Certified reference materials for the determination of mycotoxins

A continuing fundamental trend in the area of quality assurance in analytical laboratories are measures that enable trace-ability of analytical results to internationally renowned units, like e.g. the mole. Especially certified reference materials (CRMs) are important tools for achieving this goal. What are CRMs? Why do we need them in the field of mycotoxin analysis? Which kind of CRMs are currently available for the determination of mycotoxins and how should they be used? These are questions, which are increasingly being asked. This article attempts to answer these questions.     

Mycotoxicoses of domestic animals and their diagnosis

Mycotoxins have been named "agents in search of a disease," and the considerable progress in analytical methodology over the last 10 years has not changed this very much. The following are factors that contribute to the difficulty of making a diagnosis: (1) nonspecificity of lesions; (2) masking of mycotoxic effects by secondary effects, e.g., through immunosuppression; (3) late appearance of a lesion, e.g., bone marrow damage or neoplasia; (4) interaction of several mycotoxins or presence of other toxicants or deficiency states; (5) species variation in the response to the mycotoxin(s); (6) difficulty of linking a late appearing effect with a demonstrable cause; (7) low doses of mycotoxins may cause stimulating effects; and (8) not being aware of the potential of a mycotoxin as a causative factor in disease. The mycotoxins of major importance in Canada are trichothecenes, ochratoxin, zearalenone, and ergot. It is concluded that the significance of mycotoxins for animals in Canada is likely generally underestimated.     

Determination of mycotoxins in foods: current state of analytical methods and limitations

Mycotoxins are natural contaminants produced by a range of fungal species. Their common occurrence in food and feed poses a threat to the health of humans and animals. This threat is caused either by the direct contamination of agricultural commodities or by a “carry-over” of mycotoxins and their metabolites into animal tissues, milk, and eggs after feeding of contaminated hay or corn. As a consequence of their diverse chemical structures and varying physical properties, mycotoxins exhibit a wide range of biological effects. Individual mycotoxins can be genotoxic, mutagenic, carcinogenic, teratogenic, and oestrogenic. To protect consumer health and to reduce economic losses, surveillance and control of mycotoxins in food and feed has become a major objective for producers, regulatory authorities and researchers worldwide. However, the variety of chemical structures makes it impossible to use one single technique for mycotoxin analysis. Hence, a vast number of analytical methods has been developed and validated. The heterogeneity of food matrices combined with the demand for a fast, simultaneous and accurate determination of multiple mycotoxins creates enormous challenges for routine analysis. The most crucial issues will be discussed in this review. These are (1) the collection of representative samples, (2) the performance of classical and emerging analytical methods based on chromatographic or immunochemical techniques, (3) the validation of official methods for enforcement, and (4) the limitations and future prospects of the current methods.     

Factors Affecting Accuracy of Data Abstracted from Medical Records

ObjectiveMedical record abstraction (MRA) is often cited as a significant source of error in research data, yet MRA methodology has rarely been the subject of investigation. Lack of a common framework has hindered application of the extant literature in practice, and, until now, there were no evidence-based guidelines for ensuring data quality in MRA. We aimed to identify the factors affecting the accuracy of data abstracted from medical records and to generate a framework for data quality assurance and control in MRA.MethodsCandidate factors were identified from published reports of MRA. Content validity of the top candidate factors was assessed via a four-round two-group Delphi process with expert abstractors with experience in clinical research, registries, and quality improvement. The resulting coded factors were categorized into a control theory-based framework of MRA. Coverage of the framework was evaluated using the recent published literature.ResultsAnalysis of the identified articles yielded 292 unique factors that affect the accuracy of abstracted data. Delphi processes overall refuted three of the top factors identified from the literature based on importance and five based on reliability (six total factors refuted). Four new factors were identified by the Delphi. The generated framework demonstrated comprehensive coverage. Significant underreporting of MRA methodology in recent studies was discovered.ConclusionThe framework generated from this research provides a guide for planning data quality assurance and control for studies using MRA. The large number and variability of factors indicate that while prospective quality assurance likely increases the accuracy of abstracted data, monitoring the accuracy during the abstraction process is also required. Recent studies reporting research results based on MRA rarely reported data quality assurance or control measures, and even less frequently reported data quality metrics with research results. Given the demonstrated variability, these methods and measures should be reported with research results.     

Co-expression of 15 contiguous genes delineates a fumonisin biosynthetic gene cluster in Gibberella moniliformis

Fumonisins are mycotoxins produced by the maize pathogen Gibberella moniliformis and are associated with cancer in rodents. In this study, we determined the nucleotide sequence of a 75-kb region of G. moniliformis DNA and identified 18 heretofore undescribed genes flanking a cluster of five previously identified fumonisin biosynthetic (FUM) genes. Ten of the newly identified genes downstream of the cluster were coregulated with FUM genes and exhibited patterns of expression that were correlated with fumonisin production. BLASTX analyses indicated that the predicted functions of proteins encoded by the 10 genes were consistent with activities expected for fumonisin biosynthesis or self-protection. These data indicate that the 10 newly identified genes and the previously identified FUM genes constitute a fumonisin biosynthetic gene cluster. Disruption of two of the new genes, encoding longevity assurance factors, had no apparent effect on fumonisin production, but disruption of a third, encoding an ABC transporter, had a subtle effect on ratios of fumonisins produced.     

Case study and application of process analytical technology (PAT) towards bioprocessing: II. Use of ultra-performance liquid chromatography (UPLC) for making real-time pooling decisions for process chromatography

Process analytical technology (PAT) has been gaining a lot of momentum in the biopharmaceutical community due to the potential for continuous real-time quality assurance resulting in improved operational control and compliance. Two of the key goals that have been outlined for PAT are "variability is managed by the process" and "product quality attributes can be accurately and reliably predicted over the design space established for materials used, process parameters, manufacturing, environmental, and other conditions". Recently, we have been examining the feasibility of applying different analytical tools for designing PAT applications for bioprocessing. We have previously shown that a commercially available online high performance liquid chromatography (HPLC) system can be used for analysis that can facilitate real-time decisions for column pooling based on product quality attributes (Rathore et al., 2008). In this article we test the feasibility of using a commercially available ultra- performance liquid chromatography (UPLC) system for real-time pooling of process chromatography columns. It is demonstrated that the UPLC system offers a feasible approach and meets the requirements of a PAT application. While the application presented here is of a reversed phase assay, the approach and the hardware can be easily applied to other modes of liquid chromatography.     

Quantification of glycated IgG in CHO supernatants: A practical approach

Post-translational, nonenzymatic glycation of monoclonal antibodies (mAbs) in the presence of reducing sugars (in bioprocesses) is a widely known phenomenon, which affects protein heterogeneity and potentially has an impact on quality, safety, and efficacy of the end product. Quantification of individual glycation levels is compulsory for each mAb therapeutically applied in humans. We therefore propose an analytical method for monitoring glycation levels of mAb products during the bioprocess. This is a useful tool for process-design considerations, especially concerning glucose-feed strategies and temperature as major driving factors of protein glycation. In this study, boronate affinity chromatography (BAC) was optimized for determination of the glycation level of mAbs in supernatants. In fact, the complex matrix found in supernatants is an underlying obstacle to use BAC, but with a simple clean-up step, we found that the elution profile could be significantly improved so that qualitative and quantitative determination could be reached. Complementary analytical methods confirmed the performance quality, including the correctness and specificity of the results. For quantitative determination of mAb glycation in supernatants, we established a calibration procedure for the retained mAb peak, identified as glycated antibody monomers. For this approach, an available fully characterized mAb standard, Humira®, was successfully applied, and continuous monitoring of mAbs across three repetitive fed-batch processes was finally performed. With this practical, novel approach, an insight was obtained into glycation levels during bioprocessing, in conjunction with glucose levels and product titer over time, facilitating efficient process development and batch-consistency monitoring.     

State-of-the-Art Procedures and Quality Management in Diagnostic Medical Mycology

Detection and correct identification of fungi causing invasive disease is a major challenge for diagnostic laboratories. However, early and reliable diagnosis is essential to keep mortality to a minimum, whereas delayed or missed diagnosis is typically associated with fatal outcome in the case of invasive fungal infections. Therefore, rapid and reliable results are of importance for successful management. In order to guarantee correct reports, accurate processing of all samples and appropriate methods for detecting the fungal agent as well as quality assurance is mandatory. This article gives an overview of current methods and elucidates quality management measures in medical mycology laboratory.     

Mycotoxins: food contamination, mechanism, carcinogenic potential and preventive measures

Mycotoxins constitute a large number of naturally occurring fungal secondary metabolites with very diversified toxic effects in humans and animals. Among many mycotoxins discovered, aflatoxins, ochratoxin A, sterigmatocystin and several others are identified as carcinogens; several others were found to be mutagenic. Nevertheless, aflatoxin B1 has been found to be one of the most potent carcinogens and contamination of aflatoxins in the food supply is still a major concern. Whereas extensive studies have been made on aflatoxins, little is known about the mode of action of other carcinogenic and mutagenic mycotoxins. Recent progress on research for the carcinogenic and mutagenic mycotoxins is presented in this review with emphasis on their contamination in foods, their carcinogenic potential to humans, and the mode of action as well as possible preventive measures.     

Sample preparation and high-resolution separation of mycotoxins possessing carboxyl groups

The chromatographic analysis of carboxyl-containing mycotoxins, such as fumonisin B₁, ochratoxin A, and citrinin, presents a continual challenge. Toxins must first be extracted from foods or tissues and then cleaned up before chromatographic separation and detection. Liquid–liquid extraction efficiencies for some carboxylic mycotoxins are marginal for spiked samples and uncertain for incurred residues. Immunoaffinity columns may be useful for concentrating mycotoxins from samples before chromatography. In almost every case, more than one analytical method must be used to confirm the identification of the mycotoxin. The fumonisins are especially troublesome to analyze because they are relatively insoluble in organic solvents, they are not separated easily by gas chromatography, and they do not respond to the usual absorbance or fluorescence detectors used in liquid chromatography. Fluorescence derivatization and electrospray liquid chromatography–mass spectrometry have now made it possible to detect trace levels of mycotoxins. The purity of mycotoxin standards for toxicological studies can be determined by liquid chromatography with either an evaporative light scattering detector or electrospray mass spectrometer. New developments in capillary electrophoresis, nonporous microsphere liquid chromatography, and detection methods for low-volatility compounds show promise for improving the analysis of mycotoxins in the future.     

Guidelines and considerations for the use of system suitability and quality control samples in mass spectrometry assays applied in untargeted clinical metabolomic studies

Background

Quality assurance (QA) and quality control (QC) are two quality management processes that are integral to the success of metabolomics including their application for the acquisition of high quality data in any high-throughput analytical chemistry laboratory. QA defines all the planned and systematic activities implemented before samples are collected, to provide confidence that a subsequent analytical process will fulfil predetermined requirements for quality. QC can be defined as the operational techniques and activities used to measure and report these quality requirements after data acquisition.

Aim of review

This tutorial review will guide the reader through the use of system suitability and QC samples, why these samples should be applied and how the quality of data can be reported.

Key scientific concepts of review

System suitability samples are applied to assess the operation and lack of contamination of the analytical platform prior to sample analysis. Isotopically-labelled internal standards are applied to assess system stability for each sample analysed. Pooled QC samples are applied to condition the analytical platform, perform intra-study reproducibility measurements (QC) and to correct mathematically for systematic errors. Standard reference materials and long-term reference QC samples are applied for inter-study and inter-laboratory assessment of data.

     

Ergosterol and Fusarium mycotoxins content in two maize cultivars under different forms of nitrogen fertilizers

Mycotoxins are secondary metabolites produced by many filamentous fungi, such as Fusarium, and are widespread in nature and can adversely affect almost all organisms. Mycotoxins can be formed in very different conditions such as when plants are growing or in crops that are stored in improper conditions. One of the methods to prevent the formation of mycotoxins in powdered feed is using varieties of appropriate quality and proper mineral fertilization, primarily nitrogen. In this study, the concentration of ergosterol and mycotoxins in the kernels of two maize cultivars from different maturity groups in Poland during the harvest of 2011 and 2012 was investigated using HPLC methods. We aimed to analyse the concentration of ergosterol as a fungal biomass indicator and mycotoxins from two maize cultivars under six different forms of nitrogen fertilizers. The “stay‐green” hybrid ES Paroli was characterized by a significantly lower ergosterol and the ability to accumulate the mycotoxins, compared to the classic cultivar ES Palazzo. The varieties of maize that remain green for a longer time period, the so‐called stay‐green type, are characterized by a different coefficient of nitrogen remobilization compared to the classic hybrids. Their dynamics of growth and accumulation of nutrients indicates a nitrogen fertilization system, indicating that slow‐release fertilizers are potentially more adapted to the cycles of maize vegetation. Hence, compared to the classic variant, the “stay‐green” variety uses nitrogen more effectively from mineral fertilizers. Such hybrids are healthier and remain in good condition for a longer time; therefore, they are characterized by a lower pathogen pressure. Thus, the cultivation of such varieties can be considered as one of the elements of integrated maize production. We observed identical reactions of the studied experimental factors for both vegetation seasons, which indicates the lack of environmental impact on the functioning and interaction of experimental factors.     

LIMS — beyond CSV: a strategy for LIMS data quality assurance

Computer system validation of a LIMS is often confused with the ongoing task of entering and quality-assuring the material, instrument, specification, method, calculation, and study data required for laboratory sample and results management. A practical approach to LIMS template data quality assurance includes preparation of one or more standard operating procedures for entry and quality assurance of material specifications, test limits, secondary calculation formulae, and other analytical laboratory testing information.     

Case study and application of process analytical technology (PAT) towards bioprocessing: Use of tryptophan fluorescence as at‐line tool for making pooling decisions for process chromatography

Process analytical technology (PAT) has been gaining momentum in the biopharmaceutical community due to the potential for continuous real time quality assurance resulting in improved operational control and compliance. Two imperatives for implementing any PAT tool are that “variability is managed by the process” and “product quality attributes can be accurately and reliably predicted over the design space established for materials used, process parameters, manufacturing, environmental, and other conditions.” Recently, we have been examining the feasibility of applying different analytical tools to bioprocessing unit operations. We have previously demonstarted that commercially available online‐high performance liquid chromatography and ultra performance liquid chromatography systems can be used for analysis that can facilitate real‐time decisions for column pooling based on product quality attributes (Rathore et al., 2008 a,b). In this article, we review an at‐line tool that can be used for pooling of process chromatography columns. We have demonstrated that our tryptophan fluorescence method offers a feasible approach and meets the requirements of a PAT application. It is significantly faster than the alternative of fractionation, offline analysis followed by pooling. Although the method as presented here is not an online method, this technique may offer better resolution for certain applications and may be a more optimal approach as it is very conducive to implementation in a manufacturing environment. This technique is also amenable to be used as an online tool via front face fluorescence measurements done concurrently with product concentration determination by UV. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Gaps in mammal conservation in China: An analysis with a framework based on minimum area requirements

Climate change, habitat loss, and human disturbance are major threats to biodiversity. Protecting habitats plays a pivotal role in biodiversity conservation, and there is a global imperative to establish an effective system of protected areas (PAs) to implement habitat conservation and halt biodiversity decline. However, the protected patch size of habitat for a species is just as important for biodiversity conservation as the expansion of areas already under protection. In China, conservation management is often carried out based on administrative divisions. Therefore, here, an analytical conservation management framework was developed based on administrative divisions to assess whether the current network of PAs can effectively meet species' conservation needs using the minimum area requirements (MARs) of species as criteria for medium and large-sized mammals in China. This study found that the MAR of medium and large-sized mammals was larger in the northwest and smaller in the southeast, while taking the Hu line as the dividing line. Precipitation seasonality, elevation, annual mean temperature, and annual precipitation are the main environmental factors driving the distribution of a species MAR. Compared with MAR for each species, the maximum protected patch size of habitat is severely undersized in most provinces where those species primarily distribute, and this is particularly true for large carnivores and threatened species. The densely populated provinces of eastern China are particularly affected by this. The present study's framework can identify the provinces needing to expand PAs or implement other effective area-based conservation measures and habitat restoration. This analytical framework is also relevant for biodiversity conservation in different taxa and regions around the globe.     

Recent advances in analysis of Chinese medical plants and traditional medicines

Chinese herbal medicine is gaining increasing popularity worldwide for health promotion and adjuvant therapy. Thus, selective and efficient analytical methods are required not only for quality assurance but also for authentication of the plant material. Applications of both chromatographic and electrophoretic techniques to the analysis of medicinal plants and Chinese traditional medicine preparations over the last 3 years are outlined in this review. The role of chemical fingerprinting is highlighted and a brief survey of determination of toxic components, natural and synthetic adulterants is also included. Moreover, different sample pretreatment and extraction methods are discussed.     

Effects of sand bar openings on some limnological variables in a hypertrophic tropical coastal lagoon of Brazil

This study describes the spatial and temporal dynamics of several physical, chemical and biological variables in the Grussai lagoon, and their relationship to ephemeral sand bar openings and to a constant in natura waste water input. The spatial variation in pH, dissolved oxygen, electrical conductivity, total alkalinity and nutrients (e.g. soluble reactive silicate, soluble reactive phosphate and ammonium) was associated to the anoxic and nutrient rich groundwater discharge, the development of aquatic macrophytes, the biological activities of phytoplanktonic community and the marine influence. During the period when the sand bar was closed (isolated), the lagoon water was supersaturated with dissolved oxygen and exhibited high values of pH (8–10), total alkalinity (3.000–5.000 μeq l^-1), and chlorophyll a contents (60-300 μg l^-1), and had low values of dissolved nutrients (nearly undetectable). These suggest a biological processes dominance. When the sand bar was opened, there was an enrichment with dissolved inorganic nutrients (e.g. ammonium and soluble reactive phosphorus up to 120 and 5 μM, respectively) and a decrease in pH (below 8), total alkalinity (below 3.000 μeq l^-1) and dissolved oxygen during the initial second to eight days. Subsequently there was a period when the physical and chemical characteristics of seawater prevailed. The lagoon returned chemical to the pre-opening water conditions in a few days (∼ 10–20). This quick return implies highly efficient biological mechanisms. The high levels of chlorophyll a, total nitrogen and phosphorus in the water column indicate a high eutrophication stage in the Grussai lagoon during the sand bar closed periods. This revised version was published online in July 2006 with corrections to the Cover Date.     

The New Quality Assurance Standard of the Joint Commission on Accreditation of Hospitals

The Joint Commission on Accreditation of Hospitals (JCAH) has continuously emphasized improvement in the quality of care provided in hospitals as the central purpose of the accreditation process. In striving to assure such improvement, the JCAH has stressed the need for, and the responsibility of, the medical and other professional staffs to provide continuing review and evaluation of patient care. In recent years, quality assessment activities have evolved, proliferated and matured to the extent that they require a purposeful integration if they are to effect sustained improvement in patient care and clinical performance. To assist hospitals in the coordination or integration of quality assessment activities, the Board of Commissioners of the JCAH has approved an important new quality assurance standard.Significant requirements of the new standard are a comprehensive quality assurance program, a written plan, a problem-focused approach to the review and evaluation of patient care and clinical performance, an annual reassessment of the program, and an improvement in patient care or clinical performance. The new standard reflects the JCAH belief that an integrated, problem-focused approach to quality assurance will significantly improve the quality of care provided throughout a hospital. Such an approach recognizes the interdependence of hospital departments and services in the provision of patient care, and, therefore, requires a purposeful integration or coordination of quality assessment data and activities. Consequently, quality assessment data may be utilized effectively and efficiently, and many potentially useless or duplicative quality assessment activities can be eliminated. The new standard affords hospitals considerable flexibility in the manner in which they implement and administer the program and encourages innovation.     

Quality assurance of biological monitoring in occupational and environmental medicine

Biological monitoring of chemical exposure in the workplace has become increasingly important in the assessment of health risk as an integral part of the overall occupational health and safety strategy. In environmental medicine biological monitoring plays also an important role in the assessment of excessive, acute or chronic exposure to chemical agents. To guarantee that the results obtained in biological monitoring are comparable with threshold limit values and results from other laboratories, the analysis must be carried out with tested and reliable analytical methods and accompanied by a quality assurance scheme. Confounding influences and interferences during the pre-analytical phase can be minimised by recommendations from experienced laboratories. For internal quality control commercially available control samples with an assigned concentration are used. External quality control programs for biological monitoring are offered by several institutions. The external quality control program of the German Society of Occupational and Environmental Medicine has been organised since 1982. In the meantime the 27th program has been carried out offering 96 analytes in urine, blood and plasma for 47 substances. This program covers most of the parameters relevant to occupational and environmental medicine. About 350 laboratories take part in these intercomparison programs. At present, ten German and 14 international laboratories are commissioned to determine the assigned values. The data evaluated from the results of the intercomparison programs give a good overview of the current quality of the determination of analytes assessed in occupational and environmental toxicological laboratories. For the analysis of inorganic substances in blood and urine the tolerable variation ranges from 7.5 to 43.5%. For organic substances in urine the tolerable variation ranges from 12 to 48%. The highest variations (36-60%) were found for the analysis of organochlorine compounds in plasma. The tolerable variations for the determination of solvents in blood by head space gas chromatography range from 26 to 57%. If the recommendations for the pre-analytical phase, the selection of reliable analytical methods by the laboratory and the carrying out of adequate quality control are observed, the pre-requisites for reliable findings during biological monitoring are fulfilled     

Panax quinquefolius: An overview of the contaminants

The root of Panax quinquefolius has been used as a food supplement and botanical drug throughout the world for many years. During growth, harvest, processing, and storage, it can be polluted by molds, pesticides, heavy metals, and other chemicals, resulting in contamination with mycotoxins, pesticide residues, and heavy metals, as well as other substances. These contaminants may be harmful to human health. In this review, we summarize the contaminants and significant advances that have been made in the study of these contaminants, focusing on the analytical methods and contamination management techniques for P. quinquefolius that have been developed over the last several decades.