Harmonisation of Measurement Procedures: how do we get it done?

Clinical laboratory measurement results must be comparable among different measurement procedures, different locations and different times in order to be used appropriately for identifying and managing disease conditions. Harmonisation in the broad sense is the overall process of achieving comparability of results among clinical laboratory measurement procedures that measure the same measurand. The term standardisation is used when comparable results among measurement procedures are based on calibration traceability to SI using a reference measurement procedure of the highest available order. When there is no higher order reference measurement procedure available, and it is unlikely that one can be developed, the term harmonisation refers to any process for achieving comparable results among measurement procedures for an individual measurand.This review explains calibration traceability and focuses on the principles of harmonisation for those measurands for which a reference measurement procedure does not exist. We discuss the value of harmonisation, the importance of commutable reference materials, the barriers to harmonisation that exist today, and conclude with a discussion of a current global effort to improve the state of harmonisation.     

Reference materials and reference measurement procedures: an overview from a national metrology institute

An outline of the processes involved in both certified clinical reference material production and clinical reference measurement procedure development at the National Institute of Standards and Technology (NIST), the national metrology institute of the United States, is presented. The role that NIST and other national metrology institutes play in the metrological traceability of certified reference material is discussed. Highlighted are the challenges associated with the development of reference measurement systems for complex clinical analytes, such as proteins, and examples of existing efforts in this area are given. Examples of recent international collaborations in developing certified reference materials for analytes such as cardiac troponin I, brain natriuretic peptide, and serum creatinine demonstrate the close cooperation that national metrology institutes must have with the clinical community to establish complete reference measurement systems.     

Strain,biochemistry, and cultivation-dependent measurement variability of algal biomass composition

Accurate compositional analysis in biofuel feedstocks is imperative; the yields of individual components can define the economics of an entire process. In the nascent industry of algal biofuels and bioproducts, analytical methods that have been deemed acceptable for decades are suddenly critical for commercialization. We tackled the question of how the strain and biochemical makeup of algal cells affect chemical measurements. We selected a set of six procedures (two each for lipids, protein, and carbohydrates): three rapid fingerprinting methods and three advanced chromatography-based methods. All methods were used to measure the composition of 100 samples from three strains: Scenedesmus sp., Chlorella sp., and Nannochloropsis sp. The data presented point not only to species-specific discrepancies but also to cell biochemistry-related discrepancies. There are cases where two respective methods agree but the differences are often significant with over- or underestimation of up to 90%, likely due to chemical interferences with the rapid spectrophotometric measurements. We provide background on the chemistry of interfering reactions for the fingerprinting methods and conclude that for accurate compositional analysis of algae and process and mass balance closure, emphasis should be placed on unambiguous characterization using methods where individual components are measured independently.     

Ion mobility–mass spectrometry for structural proteomics

Ion mobility coupled to mass spectrometry has been an important tool in the fields of chemical physics and analytical chemistry for decades, but its potential for interrogating the structure of proteins and multiprotein complexes has only recently begun to be realized. Today, ion mobility–mass spectrometry is often applied to the structural elucidation of protein assemblies that have failed high-throughput crystallization or NMR spectroscopy screens. Here, we highlight the technology, approaches and data that have led to this dramatic shift in use, including emerging trends such as the integration of ion mobility–mass spectrometry data with more classical (e.g., ‘bottom-up’) proteomics approaches for the rapid structural characterization of protein networks.     

Metrology for ultrasonic applications

This paper provides a review of current metrological capability applied to the characterisation of the acoustic output of equipment used within medical ultrasonic applications. Key measurement devices, developed to underpin metrology in this area, are the radiation force balance, used to determine total output power, and the piezo-electric hydrophone, used to resolve the spatial and temporal distribution of acoustic pressure. The measurement infrastructure in place within the United Kingdom ensuring users are able to carry out traceable measurements of these quantities in a meaningful way, is described. This includes the relevant primary standards, the way international equivalence of national standards is demonstrated and the routes by which the standards are disseminated to the user community. Emerging measurement techniques that may in future lead to improved measurement capability, are also briefly discussed.     

Quantitative determination of cellulase concentration as distinct from cell concentration in studies of microbial cellulose utilization: analytical framework and methodological approach.

In analyzing microbial cellulose utilization, it would be useful to independently measure the mass concentration of cells and cellulase enzymes. Such measurements would allow investigation of the allocation of cellular resources between synthesis of cells and cellulase, in vivo cell- and cellulase-specific cellulose hydrolysis rates, and bioenergetics. Methodological protocols are not established for independent determination of cell and cellulase concentrations for the common case in which a substantial fraction of cellulase is attached to the cell surface. Alternative analytical approaches by which to develop such protocols are examined from the perspective of error minimization. For cell concentration measurement, acceptable accuracy is expected when the concentrations of a cell-specific component (e.g., DNA) is determined or when total protein is determined in conjunction with a measurement specific to cellulase. For cellulase concentration measurement, acceptable accuracy is expected when a measurement specific to cellulase such as ELISA is used. Several analytical approaches are rejected based on large expected errors.     

Measuring cortisol,the major stress hormone in fishes

Stress in teleosts is an increasingly studied topic because of its interaction with growth, reproduction, immune system and ultimately fitness of the animal. Whether it is for evaluating welfare in aquaculture, adaptive capacities in fish ecology, or to investigate effects of human-induced rapid environmental change, new experimental methods to describe stress physiology in captive or wild fish have flourished. Cortisol has proven to be a reliable indicator of stress and is considered the major stress hormone. Initially principally measured in blood, cortisol measurement methods are now evolving towards lower invasiveness and to allow repeated measurements over time. We present an overview of recent achievements in the field of cortisol measurement in fishes, discussing new alternatives to blood, whole body and eggs as matrices for cortisol measurement, notably mucus, faeces, water, scales and fins. In parallel, new analytical tools are being developed to increase specificity, sensitivity and automation of the measure. The review provides the founding principles of these techniques and introduces their potential as continuous monitoring tools. Finally, we consider promising avenues of research that could be prioritised in the field of stress physiology of fishes.     

Matrix solid phase dispersion (MSPD)

A review of the many uses of matrix solid phase dispersion (MSPD) in the extraction and analysis of a variety of compounds from a range of samples is provided. Matrix solid phase dispersion (MSPD) has found particular application as a somewhat generic analytical process for the preparation, extraction and fractionation of solid, semi-solid and/or highly viscous biological samples. Its simplicity and flexibility contribute to it being chosen over more classical methods for these purposes. MSPD is based on several simple principles of chemistry and physics, involving forces applied to the sample by mechanical blending to produce complete sample disruption and the interactions of the sample matrix with a solid support bonded-phase (SPE) or the surface chemistry of other solid support materials. These principles are discussed as are the factors to be considered in conducting a MSPD extraction.     

Matrix solid phase dispersion (MSPD)

A review of the many uses of matrix solid phase dispersion (MSPD) in the extraction and analysis of a variety of compounds from a range of samples is provided. Matrix solid phase dispersion (MSPD) has found particular application as a somewhat generic analytical process for the preparation, extraction and fractionation of solid, semi-solid and/or highly viscous biological samples. Its simplicity and flexibility contribute to it being chosen over more classical methods for these purposes. MSPD is based on several simple principles of chemistry and physics, involving forces applied to the sample by mechanical blending to produce complete sample disruption and the interactions of the sample matrix with a solid support bonded-phase (SPE) or the surface chemistry of other solid support materials. These principles are discussed as are the factors to be considered in conducting a MSPD extraction.     

Methods of quantitative analysis of the nitric oxide metabolites nitrite and nitrate in human biological fluids

In human organism, the gaseous radical molecule nitric oxide (NO) is produced in various cells from L-arginine by the catalytic action of NO synthases (NOS). The metabolic fate of NO includes oxidation to nitrate by oxyhaemoglobin in red blood cells and autoxidation in haemoglobin-free media to nitrite. Nitrate and nitrite circulate in blood and are excreted in urine. The concentration of these NO metabolites in the circulation and in the urine can be used to measure NO synthesis in vivo under standardized low-nitrate diet. Circulating nitrite reflects constitutive endothelial NOS activity, whereas excretory nitrate indicates systemic NO production. Today, nitrite and nitrate can be measured in plasma, serum and urine of humans by various analytical methods based on different analytical principles, such as colorimetry, spectrophotometry, fluorescence, chemiluminescence, gas and liquid chromatography, electrophoresis and mass spectrometry. The aim of the present article is to give an overview of the most significant currently used quantitative methods of analysis of nitrite and nitrate in human biological fluids, namely plasma and urine. With minor exception, measurement of nitrite and nitrate by these methods requires method-dependent chemical conversion of these anions. Therefore, the underlying mechanisms and principles of these methods are also discussed. Despite the chemical simplicity of nitrite and nitrate, accurate and interference-free quantification of nitrite and nitrate in biological fluids as indicators of NO synthesis may be difficult. Thus, problems associated with dietary and laboratory ubiquity of these anions and other preanalytical and analytical factors are addressed. Eventually, the important issue of quality control, the use of commercially available assay kits, and the value of the mass spectrometry methodology in this area are outlined.     

Synthetic biology: lessons from the history of synthetic organic chemistry

The mid-nineteenth century saw the development of a radical new direction in chemistry: instead of simply analyzing existing molecules, chemists began to synthesize them--including molecules that did not exist in nature. The combination of this new synthetic approach with more traditional analytical approaches revolutionized chemistry, leading to a deep understanding of the fundamental principles of chemical structure and reactivity and to the emergence of the modern pharmaceutical and chemical industries. The history of synthetic chemistry offers a possible roadmap for the development and impact of synthetic biology, a nascent field in which the goal is to build novel biological systems.     

The inverse problems for some topological indices in combinatorial chemistry.

In the original paper, Goldman et al. (2000) launched the study of the inverse problems in combinatorial chemistry, which is closely related to the design of combinatorial libraries for drug discovery. Following their ideas, we investigate four other topological indices, i.e., the sigma-index, the c-index, the Z-index, and the M(1)-index, with a special emphasis on the sigma-index. Like the Wiener index, these four indices are very popular in combinatorial chemistry and reflect many chemical and physical properties. We give algorithmic and analytical solutions for the inverse problems of the four indices. We also show that the SUBTREEVALUE reconstruction problem for the sigma-index is NP-hard.     

ReviewMethods of quantitative analysis of the nitric oxide metabolites nitrite and nitrate in human biological fluids

In human organism, the gaseous radical molecule nitric oxide (NO) is produced in various cells from l-arginine by the catalytic action of NO synthases (NOS). The metabolic fate of NO includes oxidation to nitrate by oxyhaemoglobin in red blood cells and autoxidation in haemoglobin-free media to nitrite. Nitrate and nitrite circulate in blood and are excreted in urine. The concentration of these NO metabolites in the circulation and in the urine can be used to measure NO synthesis in vivo under standardized low-nitrate diet. Circulating nitrite reflects consitutive endothelial NOS activity, whereas excretory nitrate indicates systemic NO production. Today, nitrite and nitrate can be measured in plasma, serum and urine of humans by various analytical methods based on different analytical principles, such as colorimetry, spectrophotometry, fluorescence, chemiluminescence, gas and liquid chromatography, electrophoresis and mass spectrometry. The aim of the present article is to give an overview of the most significant currently used quantitative methods of analysis of nitrite and nitrate in human biological fluids, namely plasma and urine. With minor exception, measurement of nitrite and nitrate by these methods requires method-dependent chemical conversion of these anions. Therefore, the underlying mechanisms and principles of these methods are also discussed. Despite the chemical simplicity of nitrite and nitrate, accurate and interference-free quantification of nitrite and nitrate in biological fluids as indicators of NO synthesis may be difficult. Thus, problems associated with dietary and laboratory ubiquity of these anions and other preanalytical and analytical factors are addressed. Eventually, the important issue of quality control, the use of commercially available assay kits, and the value of the mass spectrometry methodology in this area are outlined.     

An analytical perspective on detection,screening, and confirmation in phycology,with particular reference to toxins and toxin‐producing species

Knowledge concerning the ability of microalgae to produce metabolites of interest such as toxins or high‐value secondary metabolites requires exhaustive details to be supplied on how the research was conducted. These should include the microalgal species and strain characterization, the culture conditions, the cell density, and physiological state at the time of harvesting, the harvesting method, the sample pre‐treatment protocol, and the subsequent instrumental analytical separation/detection system. In this comment, we discuss issues that affect algal research from an analytical chemistry perspective, particularly (i) the need to specify detection capabilities of the entire method (i.e., limits of detection or threshold detection levels), which we illustrate in relation to classification of a species or strain as being “toxin producing” or “non‐toxin producing”; and (ii) the requirements that have to be satisfied to confirm a microalgal species (new or not) as a producer of a particular chemical of interest for phycologists, which again we illustrate in relation to toxins. A successful collaboration among phycologists and analytical chemists will only be achieved as a result of a synergistic collaboration between the two disciplines, with a reciprocal understanding at least at a background level.     

Uncertainty of Measurement: A Review of the Rules for Calculating Uncertainty Components through Functional Relationships

The Evaluation of Measurement Data - Guide to the Expression of Uncertainty in Measurement (usually referred to as the GUM) provides general rules for evaluating and expressing uncertainty in measurement. When a measurand, y, is calculated from other measurements through a functional relationship, uncertainties in the input variables will propagate through the calculation to an uncertainty in the output y. The manner in which such uncertainties are propagated through a functional relationship provides much of the mathematical challenge to fully understanding the GUM.The aim of this review is to provide a general overview of the GUM and to show how the calculation of uncertainty in the measurand may be achieved through a functional relationship. That is, starting with the general equation for combining uncertainty components as outlined in the GUM, we show how this general equation can be applied to various functional relationships in order to derive a combined standard uncertainty for the output value of the particular function (the measurand). The GUM equation may be applied to any mathematical form or functional relationship (the starting point for laboratory calculations) and describes the propagation of uncertainty from the input variable(s) to the output value of the function (the end point or outcome of the laboratory calculation). A rule-based approach is suggested with a number of the more common rules tabulated for the routine calculation of measurement uncertainty.     

Searching for life in the Universe: unconventional methods for an unconventional problem

The search for life, on and off our planet, can be done by conventional methods with which we are all familiar. These methods are sensitive and specific, and are often capable of detecting even single cells. However, if the search broadens to include life that may be different (even subtly different) in composition, the methods and even the approach must be altered. Here we discuss the development of what we call non-earthcentric life detection – detecting life with methods that could detect life no matter what its form or composition. To develop these methods, we simply ask, can we define life in terms of its general properties and particularly those that can be measured and quantified? Taking such an approach we can search for life using physics and chemistry to ask questions about structure, chemical composition, thermodynamics, and kinetics. Structural complexity can be searched for using computer algorithms that recognize complex structures. Once identified, these structures can be examined for a variety of chemical traits, including elemental composition, chirality, and complex chemistry. A second approach involves defining our environment in terms of energy sources (i.e., reductants), and oxidants (e.g. what is available to eat and breathe), and then looking for areas in which such phenomena are inexplicably out of chemical equilibrium. These disequilibria, when found, can then be examined in detail for the presence of the structural and chemical complexity that presumably characterizes any living systems. By this approach, we move the search for life to one that should facilitate the detection of any earthly life it encountered, as well as any non-conventional life forms that have structure, complex chemistry, and live via some form of redox chemistry. Electronic Publication