Recent developments in the determination of urinary cancer biomarkers by capillary electrophoresis

Investigation of effective biomarkers for cancers is currently a popular area of study in clinical and cancer researches, because it can potentially lead to pre-cancer screening or pre-cancer diagnosis and may provide useful information on cancer type and the disease's stage of progression. More and more biochemical or chemical fluid components of the human body such as urine, blood, and cerebrospinal fluid have been considered to contain biomarkers, which are useful in cancer researches, pre-cancer diagnosis, and cancer follow-ups during or after cancer treatment. Several modern analytical techniques, such as gas chromatography (GC), high-performance liquid chromatography (HPLC), capillary electrophoresis (CE), and other separation techniques as well as hyphenated techniques, have been extensively used in study of cancer biomarkers. Among these techniques, CE is considered to be a highly efficient and practical analytical technique because of the small sample volume requirement and its wide separation versatility, ranging from small inorganic molecules to large biomolecules. This review discusses the latest developments involving biomarkers and their analysis by CE, including a discussion of instrumental conditions, method developments, and data analysis.     

Breath biosensing: using electrochemical enzymatic sensors for detection of biomarkers in human breath

Detection of biomarkers for disease by noninvasive methods is critical for the early diagnosis and screening of disease, enabling prompt treatment. Breath biosensors are a viable option as the exhaled breath contains several biomarkers linked to lung cancer, oxidative stress, diabetes, and other diseases. Breath analysis has been achieved by advanced analytical techniques such as gas chromatography and infrared spectroscopy. However, electrochemical enzymatic breath biosensors offer a cost-effective, sensitive platform for biomarker detection without complex analysis and interpretation by trained laboratory personnel. This review aims to summarize recent advances in the field of electrochemical enzymatic breath biosensors and offer future opportunities from other applications of nonelectrochemical enzymatic breath biosensors.     

Chromatographic and electrophoretic methods for the analysis of biomarkers of oxidative damage to macromolecules (DNA, lipids, and proteins)

Free radicals and other reactive species can cause oxidative damage to biomolecules when oxidant species exceed the antioxidant defences in the body, resulting in oxidative stress. Oxidatively damaged products have been associated with aging as well as with the development of pathologies like cancer, cardiovascular disease, neurodegenerative disorders, diabetes, inflammation, etc. Reliable measurements of biomarkers of oxidative damage to macromolecules would afford information on the pre-disposition and prognosis of certain pathologies, being of utmost importance in evaluation of the effect of intervention with antioxidants on the incidence of diseases associated to oxidative stress. This review will present and compare different analytical methods, especially those involving chromatographic and electrophoretic techniques, commonly used for the analysis of biomarkers of oxidative damage to the three main macromolecules, namely oxidised DNA, lipid peroxidation products, and protein carbonyls.     

Analysis of high-purity material—A comparison of photon activation analysis with other instrumental methods

Photon activation analysis (PAA) has been a well-established analytical method for about two decades. In this paper, its particular features are outlined in comparison with other modern instrumental techniques, especially radioanalytical methods, e.g. activation analysis with thermal or fast neutrons (NAA) or charged particles (CPA). Emphasis is placed upon the determination of trace components of high-purity material; iron, copper and lead matrix were selected as examples. In PAA, both instrumental and radiochemical procedures were applied. The results are presented in comparison with those obtained by other laboratories using a large variety of different techniques.     

Analyses of representative biomarkers of exposure and effect by chromatographic, mass spectrometric, and nuclear magnetic resonance techniques: method development and application in life sciences

Biomarkers are essential tools in monitoring studies, which include environmental monitoring, biological monitoring, biological effect monitoring, and health surveillance, as well as drug development processes. Their discovery, validation, and analysis require highly sensitive and selective analytical technologies. In this regard, gas and liquid chromatography, mass spectrometry, and nuclear magnetic resonance spectroscopy have facilitated great achievements in all these areas. In addition and closely related to biomarkers, the ongoing developments in these techniques promise a better understanding of the nature and mechanisms of toxic effects originating from various chemical, biological, or physical sources. This Review compiles studies performed on selected biomarkers with respect to both method development and application. Section 1 summarizes the concept of biomarkers; their application in various industrial/occupational, agricultural, drug developmental, and medical/clinical platforms. This section also focuses on biotransformation studies in close relation to biomarker discovery and validation, and on major techniques utilized in this area. In Section 2, biotransformation of volatile anesthetics in humans with a focus on mercapturic acid derivatives as potential biomarkers of effect is reviewed. The use of GC-ECD, GC/MS, and 19F-NMR in these studies is described. Section 3 focuses on the analysis of aldehydic lipid peroxidation degradation products by GC-ECD in mammalian cells in which oxidative stress induced chemically, and in humans after various challenges; anesthetic exposure, ischemia-reperfusion, and controlled endurance exercise. In Section 4, method development for protein and DNA oxidation products by LC-tandem MS and its application in mammalian cells and in humans are summarized. Possibilities, limitations, and future perspectives are discussed in Section 5.     

Ultrasensitive assays for proteins

Proteins are essential components of organisms and are involved in a wide range of biological functions. There are increasing demands for ultra-sensitive protein detection, because many important protein biomarkers are present at ultra-low levels, especially during the early stages of disease. Measuring proteins at low levels is also crucial for investigations of the protein synthesis and functions in biological systems. In this review, we summarize the recent developments of novel technology enabling ultrasensitive protein detection. We focus on two groups of techniques that involve either polymerase amplification of affinity DNA probes or signal amplification by the use of nano-/micro-materials. The polymerase-based amplification of affinity DNA probes indirectly improves the sensitivity of protein detection by increasing the number of detection molecules. The use of nano-/micro-materials conjugated to affinity probes enhances the measurement signals by using the unique electrical, optical, and catalytic properties of these novel materials. This review describes the basic principles, performances, applications, merits, and limitations of these techniques.     

Alkaline phosphatase labeled antibody-based electrochemical biosensor for sensitive HE4 tumor marker detection

Quantitative determination of the serum level of human epididymis protein 4 (HE4), a tumor marker for ovarian carcinoma, has come to the fore of interest mainly due to the possibility for its detection in early stages of the disease when the sensing of other biomarkers is limited. We present a simple ELISA based approach for rapid HE4 detection including its immunomagnetic capturing accompanied by sensitive electrochemical detection of the electroactive product formed after enzymatic conversion of the substrate by alkaline phosphatase used as a label of anti-HE4 IgG. The proposed immunosensor offers stability in time and due to excellent limit of detection at 6.8 fM HE4, and limit of quantification of 23 fM meets the requirements for the early detection of this biomarker.     

Concept of Selectivity and Its Meaning in Analytical Separation Techniques

The ambiguity of the use of selectivity concept in currently available analytical separation techniques (including chromatographic techniques) is pointed out. In all cases when this term is applied, it should be specified for which analyte (or analytes) and with respect to which concomitant component (or components) selectivity is considered in order to eliminate uncertainties of this kind. It should also be specified whether the analysis as a whole, its individual stages, chromatographic systems, or their particular elements (such as columns or phases used in them) are characterized. Corresponding criteria should also be specified for derived terms such as selectivity control, selectivity improvement, etc.     

Analysis of lipid peroxidation biomarkers in extremely low gestational age neonate urines by UPLC-MS/MS

Extremely low gestational age neonates (ELGAN) frequently require the use of oxygen supply in the delivery room leading to systemic inflammation and oxidative stress that are responsible for increased morbidity and mortality. The objective of this study was to establish reference ranges of a set of representative isoprostanes and prostaglandins, which are stable biomarkers of lipid peroxidation often correlated with oxidative stress-related disorders. First, a quantitative ultra performance liquid chromatography—tandem mass spectrometry (UPLC-MS/MS) method was developed and validated. The proposed analytical method was tailored for its application in the field of neonatology, enabling multi-analyte detection in non-invasive, small-volume urine samples. Then, the lipid peroxidation product concentrations in a total of 536 urine samples collected within the framework of two clinical trials including extremely low gestational age neonates (ELGAN) were analyzed. The access to a substantially large number of samples from this very vulnerable population provided the chance to establish reference ranges of the studied biomarkers. Up to the present, and for this population, this is the biggest reference data set reported in literature. Results obtained should assist researchers and pediatricians in interpreting test results in future studies involving isoprostanes and prostaglandins, and could help assessing morbidities and evaluate effectiveness of treatment strategies (e.g., different resuscitation conditions) in the neonatal field.

Applicability of bioanalysis of multiple analytes in drug discovery and development: review of select case studies including assay development considerations

The development of sound bioanalytical method(s) is of paramount importance during the process of drug discovery and development culminating in a marketing approval. Although the bioanalytical procedure(s) originally developed during the discovery stage may not necessarily be fit to support the drug development scenario, they may be suitably modified and validated, as deemed necessary. Several reviews have appeared over the years describing analytical approaches including various techniques, detection systems, automation tools that are available for an effective separation, enhanced selectivity and sensitivity for quantitation of many analytes. The intention of this review is to cover various key areas where analytical method development becomes necessary during different stages of drug discovery research and development process. The key areas covered in this article with relevant case studies include: (a) simultaneous assay for parent compound and metabolites that are purported to display pharmacological activity; (b) bioanalytical procedures for determination of multiple drugs in combating a disease; (c) analytical measurement of chirality aspects in the pharmacokinetics, metabolism and biotransformation investigations; (d) drug monitoring for therapeutic benefits and/or occupational hazard; (e) analysis of drugs from complex and/or less frequently used matrices; (f) analytical determination during in vitro experiments (metabolism and permeability related) and in situ intestinal perfusion experiments; (g) determination of a major metabolite as a surrogate for the parent molecule; (h) analytical approaches for universal determination of CYP450 probe substrates and metabolites; (i) analytical applicability to prodrug evaluations-simultaneous determination of prodrug, parent and metabolites; (j) quantitative determination of parent compound and/or phase II metabolite(s) via direct or indirect approaches; (k) applicability in analysis of multiple compounds in select disease areas and/or in clinically important drug-drug interaction studies. A tabular representation of select examples of analysis is provided covering areas of separation conditions, validation aspects and applicable conclusion. A limited discussion is provided on relevant aspects of the need for developing bioanalytical procedures for speedy drug discovery and development. Additionally, some key elements such as internal standard selection, likely issues of mass detection, matrix effect, chiral aspects etc. are provided for consideration during method development.     

Electrochemical Biosensing in Cancer Diagnostics and Follow‐up

In cancer, screening and early detection are critical for the success of the patient's treatment and to increase the survival rate. The development of analytical tools for non‐invasive detection, through the analysis of cancer biomarkers, is imperative for disease diagnosis, treatment and follow‐up. Tumour biomarkers refer to substances or processes that, in clinical settings, are indicative of the presence of cancer in the body. These biomarkers can be detected using biosensors, that, because of their fast, accurate and point of care applicability, are prominent alternatives to the traditional methods. Moreover, the constant innovations in the biosensing field improve the determination of normal and/or elevated levels of tumour biomarkers in patients’ biological fluids (such as serum, plasma, whole blood, urine, etc.). Although several biomarkers (DNA, RNA, proteins, cells) are known, the detection of proteins and circulating tumour cells (CTCs) are the most commonly reported due to their approval as tumour biomarkers by the specialized entities and commonly accepted for diagnosis by medical and clinical teams. Therefore, electrochemical immunosensors and cytosensors are vastly described in this review, because of their fast, simple and accurate detection, the low sample volumes required, and the excellent limits of detection obtained. The biosensing strategies reported for the six most commonly diagnosed cancers (lung, breast, colorectal, prostate, liver and stomach) are summarized and the distinct phases of the sensors’ constructions (surface modification, antibody immobilization, immunochemical interactions, detection approach) and applications are discussed.     

Capillary electrophoresis methods for microRNAs assays: A review

MicroRNAs (miRNAs) are short noncoding RNAs that conduct important roles in many cellular processes such as development, proliferation, differentiation, and apoptosis. In particular, circulating miRNAs have been proposed as biomarkers for cancer, diabetes, cardiovascular disease, and other illnesses. Therefore, determination of miRNA expression levels in various biofluids is important for the investigation of biological processes in health and disease and for discovering their potential as new biomarkers and drug targets. Capillary electrophoresis (CE) is emerging as a useful analytical tool for analyzing miRNA because of its simple sample preparation steps and efficient resolution of a diverse size range of compounds. In particular, CE with laser-induced fluorescence detection is a promising and relatively rapidly developing tool with the potential to provide high sensitivity and specificity in the analysis of miRNAs. This paper covers a short overview of the recent developments and applications of CE systems in miRNA studies in biological and biomedical areas.     

Umweltprobenbanken-eine wichtige Säule der Umweltbeobachtung

Natural events as well as human activities are affecting the environment and even with largescale monitoring programs it is not possible to completely assess the state of the environment. But a documentation of the environmental status with respect to its chemical composition can be realized with systematically collected representative environmental samples, which can be stored for decades at very low temperatures almost without degradation. Such ‘banking activity’ allows a current environmental monitoring and also a retrospective analysis for the determination of components which have not been analyzed at the time of sampling because of ta lack in analytical techniques or because they have not been known or considered environmentally relevant. The concept of environmental specimen banking and general procedures are described. Various examples illustrate the many facets of such activities.     

Isolation of neutral and acidic lipid biomarker classes for compound-specific-carbon isotope analysis by means of solvent extraction and normal-phase high-performance liquid chromatography

An analytical method for the quantitative determination of neutral and acidic lipid biomarkers in particulate and sediment samples has been developed. The method involves a first step with solvent extraction to isolate the neutral from the acidic compounds and a second step using normal-phase HPLC on a Nucleosil silica column to separate four different classes of neutral compounds: (1) aliphatic hydrocarbons, (2) aromatic hydrocarbons, (3) ketone compounds and (4) sterol and alcohol compounds. Recoveries of the individual spiked lipid biomarkers for the whole analytical procedure ranged from 88 to 106% for fatty acids, from 50 to 60% for aliphatic hydrocarbons (> or = n-C17), from 50 to 60% for polycyclic aromatic hydrocarbons (PAHs) (> or = 3 rings), 83% for friedelin and 60-80% for the sterol and alcohol compounds. The isolated compound classes were analysed by gas chromatography-combustion-isotope ratio mass spectrometry to measure the stable carbon isotope ratios in the individual compounds. The method enables the isolation of compound classes without fractionation for compound-specific carbon isotope analysis (delta13C). This analytical protocol has been applied, and proved suitable, for the determination of lipid biomarkers (sterols, fatty alcohols and fatty acids) in marine particulate material and for the determination of PAHs in sediment samples.     

醛类标志物的化学衍生化色谱-质谱分析方法进展

人体接触环境中的化学污染物会导致多种疾病,包括癌症、糖尿病、心血管疾病、神经退行性疾病(阿尔茨海默症、帕金森病等)等.作为一类具有高反应活性的亲电化合物,醛类(包括外源性醛类或环境污染物暴露后产生的内源性醛类)可与人体中多种重要生物分子形成共价修饰产物而产生毒害作用.暴露组研究自2005年被首次提出以来一直是一个前沿热...     

FT-IR analysis of single human normal and leukemic lymphocytes

The infrared spectra of single human normal and leukemic lymphocytes have been obtained by means of the microscope-FT-IR system. Substantial differences between the two kinds of cells have been observed at level of the O-P-O vibrations of the DNA and of the proteic components. This determination opens up the possibility in determining the early stages of leukemia, in following the course of patients under therapy, and in evaluation of the residual neoplastic disease.     

Liquid-phase microextraction of biomarkers: A review on current methods

The detection and quantification of biomarkers have gained more attention in the medical discipline to evaluating disease progression to manage medical treatment. Biomarkers range from gases to biological macromolecules. Because of the nanomolar range levels of typical biomarkers in plasma, blood, urine, exhalation samples, and other biological fluids as well as complex matrix of biological media, adequate sample preparation methods should be used for quantification of biomarkers. Biomarkers are discussed here generally classified mainly into two subgroups which arisen from disease or exposure compounds. The analytical method is critical for the validity/reliability of a biomarker. Accuracy, precision, reproducibility, recovery, sensitivity, and specificity all have high influence to the consistency with the limit and reference values concerned. In this paper, developments in well-established liquid-phase microextraction techniques for the clinical analysis of biological samples will be reviewed and discussed. This article presents an overview of microextraction methods for biological samples, focusing especially on biomarkers.     

Evolution of hyphenated techniques for mineral oil analysis in food

The occurrence of mineral oil in food is known since the early 1990s, and it was discovered by chance in one of the first applications using the hyphenated LC–GC system. Since then, the relationship between hyphenated techniques and mineral oil analysis has been tightly interrelated and successful. This review aims to show and explain how this mutual interaction has driven the development of the hyphenated techniques on one side and has supported the increase of knowledge on the other, supporting the complex task of mineral oil determination in food. The paper presents the background of the mineral oil problem in food (a brief history of its finding, toxicology, and occurrence), moving then toward the analytical determination. The development of different hyphenated techniques in relation to mineral oil determination is discussed, focusing mainly on 2D techniques, such as LC–GC. The necessity of additional dimensions, such as LC–LC–GC and comprehensive approaches, such as GC × GC and LC × GC, is also discussed. Finally, the role of the hyphenation with MS is presented.     

基于分子印迹与表面增强拉曼散射的蛋白质疾病标志物检测研究进展

异常的蛋白质表达与疾病的发生与发展密切相关,因此蛋白质已作为疾病标志物广泛应用于疾病的早期诊断、治疗监测和预后评估.然而,临床样本中的蛋白质疾病标志物通常含量极低,并存在高丰度的基质干扰,对检测方法的特异性和灵敏度提出挑战.目前,蛋白质疾病标志物的检测方法主要是免疫分析.但是,免疫分析主要依赖抗体进行特异性识别,而抗体...