Confirmatory Method Research Articles

The challenge of colorectal cancer (CRC) screening in low and middle-income countries (LMIC): The Brazilian ten year long experience.

129 Background: The increasing incidence of CRC seen in LMIC and the high mortality rates compared to high-income countries points towards the difficult implementation of an effective screening strategy in LMIC. Brazil is one of the few Latin American countries and LMIC that have a population-based CRC screening program, targeting ages 50 to 74 and performing a fecal occult blood test (FT) every 2 years and colonoscopy/sigmoidoscopy as a confirmatory test. Methods: We performed a cross-sectional study, which evaluated the prevalence of every stage of CRC/carcinoma in situ (CIS) at the time of its diagnosis from 2013 to 2022. Similar data regarding the Brazilian breast and cervical cancer screening programs was obtained. Each stage was evaluated by age targeted by their respective screening program using the national database (DATASUS). Results: In the analyzed decade there was a total of 87507 CRC/CIS cases in the overall studied population, which only 11% was of Stage I or less and 70% was of stage III or IV at the time of diagnosis. In the subdivided age groups, the prevalence of each stage remained somewhat constant through all groups (In situ/Stage 0 = 8-9%; Stage I = 3%; Stage II = 20-21%; Stage III = 35-36%; Stage IV = 32-34%) and the majority of cases occurred from age 60 to 64 (20549 cases). In 2022, a total of 2128168 RT was performed - corresponding to only 4,6% of the targeted population - and 408489 colonoscopies. When compared to other cancer screening programs, CRC presents indifferent to age group, sustaining constant rate of regional/distant stages, with at least 32% of patients presenting a <17% 5-year survival rate (5-YSR), whereas cervical cancer presents approximately 80% of cases at >91% 5-YSR at the first age group, and breast cancer with 89% of cases at >86% 5-YSR at the first age group. Conclusions: Data suggests the inefficacy of CRC screening implementation, and literature sustains to be due to its poor acceptance by the population and the unavailability of confirmatory imaging tests due to long-waiting times and high cost, reducing its ability to detect early stage lesions, in contrast with the overall acceptance and high effectivity of cervical/breast cancer screening. The discrepancy between the guidelines and their implementation as a screening program in LMIC are in consonant. This study aims to be a launching point for new research and interventional studies evaluating the impact of better locally adjusted actions in the prevention of CRC such as screening invitations, in-community health education, and the developing of better cost-benefit confirmatory methods might hinder the advance of CRC in LMIC.[Table: see text]

T-2 and HT-2 Toxins: Toxicity, Occurrence and Analysis: A Review.

One of the major classes of mycotoxins posing serious hazards to humans and animals and potentially causing severe economic impact to the cereal industry are the trichothecenes, produced by many fungal genera. As such, indicative limits for the sum of T-2 and HT-2 were introduced in the European Union in 2013 and discussions are ongoing as to the establishment of maximum levels. This review provides a concise assessment of the existing understanding concerning the toxicological effects of T-2 and HT-2 in humans and animals, their biosynthetic pathways, occurrence, impact of climate change on their production and an evaluation of the analytical methods applied to their detection. This study highlights that the ecology of F. sporotrichioides and F. langsethiae as well as the influence of interacting environmental factors on their growth and activation of biosynthetic genes are still not fully understood. Predictive models of Fusarium growth and subsequent mycotoxin production would be beneficial in predicting the risk of contamination and thus aid early mitigation. With the likelihood of regulatory maximum limits being introduced, increased surveillance using rapid, on-site tests in addition to confirmatory methods will be required. allowing the industry to be proactive rather than reactive.

Chemical and Toxicological Diagnosis of Acute Poisoning with Doxylamine, Zaleplon, and Phenazepam

BACKGROUND The market of hypnotic and sedative drugs is being updated due to the high toxicity of barbiturates and the limitations of their use. Currently, safer drugs such as Z-drugs, Doxylamine, and some benzodiazepine derivatives are often prescribed for the treatment of anxiety and insomnia, but they can cause acute poisoning if overdosed or in case of nonmedical use.AIM To establish an affordable express thin-layer chromatography (TLC) technique for preliminary screening detection of Doxylamine, Phenazepam and Zaleplon in order to diagnose acute poisoning.MATERIAL AND METHODS Thin-layer chromatography (TLC) and gas chromatography with mass selective detection (GC-MS) methods were used. Urine samples from patients with symptoms of acute Doxylamine, Zaleplon, Phenazepam poisoning, and model urine samples were prepared by liquid-liquid extraction at pH 9.0 with chloroform for TLC analysis, with ethyl acetate-diethyl ether mixture (1:1) for GC-MS.RESULTS We developed the TLC method of Doxylamine, Zaleplon and Phenazepam detection which helps reveal their presence in the patient’s urine, as well as distinguish one from another in case of similar toxic symptoms. The GC-MS method was used for confirmatory analysis. Compared to confirmatory methods, the developed technique of TLC screening is expressive, does not require expensive high-tech equipment, while allowing to differentiate Doxylamine, Zaleplon and Phenazepam from each other and from other toxicologically significant psychoactive substances detected in general screening.

Evaluation of The Performance of Incubation Business Services in Morocco

The purpose of this article is to make an empirical assessment of the services offered by the incubation structures in Morocco. To do this, we will focus on the identification of the incubation actors and the services offered by these structures. Within this framework, two studies are envisaged: the first one aims at exploring entrepreneurial incubation as it is conceived and seen by its practitioners, and a confirmatory method that will focus on the perception of this practice on the part of "incubated or clients of incubators". The results are complementary and indicate a gap between the image of incubation and its reality. This has helped us to formulate recommendations to bring our judgment on this practice.

Psychometric analysis of a tool to assess metamotivational strategies in undergraduate anesthesia students

Background & Objective: Undergraduate anesthesia students benefit from motivational self-regulation strategies when facing specific academic challenges to maintain and improve their academic motivation. Due to the absence of valid and reliable instruments to assess these strategies, the present study aimed at the psychometrical analysis of MSMQ (Metamotivational Strategies in Medical Students) which is used to measure meta-motivational strategies in undergraduate anesthesia students. Materials & Methods: In this psychometric study, 358 undergraduate anesthesia students were selected by convenience sampling method. The research community included undergraduate anesthesia students in Iran, and the MSMQ questionnaire was provided for them to complete. Its validity was determined by face and content validity methods, and its reliability was determined by the internal consistency method. The confirmatory factor analysis method was used to check construct validity. Results: The CVI and CVR values were obtained at ≥ 0.80 for all items. In the confirmatory factor analysis, the factor loading of all items was reported to be ≤ 0.40, and the mentioned instrument enjoyed the goodness of fit indices (RMSEA = 0.064, NNFI = 0.96, CFI = 0.97, SRMR = 0.06, GFI = 0.86). Acceptable reliability coefficients were obtained for the subscales, and Cronbach's alpha coefficient for the whole questionnaire was 0.922. Conclusion: As evidenced by the results of this study, the MSMQ questionnaire is a valid and reliable instrument; therefore, it can be used for Iranian undergraduate anesthesia students.

Assessing pharmaceuticals in the green seaweed Ulva lactuca through a multi-residue UHPLC-ToF-MS strategy

Seaweeds have become an important asset in several sectors, including the food and feed industries, cosmetics, and pharmaceuticals, among others. Whether harvested or reared, interest in algae has been growing worldwide due to the resources they offer, including proteins, vitamins, minerals, carbohydrates, essential fatty acids, and dietary fiber, as well as sources of biologically active compounds. However, given their morphology and physiology, as well as their harvest and cultivation environments, algae are prone to the presence of hazards, including pharmaceuticals taken up from the water. Thus, to ensure human and animal safety as well as environmental health, monitoring is essential. Therefore, this work describes the development and validation of a sensitive screening and confirmatory analytical method based on ultra-high-performance liquid chromatography coupled with time-of-flight mass spectrometry (UHPLC-ToF-MS). This multi-residue method enables the determination of 62 pharmaceuticals distributed between 8 therapeutic classes and was fully validated according to Commission Implementing Regulation (EU) 2021/808.

Parkinson’s disease. Focus on early stages

The review presents current data on the peculiarities of the neurodegenerative process in the early stages of Parkinson’s disease (PD) and considers the hypothesis of the presence of body-first and brain-first subtypes of the disease onset. The earliest manifestations of the disease include symptoms such as parasomnia, constipation, hyposmia, anxiety-depressive disorder, daytime sleepiness, color perception changes, cognitive dysfunction, and mild motor manifestations. The diagnosis of PD can be made when characteristic motor manifestations occur: hypokinesia, rest tremor, muscle rigidity. Substantia nigra ultrasound, magnetic resonance imaging in SWI mode, and functional neuroimaging methods are used as confirmatory methods. The approach to the treatment of early stages of PD is age dependent. Patients over 70 years of age are recommended to start therapy with levodopa; younger patients – with dopamine receptor agonists (ADR), amantadines, MAO-B inhibitors. The mechanisms of ADR action, the possibility of their influence on the motor and non-motor symptoms of PD are analyzed. A special place is given to piribedil, which, due to the dual mechanism of action, has a positive effect on cognitive functions, depression, apathy, as well as a minimal effect on daytime sleepiness.

Characteristics of Extended-Spectrum β-Lactamase-Producing Escherichia coli Derived from Food and Humans in Northern Xinjiang, China.

This study aimed to investigate the drug resistance, molecular characteristics, and genetic relationship of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolated from food and human stool samples in northern Xinjiang. From 2015 to 2016, a total of 431 samples (meats and vegetables) were collected from retail markets and supermarkets located in the regions of Urumqi, Shihezi, and Kuitun in Xinjiang, China, and 20 human stool samples from the Shihezi Hospital. The PCR method was used to detect E. coli, and the presence of ESBL-producing E. coli was confirmed using the K-B disk diffusion confirmatory method. The susceptibility to ESBL-producing E. coli was tested by the microdilution broth method, and the minimum inhibitory concentration was determined. PCR was used to detect the resistance and virulence genes of ESBL-producing E. coli, and phylogenetics, plasmid replicon typing, screening of three integrons, and multilocus sequence typing (MLST) were performed. The results showed that 127 E. coli strains (15 human stool and 112 food samples) were isolated. Out of the 127 E. coli strains, 38 strains (6 human stool and 32 food 34 samples) of ESBL-producing E. coli were identified through screening. These 38 strains showed resistance to cefotaxime (94.74%) and cefepime (94.74%), and were sensitive to meropenem (0.00%). The most detected resistance genes were blaTEM (47.37%), and the most detected virulence genes were fimH (97.73%), ompA (97.73%), hlyE (97.73%), and crl (97.37%). The isolates belonged to phylogroups B1 (42.11%), C (23.68%), and A (21.05%). Among the plasmid replicon subtypes, IncFIB was the main type (42.11%). The integrons detected were of the first type (47.37%) and the third type (26.32%). The 38 E. coli strains had 19 different sequence-type (ST) strains. These 38 strains of ESBL-producing E. coli were analyzed using MLST and STs are varied.

Simplified Liquid Chromatography–Mass Spectrometry Methods for Gestagen Analysis in Animal Fat and Liver

Gestagens, a class of veterinary drugs also called progestogens, are synthetic hormones used to increase feed efficiency and rate of gain in heifers. The Canadian Food Inspection Agency analyzes progestogens melengestrol acetate (MGA), megestrol acetate, and chlormadinone acetate using liquid chromatography–mass spectrometry (LC–MS). Our conventional gestagen method for kidney fat has many time-consuming steps, including solid-phase extraction. A sample preparation procedure having fewer clean-up steps was developed for routine diagnostic analysis of kidney fat and provided similar results faster, and at lower cost. A confirmatory liver method for gestagens, developed using salt-assisted extraction, employed minimal clean-up steps that resulted in high chemical background at the desired lower limit of quantification (LLOQ). Differential ion mobility spectrometry, specifically high-field asymmetric waveform ion mobility spectrometry (FAIMS), was used to filter chemical background in the gas phase. The effect of the ionization probe position on FAIMS parameters, including sensitivity, is described. With LC-FAIMS-MS, chemical background for each gestagen was virtually eliminated, resulting in a quantitative liver method having the desired 0.6 ng/g LLOQ and estimated limits of detection (LODs) up to 140 times lower than LC-MS. Incurred MGA samples, analyzed using kidney fat and liver methods from the same animal, show levels within the quantitative ranges of both methods.

Sex differences in the psychometric properties of the Center for Epidemiological Studies-Depression (CES-D) Scale in older Filipinos.

The literature has yet to fully account for sex differences in the symptomatology and measurement of depressive symptoms, especially in developing settings like the Philippines. Thus, we established the factor structure and assessed the reliability of the 11-item version of the Center for Epidemiological Studies-Depression (CES-D) Scale for assessing depressive symptoms in older Filipino men and women. Using cross-sectional data from 5,209 community-dwelling Filipinos aged 60 and above from a nationally representative survey, Confirmatory Factor Analysis (CFA) and methods in Item Response Theory (IRT) were applied to provide complementary insights into the properties of the scale and its individual items. CFA supported the multidimensionality of the scale. The scale is also sex-invariant, but the relationship between the subfactors and the higher-order factor may differ between men and women. In addition, findings from IRT analysis confirmed the overall utility of the CES-D scale, but positively stated items were found to be internally inconsistent with the rest of the scale. The scale is desirably informative for assessing more severe symptoms, although sex differences were found in the precision of individual items. In general, the 11-item CES-D Scale is an adequate multidimensional tool for assessing moderate to severe depressive symptoms in the older population, especially in older men.

Determination of quinolones, macrolides, sulfonamides and tetracyclines in honey using QuEChERS sample preparation and UHPLC-MS/MS analysis

Monitoring beehive health and timely prevention of possible infections by bacteria, mould, viruses or parasites is exceptionally important in beekeeping. Antibiotics originating from the environment can be found in honey and due to improper beekeeping practices. Enabling the detection of antibiotic residues in honey and suppressing the development of antibiotic resistance require the development of a sensitive multi-class method for the determination of antibiotics. For the purpose of analysing honey, a screening and confirmatory method for the determination of 36 antibiotics was developed. The QuEChERS procedure and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was selected to achieve high sensitivity and selectivity. Validation according to the new Commission Implementing Regulation (EU) 2021/808 included the following performance characteristics: selectivity, trueness, precision, decision limit (CCα), detection capability (CCβ) and relative matrix effect. The method was validated in the measurement range from 1 to 10 μg/kg, where maximum trueness and acceptable coefficients of variation were achieved. The detection capability (CCβ) equals the lowest concentration level for all analytes, ranging from 0.1 μg/kg to 2.5 μg/kg, as the false compliant rate less than 5% was confirmed at this level of interest. For unauthorised pharmacologically active substances, the CCα was determined and ranged from 0.16 μg/kg for sulfaquinoxaline to 3.67 μg/kg for difloxacin. The high matrix influence of floral and chestnut honey indicated the need for quantitative analysis using the matrix calibration curve.

Confirmatory evaluation of eFUME for control of brown marmorated stink bug, Halyomorpha halys (Hemiptera: Pentatomidae).

The governments of Australia and New Zealand require a phytosanitary treatment to control adult brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), as overwintering aggregations have been intercepted in the importation pathway of various nonhorticultural consignments, including vehicles. The commercialized fumigant, eFUME, a 16.7% by mass dilution of ethyl formate in carbon dioxide, resulted in complete control of ca. 1,000 field-collected, naturally diapausing adult H. halys in each of 3 independent trials at 10 ± 0.5 °C (x¯±2s) when ethyl formate levels in enclosure headspace were maintained steady-state at ca. 14.5 mg/liter for 4 h to yield Ct exposures ranging from 57.9 to 63.1 mg/liter h. Consistent with previous findings where greenhouse reared H. halys were controlled using laboratory formulations of this ethyl formate-carbon dioxide mixture, these confirmatory methods and results further inform technical and operational features of commercial practice.

DART-HRMS allows the detection of toxic alkaloids in animal autopsy specimens and guides the selection of confirmatory methods in accidental plant poisoning

BackgroundIn cases of suspected animal poisonings or intoxications, there is the need for high-throughput, rapid and accurate analytical tools capable of giving rapid answers and, thus, speeding up the early stages of investigations. Conventional analyses are very precise, but do not meet the need for rapid answers capable of orienting the decisions and the choice of appropriate countermeasures. In this context, the use of ambient mass spectrometry (AMS) screening methods in toxicology laboratories could satisfy the requests of forensic toxicology veterinarians in a timely manner. ResultsAs a proof of principle, direct analysis in real time high resolution mass spectrometry (DART-HRMS) was applied to a veterinary forensic case in which 12 of a group of 27 sheep and goats died with an acute neurological onset. Because of evidence in the rumen contents, the veterinarians hypothesized an accidental intoxication after ingestion of vegetable materials.The DART-HRMS results showed abundant signals of the alkaloids calycanthine, folicanthidine and calycanthidine, both in the rumen content and at the liver level. The DART-HRMS phytochemical fingerprinting of detached Chimonanthus praecox seeds was also compared with those acquired from the autopsy specimens. Liver, rumen content and seed extracts were then subjected to LC-HRMS/MS analysis to gather additional insights and confirm the putative assignment of calycanthine anticipated by DART-HRMS.HPLC-HRMS/MS confirmed the presence of calycanthine in both rumen contents and liver specimens and allowed its quantification, ranging from 21.3 to 46.9 mg kg−1 in the latter. This is the first report detailing the quantification of calycanthine in liver after a deadly intoxication event. Significance and noveltyOur study illustrates the potential of DART-HRMS to offer a rapid and complementary alternative to guide the selection of confirmatory chromatography-MSn strategies in the analysis of autopsy specimens from animals with suspected alkaloid intoxication. This method offers the consequent saving of time and resources over those needed for other methods.

Modern approaches to cervical cancer screening (a review)

The aim – systematization of data from modern scientific sources and own knowledge and experience regarding the problems, advantages and prospects of the development of various methods of diagnosing pathological conditions of the cervix. The relevance of the problem of cervical cancer is beyond doubt, which is confirmed by the annual data of the official statistics and recommendations of WHO, European experts and relevant national recommendations. The analysis and generalization of information from the specialized literature regarding the assessment of problems and prospects for the development of leading methods of cervical screening in order to reduce the number of cervical cancer cases in the world is a priority task of the expert society. Currently, the most recommended approach to cervical screening is to combine HPV testing with cytology but separate testing for HPV is the most promising. Colposcopy remains the only confirmatory method of diagnosing cervical pathology, subject to mandatory cervical biopsy and histopathological examination of the biopsy. Standardization of classifications and terminology in the assessment of cytological, histopathological and colposcopic conclusions will allow to reach an understanding in the issues of optimal management tactics for patients with cervical pathology. Conclusions. Diagnosis of high-risk carcinogenic types of human papillomavirus is the most promising method of cervical screening in the near future. At the same time, at the moment, we consider it appropriate to use the cytology together with HPV testing, at least as a sorting method. Standardization of cytological/histopathological terminology should be done in accordance with the Bethesda 2014 system update.

Deep learning-based semantic segmentation of non-melanocytic skin tumors in whole-slide histopathological images.

Basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are the two most common skin cancer and impose a huge medical burden on society. Histopathological examination based on whole-slide images (WSIs) remains to be the confirmatory diagnostic method for skin tumors. Accurate segmentation of tumor tissue in WSIs by deep-learning (DL) models can reduce the workload of pathologists and help surgeons ensure the complete removal of tumors. To accurately segment the tumor areas in WSIs of BCC, SCC and squamous cell papilloma (SCP, homologous to SCC) with robust models. We established a data set (ZJU-NMSC) containing 151 WSIs of BCC, SCC and SCP in total. Seven models were utilized to segment WSIs, including the state-of-the-art model, models proposed by us and other models. Dice score, intersection over union, accuracy, sensitivity and specificity were used to evaluate and compare the performance of different models. Heatmaps and tumor tissue masks were generated to reflect the results of the segmentation. The processing times of models are also recorded and compared. While the dice score of most models is higher than 0.85, deeplab v3+ has the best performance and the corresponding tumor tissue mask is more consistent with the ground truth tumor areas even with complex and small lobular lesions. This study broadens the use of DL-based segmentation models in WSIs of skin tumors in terms of tumor types and computational approaches. Segmenting tumor areas can simplify the process of histopathological inspection and benefit the diagnosis and following management of the diseases in practice.

Molecular immunological techniques to support the differential microbiological diagnosis of acute HIV infection

This research sought to identify HIV confirmatory molecular immunological methods for the early diagnosis of acute infections, complementing the differential microbiological diagnosis. A descriptive study of a clinical case was carried out using molecular immunological analyses. The results showed that molecular immunological methods range from third generation techniques, capable of diagnosing the infection in the acute phase, to fourth generation techniques that detect the infection from the third week of infection. Confirmatory tests, such as Western Blot and LIA, presented high sensitivity and specificity. Revised molecular techniques, such as p24 detection or Polymerase Chain Reaction (PCR), reduced diagnosis time. In conclusion, molecular immunological methods allow early detection of HIV with high precision, providing diagnostic and confirmation results from the third week of infection.

Financial Planning for Retirement: The Moderating Role of Government Policy

The objective of this study is to examine the role of financial self-efficacy (FSE), retirement goal clarity (RGC), and government policy (GP) on financial planning for the retirement (FPR) behavior of public university employees based on the CWO model for retirement. Also, to examine the moderating role of GP on the relationship between FSE, RGC, and FPR. Primary data was gathered during the COVID-19 pandemic from mid-July 2020 until the end of January 2021 using a non-probability convenience sampling approach involving 525 respondents. The data were analyzed using the Structural Equation Modelling (SEM) technique. The Confirmatory Tetrad Analysis method (CTA-PLS) is used to determine the formative/reflective type of study variables. The results directly show the significant impact of the FSE, RGC, and GP on FPR. Indirectly, for those with a clear retirement goal, the perceived GP moderates its relationship with FPR. Previous research has examined FSE and RGC in FPR without considering the effect of GP.

An assessment of strategies used on resources mobilization: A case of the Université Chrétienne Bilingue Du Congo (UCBC)

Many people in institutions or organizations have excellent visions and initiatives but most ofthem don’t see their visions come true because of either lack of resources or mismanagement ofthe mobilized resources. This study focuses on an assessment of strategies used on resourcesmobilization in the Université Chrétienne Bilingue du Congo (UCBC). In particular, the studysought to find out whether community participation, local fundraising, management practicesand marketing strategies affect resource mobilization at UCBC. The study adopted a quantitativeapproach targeting a population of 509 subjects. Stratified and purposive sampling techniqueswere used to get a sample size of 95 subjects. Data was collected using structured questionnairewith Likert scale measurement. The collected data was analyzed using confirmatory factoranalysis method. The results found out that management practices (p-value 0.0018), marketingstrategies (p-value 0.0430), local fundraising (p-value 0.0849) and community participation (pvalue 0.0019) were positive and significantly related to resource mobilization. The study recommends that UCBC considers the significant variables as they affect resource mobilization performance.

IMPLEMENTATION OF THE SYSTEM OF HONEY SAFETY CONTROL ON THE EXAMPLE OF ANALYZING OF RESIDUES OF TETRACYCLINE GROUP ANTIBIOTICS

The National Reference Laboratory of Drug Residues Control has implemented a system of "classical" honey safety control based on the parameters of antimicrobial drug residues: samples testing by the screening enzyme-linked immunosorbent assay (ELISA) method and, in the case of positive or possibly positive samples occurring, – retesting by confirmatory UPLC-MS/MS multi-method developed in the laboratory. The article presents the results of ELISA screening method validation for the determination of residual amounts of tetracycline group antibiotics (tetracycline, oxytetracycline, chlortetracycline, doxycycline) in honey samples using Tetracyclin® RIDASCREEN test kits by R-Biopharm (Germany) and Oxytetracycline ELISA by EuroProxima (the Netherlands) and its verification by means of the confirmatory method.
 In accordance with the Commission Implementing Regulation (EU) 2021/808, the validation of test kits proposed by the manufacturers was carried out taking into account the minimum required limit of detection. The fitness-for-purpose of the methods was confirmed based on the establishment of main validation parameters for screening methods (technical threshold and cut-off factor) using control (blank) and fortified honey samples with standard solutions of tetracycline at the level of ½ of the minimum required limit of detection, according to the "added-received" criterion. The proposed methods are the simple in carrying out, time- and cost-effective, which are their main advantages.
 The reliability of the results obtained by the proposed methods was verified using the developed confirmatory UPLC-MS/MS multi-method. The results of comparative studies of control (blank) honey samples and fortified with a standard solution of tetracycline antibiotics at the level of 2.5 and 5.0 μg/kg, as well as the results of the interlaboratory testing with the accredited laboratory (Germany) are presented.

Development and Validation of a Confirmatory Method for the Determination of 12 Coccidiostat Residues in Eggs and Muscle by Means of Liquid Chromatography Coupled to Hybrid High Resolution Mass Spectrometry

A confirmatory, highly selective multi-residue method based on liquid chromatography coupled to hybrid high resolution mass spectrometry (LC-Q-Orbitrap) was developed and validated for the determination of 12 regulated coccidiostats in eggs and muscle. Particularly, ionophore antibiotics (lasalocid, maduramicin, monensin, narasin, salinomycin and semduramicin) and synthetic coccidiostats (diclazuril, halofuginone, nicarbazin as 4,4′-dinitrocarbanilide fraction, robenidine and toltrazuril as toltrazuril-sulphone) were included in the method. The sample preparation consisted in the extraction of the analytes from the matrix with acetonitrile, followed by a clean-up step with Oasis® PRiME HLB SPE and a defatting procedure with n-hexane. Validation was successfully performed according to Commission Implementing Regulation (EU) 2021/808, starting from 1 µg kg−1. The procedure was verified through the analysis of a certified reference material (CRM) and the occurrence of the residues was assessed in the context of the Italian National Residue Control Plan (NRCP).