL-8 and TNF-α). The exposure to quartz and TiO2 particles did not induce a statistically significant increase in cytokine release in both cell models probably due to our relatively low deposited doses, which were inspired by in vivo dose levels. The intra- and inter-laboratory comparison study indicated acceptable interlaboratory variation for cell viability/toxicity (WST-1, LDH) and transepithelial electrical resistance, and relatively high inter-laboratory variation for cytokine production. ConclusionThe transferability and reproducibility of a lung co-culture model and its exposure to aerosolized particles at the ALI were evaluated and recommendations were provided for performing inter-laboratory comparison studies. Although the results are promising, optimizations of the lung model (including more sensitive read-outs) and/or selection of higher deposited doses are needed to enhance its predictive value before it may be taken further towards a possible OECD guideline.

A New Method for Dead Time Calibration and a New Expression for Correction of WDS Intensities for Microanalysis.

Observed photon count rates must be corrected for detector dead time effects for accurate quantification, especially at high count rates. We present the "constant k-ratio" method, a new approach for calibrating dead time for wavelength dispersive spectrometers by measuring k-ratios as a function of beam current. The method is based on the observation that for a given emission line at a specific take-off angle and electron beam energy, the intensity ratio from two materials containing the element should remain constant as a function of beam current, if the dead time calibration is accurate. The method has the advantage that it does not rely on the linearity of the beam current picoammeter, yet also allows the analyst to evaluate the picoammeter linearity, another critical parameter in EPMA calibration. By simultaneously comparing k-ratios for all spectrometers, one can also ascertain k-ratio consensus, essential for inter-laboratory comparisons. We also introduce improved dead time expressions and provide best practices on how to perform these instrument calibrations using this new "constant k-ratio" method. These improvements enable quantitative analysis of major and minor elements with high accuracy at high beam currents, simultaneously with trace elements with high sensitivity, for point analyses and X-ray mapping.

THE STUDY OF METROLOGICAL CHARACTERISTICS OF THE OBTAINED SUBSTANCES OF FAT-SOLUBLE VITAMINS BY INTERLABORATORY COMPARISON TESTS

Relevance. This article discusses the possibility of using substances isolated from natural raw materials of fat-soluble vitamins retinol-palmitate (vitamin A), cholecalciferol (vitamin D3), tocopherol-acetet (vitamin E) in order to create domestic standard samples of approved type (SSS). The use of SSS in analytical practice, as well as in scientific research, makes it possible to ensure the accuracy and reliability of experimental results, to ensure the traceability of the result when performing analyses within the framework of repeatability and reproducibility (precision). The paper addresses issues related to the production and standardization of domestic pharmaceutical substances, as well as issues of standardization and conformity assessment of finished products of the pharmaceutical industry and other sectors of the national economy (food industry, agriculture, perfumery and cosmetics, the industry of production of goods for children's practice, etc.). The purpose of the work. To establish the quantitative content of fat-soluble vitamins A, D3, E, in the obtained substances claiming the status of SS with the approved type, as well as to determine their stability and uniformity by means of interlaboratory comparison tests (hereinafter referred to as ICT). Material and methods. The quantitative content of fat-soluble vitamins in the obtained substances was assessed by HPLC in 6 accredited analytical laboratories. The results were processed by evaluating the data provided for the presence of outliers and quasi-outliers using the Kohren, Grubbs and Mandel method. Results. The conducted studies under the comparison program showed that the results of the quantitative determination of the obtained vitamins are within the norms of the certified characteristic "Mass fraction of vitamin, %" in relation to the content of the determined characteristic in reference substances of imported production. Conclusions. As a result of the statistical data obtained, we have proved that the substances obtained from fat-soluble vitamins A, E, D3 comply with regulatory requirements based on the results of interlaboratory comparisons and can be recommended for the manufacture of domestic SS with the approval of their type, and it can also be assumed that the manufacturing processes of domestic and imported SS are harmonized. The quality of domestic SS is at an identical level with respect to foreign SS, but each is produced exclusively in the legal field of the manufacturer's state and is legitimate in its territory of application, which once again confirms the high need to create a domestic reference pharmaceutical base.

109 Inter- and Intra-Laboratory Comparison of 6 Dustiness Testing Methods: Towards the Development of an OECD Testing Guideline

Abstract Dustiness is a key parameter describing the ability of powder materials to generate dust during agitation. The main goal of this work was to generate and assess the scientific basis for the validation and applicability of 6 dustiness methods to nanomaterials, and development of a subsequent OECD testing guideline. Six dustiness methods (rotating drum, small rotating drum, continuous drop, vortex shaker, fluidizer and venturi) were subjected to an intra- and inter-laboratory comparison (ILC) in which 15 international laboratories participated. ILC tests were conducted for 6 materials (3 TiO2 and 3 SiO2) of different chemical natures and dustiness levels. Each participating laboratory conducted at least 3 replicates per material. Harmonization of procedures, methods and data treatment took place prior to testing. Results from the ILC were assessed considering the different reported metrics such as respirable mass and particle number dustiness index. Overall, the intralaboratory variability for the different methods and metrics was under 30%. Variation between laboratories was generally higher for respirable mass than particle number dustiness index. These variations were mostly attributed to differences in setups such as tubing length or instrumentation. All methods except the venturi, which was characteristic for presenting low differences between materials and opposed classification to the rest of the methods, showed relatively similar material ranking. For all methods and laboratories the calculated z-score (measure of the deviation of each laboratory from the true value) was <2 as based on ISO 13528, which indicates gratifying results. Funding: EU H2020 Research and Innovation Programme under Grant Agreement 814401.

97 Transferability and Reproducibility of Exposed Air-Liquid Interface co-Culture Lung Models

Abstract The establishment of reliable and robust in vitro models for hazard assessment, moving away from animal testing, requires evaluation of model transferability and reproducibility. Lung models that can be exposed via the air, by means of an air-liquid interface are promising in vitro models for evaluating the safety of nanomaterials (NMs) after inhalation exposure. We performed an inter-laboratory comparison study to evaluate the transferability and reproducibility of a lung model consisting of the human bronchial cell line Calu-3 as monoculture and, to increase the relevance of the model, also as co-culture with macrophages (derived from the THP-1 monocyte cell line or from human blood monocytes). Exposure employed the VITROCELL® Cloud12 system at physiologically relevant doses. Overall, the results of the seven participating laboratories are quite similar. No effects of lipopolysaccharide (LPS), quartz (DQ12) or titanium dioxide (TiO2) NM-105 exposure on cell viability and barrier integrity were seen in the monoculture and co-culture models. Most laboratories showed that LPS induced cytokine release in the co-culture models, but not the monoculture model. Exposure to DQ12 or TiO2 did not increase cytokine release, probably due to too low deposited doses. Interlaboratory variability indicated acceptable variation for cell viability measurements and relatively high variation for cytokine measurements. Based on the transferability and reproducibility of the model and its air exposure we are confident that, based on detailed SOPs, performing a new round of interlaboratory comparison, and increasing the deposited doses of NMs, these models may be taken further towards a possible OECD guideline.

Application of KU812 cells for assessing complement activation related effects by nano(bio)materials

Immunocompatibility issues related to nano(bio)materials, particularly liposomal formulations, involving activation of the complement system have been relatively well described however, they highlight the importance of preclinical evaluation of such interactions. These complement-mediated hypersensitivity reactions, in which basophils are implicated, are associated with complement activation-related pseudoallergy (CARPA). Ex vivo investigation of such events using primary basophils is technically challenging due to the relatively limited number of circulating basophils in peripheral blood. In the current work, the KU812 cell line has been applied as an in vitro model for basophil activation to investigate CARPA-related responses following exposure to test materials obtained from the REFINE consortium. To that end, we developed a standard operating procedure measuring a panel of cell-surface markers indicative of basophilic activation. Two laboratories performed the assays, demonstrating a clear difference in responses between liposomal and polymeric nano(bio)materials, while interlaboratory comparison of the standard operating procedure demonstrated reproducibility in results, between the two facilities. These results suggest the potential to use this protocol as a screening method for such responses however, validation using primary basophils is now warranted.

The interlaboratory comparisons role in the Ukrainian metrology system functioning

It is considered the usage of interlaboratory comparisons for the assuring of national metrology system functioning in the part of national measurement standards recognition (international standards comparisons), development and support the system of the units dissemination (the measurements methods evaluation and reference materials certification by interlaboratory experiment) and the calibration, testing and medical laboratories competence confirmation while their accreditation (proficiency testing). The information on valid European Proficiency Testing System EPTIS is presented. The programs for different types of laboratory studies are considered. The approaches to participants functioning evaluation are presented for the proficiency testing while calibration programs. The recommendations by international documents concerning laboratory activities monitoring reliability are presented.

Automation of evaluation of the interlaboratory comparisons results by means of software

The number of interlaboratory comparisons of measurement results increase, and the number of their participants requires providers to automate data processing procedures. The article is devoted to developing software for the calculation automation and display of interlaboratory comparisons results. The article analyzes the existing software and identifies its main shortcomings. Peculiarities of the application of En, z and ξ criteria are considered. Features of using the Python programming language for calculations and data display are described. Using the example of conducted interlaboratory comparisons, the possibilities of the developed software for automating calculations and data analysis, and displaying results, are demonstrated.

Interlaboratory test results comparisons for active and reactive energy meter

The article is dedicated to the organization and performance of bilateral interlaboratory comparisons of test results for active and reactive energy meter. Interlaboratory comparisons of test results are presented. Evaluation of the laboratory test results bias is provided by means of functioning statistics En criterion. Analysis and conclusions concerning laboratory competence are obtained.

The first total reflection X-ray fluorescence round-robin test of rat tissue samples: Preliminary results

The purpose of the study was performing a round-robin test for the element analysis of mammalian tissue samples using total reflection X-ray fluorescence (TXRF). To achieve this goal, the selected rat organs (kidney, heart, spleen and lung) were subjected to microwave digestion and, in the liquid form, were sent to four European laboratories participating in the inter-comparison investigation. There, using various commercially available TXRF spectrometers, their elemental composition was determined. The carried inter-laboratory comparisons involved macroelements with the concentration above 1000 μg/g (P, S and K) as well as microelements (Ca) and trace elements (Fe, Cu, Zn and Se) occurring, respectively, in the concentrations of 100–1000 μg/g and below 100 μg/g. The validation parameters such as the detection limits of the examined elements as well as the intra-day and inter-day precision were determined. To verify the variation in the results acquired in participating laboratories, for all measured elements, the inter-laboratory precision was evaluated. The data obtained by TXRF were also compared with the ones resulting from ICP-MS (Se) and ICP-OES analysis (P, S, K, Ca, Fe, Cu and Zn).The results obtained as part of the study confirmed the high usefulness of the TXRF method in the elemental analysis of animal tissues. As expected, the best results in terms of the examined validation parameters were obtained for elements with higher atomic numbers (Fe, Cu, Zn and Se). For these analytes, the round-robin test confirmed a good accuracy (around 100% for Fe, Cu, Zn) and precision (intra-day <6%, inter-day <12% and inter-laboratory <12%) what supports the use of TXRF for their determination in the mammalian tissue samples. The obtained results allowed, moreover, to indicate potential issues that require resolving in order to increase the usefulness of the method in case of the analysis of light elements such as P, S or K. It is worth mentioning here, inter alia, the improvement of the sample preparation procedures aiming at the reduction of self-absorption in its dried residue, optimization and/or unification of the sensitivity calibration and the spectra fitting procedures in particular laboratories.

Interlaboratory comparisons of cyanotoxin analysis by ELISA and LC–MS/MS

AbstractTwo commonly used methods for cyanotoxin analysis are enzyme‐linked immunosorbent assay (ELISA) and liquid chromatography/tandem mass spectrometry (LC–MS/MS). Two rounds of interlaboratory comparisons of ELISA and LC–MS/MS analyses were conducted with 12 participating laboratories to evaluate method performances in various matrices, including cyanobacterial bloom and drinking water samples. Fifteen cyanotoxins, including 12 microcystin variants, nodularin, anatoxin‐a, and cylindrospermopsin were evaluated. The impact of sample matrices, preservatives, and quenching reagents was assessed, and no substantial effects were observed. Overall, comparable results were obtained among laboratories performing ELISA and LC–MS/MS analyses, respectively. ELISA results for fortified samples matched more closely with those from LC–MS/MS when microcystin cross‐reactivities were considered, providing data 26% closer to theoretical values on average. This study demonstrates that understanding the effect of cross‐reactivities when comparing ELISA and LC–MS/MS results and considering potential variabilities in commercial standards is important when interpreting data from these two methods.

Interlaboratory comparison study of a chemical profiling method for methylphosphonic dichloride, a nerve agent precursor

An interlaboratory evaluation of a chemical profiling method for a nerve agent precursor was performed by eight chemical analytical laboratories with a wide global distribution. A set of chemical attribution signatures (i.e. impurities) present in methylphosphonic dichloride (DC) were analysed by gas chromatography-mass spectrometry (GC/MS). The GC/MS analyses were performed with the individual instrumentation and analytical methods used by the laboratories in proficiency tests organized by the Organisation for the Prohibition of Chemical Weapons (OPCW). Two batches of DC, produced by two different production routes, were dispatched together with one reference sample. By the use of a targeted MS-library, sixteen chemical attribution signatures were analysed in the DC samples. Retention indices, mass spectra and relative peak areas of the GC/MS data were evaluated. When the within batch-data of the eight laboratories were compared, similarity values of 0.720–0.995 were obtained. In addition, the two different batches had clearly unique chemical attribution profiles, as indicated by a large between batch-distance similarity values of 0.509–0.576. Retention indices showed less than ± 14 retention index unit variation for the 16 chemical attribution signatures. This study showed the potential of getting consistent chemical profiling data over multiple laboratories by the use of retention indices for alignment of GC/MS-data. The results indicate that the GC/MS instrumentation and methods used at most OPCW designated laboratories are valid tools for the acquisition of chemical profiling data of importance for the attribution analysis to study suspected production or use of chemical warfare agents.

External Quality Assurance Schemes (EQUASs) and Inter-laboratory Comparison Investigations (ICIs) for human biomonitoring of polycyclic aromatic hydrocarbon (PAH) biomarkers in urine as part of the quality assurance programme under HBM4EU

Polycyclic aromatic hydrocarbons (PAHs) were included as priority substances for human biomonitoring (HBM) in the European Human Biomonitoring Initiative (HBM4EU), which intended to harmonise and advance HBM across Europe. For this project, a specific Quality Assurance and Quality Control (QA/QC) programme applying Inter-laboratory Comparison Investigations (ICIs) and External Quality Assurance Schemes (EQUASs) was developed to ensure the comparability and accuracy of participating analytical laboratories. This paper presents the results of four ICI/EQUAS rounds for the determination of 13 PAH metabolites in urine, i.e. 1-naphthol, 2-naphthol, 1,2-dihydroxynaphthalene, 2-, 3- and 9-hydroxyfluorene, 1-, 2-, 3-, 4- and 9-hydroxyphenanthrene, 1-hydroxypyrene and 3-hydroxybenzo(a)pyrene. However, 4 PAH metabolites could not be evaluated as the analytical capacity of participating laboratories was too low. Across all rounds and biomarkers, 86% of the participants achieved satisfactory results, although low limits of quantification were required to quantify the urinary metabolites at exposure levels of the general population. Using high-performance liquid or gas chromatography coupled with mass spectrometry (HPLC-MS; GC-MS) and isotope dilution for calibration as well as performing an enzymatic deconjugation step proved to be favourable for the accurate determination of PAHs in urine. Finally, the HBM4EU QA/QC programme identified an international network of laboratories providing comparable results in the analysis of urinary PAH biomarkers, although covering all parameters initially selected was still too challenging.

Detection of adulterated meat products by a next-generation sequencing-based metabarcoding analysis within the framework of the operation OPSON X: a cooperative project of the German National Reference Centre for Authentic Food (NRZ-Authent) and the competent German food control authorities

The German National Reference Centre for Authentic Food (NRZ-Authent) and the competent German food control authorities of the federal states cooperated within the framework of the 10th joint Europol INTERPOL operation OPSON (OPSON X) in the detection of adulterated meat products. A total of 63 meat product samples were collected and analysed by the authorities using standard analytical procedures and subjected to a recently published 16S rDNA metabarcoding analysis. The sequence reads were analysed using 3 bioinformatics data processing strategies. The study aimed to gain additional data on the test samples regarding the authenticity of the declared species and to validate the 16S rDNA metabarcoding method with representative samples. The method was tested not only on 63 test samples, but also on 5 commercial samples from 2 interlaboratory comparison studies and 9 mock mixtures in parallel. The 16S rDNA metabarcoding method was able to detect species that were not target species of the used standard analytical methods, but failed, as shown previously, to detect fallow deer. Otherwise, the qualitative results of the 16S rDNA metabarcoding method were very similar to those of the methods currently in use by the German food control laboratories. Thus, the method has great potential to be used as a screening method for the authentication of mammal and poultry species in meat products.

Data quality control considerations in multivariate environmental monitoring: experience of the French coastal network SOMLIT

IntroductionWhile crucial to ensuring the production of accurate and high-quality data—and to avoid erroneous conclusions—data quality control (QC) in environmental monitoring datasets is still poorly documented.MethodsWith a focus on annual inter-laboratory comparison (ILC) exercises performed in the context of the French coastal monitoring SOMLIT network, we share here a pragmatic approach to QC, which allows the calculation of systematic and random errors, measurement uncertainty, and individual performance. After an overview of the different QC actions applied to fulfill requirements for quality and competence, we report equipment, accommodation, design of the ILC exercises, and statistical methodology specially adapted to small environmental networks (&amp;lt;20 laboratories) and multivariate datasets. Finally, the expanded uncertainty of measurement for 20 environmental variables routinely measured by SOMLIT from discrete sampling—including Essential Ocean Variables—is provided.Results, Discussion, ConclusionThe examination of the temporal variations (2001–2021) in the repeatability, reproducibility, and trueness of the SOMLIT network over time confirms the essential role of ILC exercises as a tool for the continuous improvement of data quality in environmental monitoring datasets.

A ring trial to harmonize Toxoplasma gondii microsatellite typing: comparative analysis of results and recommendations for optimization

A ring trial among five European laboratories was organized to reach consistency in microsatellite (MS) typing of the zoonotic parasite Toxoplasma gondii. Three sample sets were circulated and analyzed by each laboratory following a previously published method that is based on fragment length polymorphism of 15 MS markers. The first sample set compared typing results in general and focused on effects of DNA concentration; the second sample set focused on the polymorphic fingerprinting markers that can differentiate T. gondii strains within the same archetypal lineage; and the third set focused on non-archetypal genotypes. Methodological variations between laboratories, including the software programs used to determine MS fragment length, were collated using a questionnaire. Overall, lineage-level typing results reached a high level of agreement, especially in samples with the highest DNA concentrations. However, laboratory-specific differences were observed for particular markers. Major median differences in fragment length, of up to 6 base pairs, were related to the fluorophore used to label fragment-specific primers. In addition, primer pairs with identical sequences obtained from different suppliers resulted in fragments of differing length. Furthermore, differences in the way the sequencing profiles were assessed and interpreted may have led to deviating results in fragment length determination. Harmonization of MS typing, for example, by using the same fluorophores or by numerical adjustments applied to the fragment-lengths determined, could improve the uniformity of the results across laboratories. This is the first interlaboratory comparison, providing guidelines (added as a supplement) for the optimization of this technique.

Ex vivo instability of lipids in whole blood: preanalytical recommendations for clinical lipidomics studies

Reliability, robustness, and interlaboratory comparability of quantitative measurements is critical for clinical lipidomics studies. Lipids’ different ex vivo stability in blood bears the risk of misinterpretation of data. Clear recommendations for the process of blood sample collection are required. We studied by UHPLC-high resolution mass spectrometry, as part of the “Preanalytics interest group” of the International Lipidomics Society, the stability of 417 lipid species in EDTA whole blood after exposure to either 4°C, 21°C, or 30°C at six different time points (0.5 h–24 h) to cover common daily routine conditions in clinical settings. In total, >800 samples were analyzed. 325 and 288 robust lipid species resisted 24 h exposure of EDTA whole blood to 21°C or 30°C, respectively. Most significant instabilities were detected for FA, LPE, and LPC. Based on our data, we recommend cooling whole blood at once and permanent. Plasma should be separated within 4 h, unless the focus is solely on robust lipids. Lists are provided to check the ex vivo (in)stability of distinct lipids and potential biomarkers of interest in whole blood. To conclude, our results contribute to the international efforts towards reliable and comparable clinical lipidomics data paving the way to the proper diagnostic application of distinct lipid patterns or lipid profiles in the future.

Myo-Inositol, Scyllo-Inositol, and Other Minor Carbohydrates as Authenticity Markers for the Control of Italian Bulk, Concentrate, and Rectified Grape Must.

Myo-inositol polyalcohol is a characteristic component of natural and concentrated grape musts (CMs), and Regulation (EU) no. 1308/2013 prescribes its presence as a marker of the authenticity of rectified concentrated must (RCM). Other polyalcohols besides myo-inositol, such as scyllo-inositol or minor sugars, could be considered authenticity markers, but an extensive search in the literature yielded no exhaustively investigated study of their concentration variability in genuine products. The aim of this study was to create an extensive national data bank of minor carbohydrates profiles and investigate the impact of the geographical origin and the different vintages on the concentration of these compounds; to this end, 450 authentic Italian grape musts of different varieties were sampled and analyzed during the harvest season in 2019, 2020, and 2021. The grape musts from the Italian wine-growing areas CII and CIIIb had myo- and scyllo-inositol contents always higher than 756 and 39 mg/kg of sugar, respectively. Conversely, also considering other mono- and disaccharides, sucrose, sorbitol, lactose, maltose, and isomaltose showed contents always lower than 534, 1207, 390, 2222, and 1639 mg/kg of sugar, respectively. The general applicability to the CM and RCM of the proposed authenticity thresholds, established in the must, was demonstrated by studying the influence of must concentration on the myo- and scyllo-inositol content. Inter-laboratory comparison experiments were also conducted to harmonize and characterize laboratory methods and validate the analytical dataset. Based on the obtained results, the text of the EU legislation (Reg. (EU) 1308/2013), which defines the characteristics of the must and the CRM product, should be revised.