Analysis of human biological samples using porous graphitic carbon columns and liquid chromatography-mass spectrometry: a review.

Liquid chromatography-mass spectrometry (LC-MS) has emerged as a powerful analytical technique for analyzing complex biological samples. Among various chromatographic stationary phases, porous graphitic carbon (PGC) columns have attracted significant attention due to their unique properties-such as the ability to separate both polar and non-polar compounds and their stability through all pH ranges and to high temperatures-besides the compatibility with LC-MS. This review discusses the applicability of PGC for SPE and separation in LC-MS-based analyses of human biological samples, highlighting the diverse applications of PGC-LC-MS in analyzing endogenous metabolites, pharmaceuticals, and biomarkers, such as glycans, proteins, oligosaccharides, sugar phosphates, and nucleotides. Additionally, the fundamental principles underlying PGC column chemistry and its advantages, challenges, and advances in method development are explored. This comprehensive review aims to provide researchers and practitioners with a valuable resource for understanding the capabilities and limitations of PGC columns in LC-MS-based analysis of human biological samples, thereby facilitating advancements in analytical methodologies and biomedical research.

Facile fabrication of a novel disposable pencil graphite electrode for simultaneous determination of promising immunosuppressant drugs mycophenolate mofetil and tacrolimus in human biological fluids.

An innovative electrochemical sensor was proposed for simultaneous determination of mycophenolate mofetil (Mph) and tacrolimus (TAC) for the first time. A novel sensor based on electro-polymerization of multi-walled carbon nanotubes (MWCNTs) and a novel Cu-1N-allyl-2-(2,5-dimethoxyphenyl)-4,5-diphenyl-1H-imidazole metal organic framework (Cu-ADPPI MOF) on disposable pencil graphite electrode (dPGE). Many techniques were used to characterize the electrochemical activity and surface structure of the fabricated sensor. The proposed sensor exhibited good catalytic performance towards Mph and TAC oxidation due to the synergistic effect. Under optimal conditions, the proposed sensor has achieved a linear range of 0.85-155 × 10-8 M and 1.1-170.0 × 10-8 M with LODs of 0.28 × 10-8 M and 0.36 × 10-8 M for Mph and TAC, respectively. The designated sensor showed good reproducibility, repeatability, stability, and selectivity for the determination of Mph and TAC. Moreover, the simultaneous determination of Mph and TAC in different human biological fluids was carried out with acceptable results. As a result, the proposed sensor opens a new venue for the use of electro-polymerized MOFs in combination with other conductive materials such as MWCNTs for electrochemical sensing of different analytes with the desired sensitivity and selectivity. Graphical abstract Construction of disposable graphite electrode, based on electro-deposition of multilayer films of multi-walled carbon nanotubes and a new generation of Cu-MOFs, for simultaneous analysis of tacrolimus and mycophenolate mofetil for the first time.

Organophosphorus pesticide determination in biological specimens: bioanalytical and toxicological aspects.

Organophosphorus insecticides, such as parathion-ethyl, quinalphos, chlorpyrifos, chlorfenvinphos or diazinon, are still widely used for pest control on crops. These compounds are extremely toxic to humans, and, even though specific legislation exists that controls the use of these substances, the frequency of toxic and/or fatal events and the existing data suggest that they are still easily accessed and the knowledge associated to the risks is not well-recognized. For these reasons, the determination of the exposure to these compounds, their detection (and of their metabolites as well) in biological samples, is of great importance in clinical and forensic toxicology, and, therefore, the development of techniques for this evaluation is an important task for laboratories. Most confirmatory analyses use blood, serum, plasma and urine as biological samples and are performed by either gas chromatographic-mass spectrometric or liquid chromatographic-mass spectrometric instrumentation, which represents the gold standard in what concerns high sensitivity. This paper will not only address the physical-chemical and toxicological aspects of this class of compounds but also perform a comprehensive and critical review on the analytical methods available for their determination in biological specimens, with special focus on the latest instrumental developments and sample preparation approaches.

Human biological sample biobanking to support tissue biomarkers in pharmaceutical research and development.

Advances in the understanding of molecular pathology and thereby the mechanisms that could be amenable to therapeutic manipulation are the reason that pharmaceutical research and development is focused increasingly on measurement of molecular biomarkers in human biological samples. Obtaining direct or indirect access to sufficient samples that are fit for research purposes can be a major challenge. A biobanking infrastructure has a significant role in the acquisition, storage and usage of human biological samples and here we review some key requirements for establishing a biobank. These include ensuring; that appropriate governance mechanisms are in place, that samples available are appropriate and fit for the intended research purposes that the infrastructure is sustainable in the future and that use of the biobank assets meets the strategic aims of the host organisation. Finally we present a case study--the STRATUM project which has recently completed and through a collaborative approach involving six industry and public partners drawing on a network of experts, examined biobank policies, public attitudes to biobanking, donor consent, sample and data standards, technical requirements for a register and biobanking financial models, albeit from a UK perspective.

Microplastic diagnostics in humans: "The 3Ps" Progress, problems, and prospects.

The growing global concern about human exposure to microplastics necessitates research into their occurrence, fate, and effects. Recent advancements in analytical methods have fostered research and improved understanding of microplastics in a variety of human tissue and biological samples, including blood, liver, lung, placenta, kidney, spleen, sputum, and feces, etc. Given the rapid expansion of this research topic, it is imperative to assess and introduce them to a broader audience. This article for the first time conducts a systematic review of the literature on microplastics in human biological samples, their objectives, current efforts, and key findings. This review offers an in-depth analysis of the research approaches employed, spanning from sampling to detection to quantification of microplastics, as well as an overview of their occurrence and characteristics to understand the level of microplastic exposure in the human body. It also provides a detailed analysis of existing contamination control procedures and attempts to build consistent cross-contamination prevention measures. Finally, we provide the reader with the guidelines on current microplastic research strategies, highlighting future directions. Overall, this synthesis will assist researchers in developing a multifaceted understanding of contemporary microplastic investigations in human biological samples.

A Phage Receptor-Binding Protein as a Promising Tool for the Detection of Escherichia coli in Human Specimens.

Escherichia coli is a problematic pathogen that causes life-threatening diseases, being a frequent causative agent of several nosocomial infections such as urinary tract and bloodstream infections. Proper and rapid bacterial identification is critical for allowing prompt and targeted antimicrobial therapy. (Bacterio)phage receptor-binding proteins (RBPs) display high specificity for bacterial surface epitopes and, therefore, are particularly attractive as biorecognition elements, potentially conferring high sensitivity and specificity in bacterial detection. In this study, we elucidated, for the first time, the potential of a recombinant RBP (Gp17) to recognize E. coli at different viability states, such as viable but not culturable cells, which are not detected by conventional techniques. Moreover, by using a diagnostic method in which we combined magnetic and spectrofluorimetric approaches, we demonstrated the ability of Gp17 to specifically detect E. coli in various human specimens (e.g., whole blood, feces, urine, and saliva) in about 1.5 h, without requiring complex sample processing.

Enzyme-free and label-free strategy for electrochemical oxaliplatin aptasensing by using rGO/MWCNTs loaded with AuPd nanoparticles as signal probes and electro-catalytic enhancers.

An innovative label free electrochemical aptasensor was developed for the analysis of oxaliplatin (OXAL) for the first time. The DNA oligonucleotide (aptamer) was successfully fabricated, by covalently attaching the amino terminus of the functional DNA on the glassy carbon electrode (GCE) surface modified with reduced graphene oxide (rGO) and multi-walled carbon nanotubes (MWCNTs) loaded with AuPd nanoparticles (AuPd NPs@rGO/MWCNTs/GCE). The stepwise assembly process of aptasensor on AuPd NPs@rGO/MWCNTs/GCE was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The aptamer-OXAL complex formation led to inhibition the electron transfer of Fe(CN)63-/4- on the electrode interface, which was clearly observed by decreasing the peak current of the redox probe. Furthermore, we managed to quantitatively measure OXAL by adding different concentrations of OXAL, while monitoring the decrease of differential pulse voltammogram (DPV) responses of the redox probe. Under the optimized conditions, the electrochemical aptasensor exhibited a linear range of 0.1-170.0 nmol L-1 with LOD of 60.0 pmol L-1. Next, we successfully applied the aptasensor calibrated system to determine OXAL in pharmaceutical injection and human biological samples.

Essential trace elemental levels (zinc, iron and copper) in the biological samples of smoker referent and pulmonary tuberculosis patients.

Tuberculosis is one of the major causes of illnesses and deaths throughout world particularly in Asia. Smoking is linked with tuberculosis recurrence and its mortality and may influence bacteriological conversion, clinical symptoms and treatment outcome. The aim of current study was to estimate association among essential trace elements {zinc (Zn), iron (Fe) and copper (Cu)} in human biological samples particularly blood, serum, scalp hair, saliva, sputum, and nasal fluid of smoking and nonsmoking pulmonary tuberculosis patients (n = 165, age ranged 16-35 years) residents of Hyderabad, Pakistan. The biological samples of age matched healthy controls were chosen as referents of both genders (n = 171) for the comparison purpose. The human biological samples were wet digested in microwave oven by 65 % HNO3 and 30 % H2O2 with (2:1) ratio. The concentrations of elements in acid digested samples were determined by atomic absorption spectrometry. The average zinc and iron concentration was lower, while level of copper was higher in the biological samples of pulmonary Tuberculosis patients as compared to referent subjects (p < .001). It was also concluded as a result of Zn and Fe deficiency combined with high contact of copper due to smoking of tobacco can be synergistic with the risk factors related with pulmonary tuberculosis.

On-line Ultrasound-Assisted Dispersive Micro-Solid-Phase Extraction Based on Amino Bimodal Mesoporous Silica Nanoparticles for the Preconcentration and Determination of Cadmium in Human Biological Samples.

On-line ultrasound-assisted dispersive micro-solid-phase extraction (USA-DµSPE) has been developed for preconcentration and separation of trace amounts of Cd(II) ions in 0.5 mL of human biological samples. In a syringe with a nylon membrane, new synthetic bulky amino bimodal mesoporous silica nanoparticles (NH2-UVM7) were dispersed as a nanoadsorbent in 5 mL of diluted serum sample (1:10), and after ultrasonic shaking, the liquid phase was separated from the solid phase. At the optimized pH, the chemical and physical adsorption of cadmium ions occurred, respectively, based on complexation with amine groups of UVM7 (Cd:NH2-UVM7) and silica nanoparticles. The analyte was then back-extracted from the sorbent with nitric acid solution (0.2 M), and its concentration was determined by electrothermal atomic absorption spectrometry (ETAAS). Under the optimized conditions, the linear range, limit of detection (LOD), and preconcentration factor (PF) were obtained as 0.01-0.56 µg L(-1), 0.002 µg L(-1), and 25, respectively. The adsorption capacity of NH2-UVM7 was found to be 108.6 mg g(-1) of cadmium. The validation of the methodology was performed by the human standard reference material (HSRM).

Biobanks and their importance in the clinical and scientific fields related to Spanish biomedical research.

The reality of biomedical research in Spain requires having an updated knowledge of the research reality and its ethical/legal framework. Research studies with human biological samples should be made with a sufficiently large number of samples to reflect the diversity of the human population, which meets the standard requirements to ensure optimum quality of the research results for further development. Furthermore, research with humans, and obtaining and/or deriving human biological samples and clinical research studies information is subject to a number of legal requirements and restrictions. Biobanks and biobank networks are established as the optimal structures that favor the storage of large volumes of human biological samples based on criteria to ensure their optimum quality, harmonization and security, respecting at all times, the ethical and legal requirements guaranteeing the rights of citizens.

Simultaneous analysis of antibiotics in biological samples by ultra high performance liquid chromatography-tandem mass spectrometry.

A rapid and reliable multiclass method was developed for the simultaneous analysis of 21 antibiotics (beta-lactams, aminoglycosides, penicillins, cephalosporins, carbapenems or quinolones) in urine, serum, cerebrospinal fluid (CSF) and bronchial aspirations by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Prior to chromatographic determination, the analytes were extracted from human biological fluids by simple sample treatments, which imply dilution, liquefaction, or protein precipitation. Several chromatographic conditions were optimized in order to obtain a fast separation (<6min for each chromatographic run). MS/MS conditions were evaluated in order to increase selectivity and sensitivity and all compounds were detected in electrospray (ESI) positive ion mode, except clavulanic acid and sulbactam, which were monitored in negative ion mode. The developed method was validated in terms of linearity, selectivity, limits of detection (LODs) and quantification (LOQs), trueness, repeatability and interday precision. The LOQs ranged from 0.01 to 1.00mg/L for urine, serum and CSF. In case of bronchial aspirations, the LOQs were between 0.02 and 0.67mg/kg. In all matrices the recovery results were in the range 70-120% and interday precision was lower than 25%. Finally, the optimized method was applied to the analysis of biological samples from 10 patients in the intensive care unit (ICU) of a hospital located in Almeria (Spain). Several antibiotics (e.g., amoxicillin, tobramycin, levofloxacin, or linezolid) were found in the studied samples, observing that the highest concentrations were obtained in urine samples.