A Study on the Mechanism and Properties of a Self-Powered H2O2 Electrochemical Sensor Based on a Fuel Cell Configuration with FePc and Graphene Cathode Catalyst Materials.

Conventional electrochemical sensors use voltammetric and amperometric methods with external power supply and modulation systems, which hinder the flexibility and application of the sensors. To avoid the use of an external power system and to minimize the number of electrochemical cell components, a self-powered electrochemical sensor (SPES) for hydrogen peroxide was investigated here. Iron phthalocyanine, an enzyme mimetic material, and Ni were used as a cathode catalyst and an anode material, respectively. The properties of the iron phthalocyanine catalyst modified by graphene nanoplatelets (GNPs) were investigated. Open circuit potential tests demonstrated the feasibility of this system. The GNP-modulated interface helped to solve the problems of aggregation and poor conductivity of iron phthalocyanine and allowed for the achievement of the best analytical characteristics of the self-powered H2O2 sensor with a low detection limit of 0.6 µM and significantly higher sensitivity of 0.198 A/(M·cm2) due to the enhanced electrochemical properties. The SPES demonstrated the best performance at pH 3.0 compared to pH 7.4 and 12.0. The sensor characteristics under the control of external variable load resistances are discussed and the cell showed the highest power density of 65.9 µW/cm2 with a 20 kOhm resistor. The practical applicability of this method was verified by the determination of H2O2 in blood serum.

Novel sample double dilution calibration method for determination of lithium in biological samples using automatic flow system with in-syringe reaction.

A novel sample double dilution calibration method (SDDCM) and an automatic flow system with in-syringe reaction and spectrophotometric detection were developed for determining lithium in biological samples. The method is based on the reaction of lithium with Thorin in an alkaline medium and the signal was measured at 480 nm. The reaction was performed simultaneously for both standards and samples in three syringes of the automatic flow system. The method was validated and successfully applied to the determination of lithium in synthetic and pharmaceutical samples, with results consistent with the ICP OES method. The novel calibration method, developed for the determination of lithium in biological samples, uses a sample with two dilution degrees. Using the method, the concentration of the analyte is determined by relating the signal for a less diluted sample to the calibration plot for a more diluted sample and vice versa. The implementation of the calibration method was facilitated by preparing solutions directly in the flow system. The use of two sample dilutions makes it possible to determine the analyte in the sample without preliminary preparation. Moreover, obtaining two results based on signals for a sample diluted to different degrees allows them to be verified for accuracy. The proposed approach was successfully verified by the determination of lithium in certified reference materials of blood serum and urine. Using the developed method lithium was determined within the concentration range of 0.06-1.5 mg L-1, with precision (CV, %) less than 6.7, and accuracy (RE, %) better than 6.9. The detection limit was 0.03 mg L-1.

DNA Aptamer Integrated Hydrogel Nanocomposites on Screen Printed Gold Electrodes for Point-of-Care Detection of Testosterone in Human Serum.

This work describes the development and evaluation of a novel electrochemical aptasensor for testosterone detection. The sensor utilizes a specifically designed DNA immobilized on a screen-printed gold electrode (SPGE) modified with a conductive hydrogel and gold nanoparticles (HG/NP) composite. The aptasensor exhibited a dose-dependent response to testosterone (0.05 to 50 ng/mL) with a detection limit of 0.14 ng/mL and a good sensitivity of 0.23 µA ng-1 mL cm-2. The sensor displayed excellent selectivity towards testosterone compared to structurally similar steroid hormones. Importantly, the incorporation of HG/NP not only improved the sensor's conductivity but also acted as an antifouling layer, minimizing signal interference from non-specific biomolecule interactions in complex biological samples like human serum. The results obtained from the aptasensor showed good correlation with a standard ELISA method, demonstrating its effectiveness in real-world scenarios.

Comparison of analytical approaches for the detection of oral testosterone undecanoate administration in men.

For antidoping laboratories, the determination of an illicit testosterone (T) administration in urine samples remains a difficult process as it requires the determination of the exogenous origin by carbon isotope ratios (CIRs) of testosterone and its metabolites. As a complement to the urinary analysis, targeting testosterone esters (e.g. testosterone undecanoate [TU]) in serum samples by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) could represent a simpler approach compared with isotope ratio mass spectrometry (IRMS). These two approaches both lead to the direct detection of the administration of exogenous T but with a difference in effort and complexity of the analysis. To compare the detection window obtained with the two strategies, serum and the corresponding urine samples collected from an administration study with oral TU were analysed. Results showed that, at all timepoints where the intact TU was detected in serum, the CIRs of urinary steroids were also not in agreement with an endogenous origin. IRMS analysis required more effort but resulted in slightly longer detection windows than the ester analysis. Finally, this comparison study showed that, in the presence of a suspicious urinary steroid profile, the LC-MS/MS steroid esters analysis in the corresponding serum samples can be very helpful. If steroid esters are not detected, the IRMS analysis can then be conducted on the urine sample afterwards. Overall, the combination of matrices might facilitate the detection of prohibited T administration in sports, especially for athletes with naturally low T/E ratios.

Advances in Sjögren's Syndrome Dry Eye Diagnostics: Biomarkers and Biomolecules beyond Clinical Symptoms.

Sjögren's syndrome dry eye (SSDE) is a subset of Sjögren's syndrome marked by dry eye symptoms that is distinct from non-Sjögren's syndrome dry eye (NSSDE). As SSDE can lead to severe complications, its early detection is imperative. However, the differentiation between SSDE and NSSDE remains challenging due to overlapping clinical manifestations. This review endeavors to give a concise overview of the classification, pathophysiology, clinical features and presentation, ocular and systemic complications, clinical diagnosis, and management of SSDE. Despite advancements, limitations in current diagnostic methods underscore the need for novel diagnostic modalities. Thus, the current review examines various diagnostic biomarkers utilized for SSDE identification, encompassing serum, salivary, and tear analyses. Recent advancements in proteomic research and exosomal biomarkers offer promising diagnostic potential. Through a comprehensive literature review spanning from 2016 to 2023, we highlight molecular insights and advanced diagnostic modalities that have the potential to enhance our understanding and diagnosis of SSDE.

Preparation of SERS base membrane with cellulose compound dopamine and determination of hypochlorite.

Flexible silver substrates were made by in situ reduction of silver nanoparticles in bacterial cellulose membranes using the unique advantage of dopamine. Subsequently, we modified the substrate with 4-mercaptophenol (4-MP), a molecule capable of specifically recognizing ClO-, and its corresponding SERS signal changes with the concentration of hypochlorite, thus allowing the quantitative detection of ClO- content. The method showed a negative linear correlation (R2 = 0.9567) with the SERS intensity at 1077 cm-1 over the concentration range 0.5-100 µM, and the detection limit was 0.15 µM. The RSD of the SERS intensity at 1077 cm-1 under five batches was 4.2%, which proved the good reproducibility of P-BCM-Ag NP-MP. Finally, the P-BCM-Ag NPs were used for the detection of hypochlorite in cell contents, artificial urine, and clinical serum samples, utilizing spike experiments in all three environments. The recoveries were in the range 90-110% indicating the accuracy of the method for the detection of hypochlorite and validating the promising application of this assay for practical detection in intricate biological samples.

Boron-cluster-based porous BCN material modified electrode for electrochemical determination of morphine in serum.

Porous highly boron-doped BCN (p-BCN) was produced by using a boron cluster salt (closo-[B12H12]2-) as the boron-based precursor and SiO2 as a hard template. The synthesized p-BCN was used in an electrochemical sensor for the ultrasensitive and highly selective detection of morphine (MOP). The optimal conditions for MOP detection were determined by optimizing the experimental conditions. Under these optimal conditions, the p-BCN-based sensor exhibited excellent MOP detection performance (working potential of 0.2 V). Specifically, it showed a detection range of 0.05 to 200 µM and a detection limit of 17.8 nM. Notably, the p-BCN-based electrochemical sensor was successfully applied to the determination of MOP in human blood, and the results showed satisfactory recovery and accuracy. Therefore, this sensor can be used as an effective platform for the detection of MOP in human blood samples.

Rapid determination of acyclovir, its main metabolite 9-carboxymethoxymethylguanine, ganciclovir, and penciclovir in human serum using LC-MS/MS.

A novel MS-based analytical method for simultaneous analysis of the antiviral drugs acyclovir, its metabolite 9-carboxymethoxymethylguanine, ganciclovir, and penciclovir in human serum is described. These antiviral drugs are active against herpes virus infections. Acyclovir and penciclovir are regarded as safe and effective medicines with mild side effects such as headache and gastrointestinal discomfort, and ganciclovir is regarded as more toxic and is known to cause, for example, bone marrow suppression. Acyclovir's main metabolite 9-carboxymethoxymethylguanine is a presumptive neurotoxin and should be monitored in patients with impaired renal function or in cases with neurotoxic symptoms. A sample was prepared using protein precipitation with 1% formic acid in methanol containing isotopically labeled internal standard. Chromatographic separation on a biphenyl column and mass spectrometric detection were performed in multiple reaction monitoring (MRM) mode on a Xevo TQ-S micro with ESI in positive ion mode, within 3 min. Inter-day assay accuracies for the quality controls varied between 95 and 104% and intra-day assay between 93 and 105%. Inter-day and intra-day assay imprecision for the quality controls ranged between 1.4 and 4.2% and 1.7 and 6.5% respectively. The lower limit of quantification for all four substances was 0.156 µmol/L. It is an accurate and reproducible method for therapeutic drug monitoring.

Fluorescent paper strip immunoassay with carbon nanodots@silica for determination of human serum amyloid A1.

A fluorescent paper strip immunoassay in conjunction with carbon nanodots@silica (CND@SiO2) as a label was developed for the quantitative measurements of human serum amyloid A1 (hSAA1) in serum at clinically significant concentrations for lung cancer diagnosis. Monodispersed CND@SiO2 was prepared by cohydrolysis between silane-crosslinked carbon nanodots and silica precursors via the Ströber method and further attached covalently to anti-hSAA1 (14F8) monoclonal antibody [anti-hSAA1(14F8)] specific to the hSAA1 target. The hSAA1 concentrations were then determined by quantifying the blue fluorescence intensity upon 365 nm excitation of the captured hSAA1 with anti-hSAA1(14F8)-CND@SiO2 conjugates in the test line on a paper strip where anti-hSAA1 (10G1) monoclonal antibody was physisorbed. The developed fluorescent paper strip with CND@SiO2 can detect hSAA1 at concentrations ranging from 0.1 to 5 nM (R2 = 0.995), with a limit of detection of  0.258 nM in 10 mM phosphate buffer pH 7.4 containing human serum albumin. The performance of  recovery (90.98-109.17%) and repeatability (coefficients of variation < 8.46%) obtained was also acceptable for quantitative determinations. The platform was employed for direct determination of hSAA1 concentrations in undiluted serum samples from lung cancer patients (relative standard deviation (RSD) < 7.46%) and healthy humans (RSD < 3.96%). The results were compared with those obtained using a commercially available enzyme-linked immunosorbent assay alongside liquid chromatography with tandem mass spectrometry measurements.

3D, large-area NiCo2O4 microflowers as a highly stable substrate for rapid and trace level detection of flutamide in biofluids via surface-enhanced Raman scattering (SERS).

A one-pot hydrothermal synthesis of three-dimensional (3D), large-area, bimetallic oxide NiCo2O4 (NCO) microflowers has been developed as a novel substrate for surface-enhanced Raman scattering (SERS) detection of flutamide in biological fluids. The 3D flower-like morphology of the NCO is observed via FESEM micrographs, while the orthorhombic phase formation is confirmed through XRD spectra. Due to the presence of multiple coordination cations of the 3D NCO microflowers (such as Ni2+ and Co2+), the high surface area and surface roughness, the NCO-modified indium tin oxide (NCO/ITO) SERS substrate exhibits a linear detection range from 0.5-500 nM with a low limit of detection (LOD) of 0.1 nM. The SERS substrate provides a high enhancement factor of 1.864 × 106 with an accumulation time of 30 s using a laser source of λ = 532 nm, which can be ascribed to the excellent and rapid interaction between the flutamide molecule and the NCO microflower substrate that leads to photoinduced charge transfer (PICT) resonance. The NCO/ITO substrate exhibits excellent homogeneity and high chemical stability. Besides, the substrate displays an excellent selectivity to flutamide molecules in the existence of other metabolites such as urea, ascorbic acid (AA), glucose, NaCl, KCl, CaCl2, and hydroxyflutamide. The NCO/ITO substrate is successful in the trace-level detection of flutamide in simulated blood serum samples. The strategy outlined here presents a novel strategy for the efficacy of transition metal oxides (TMOs) based electrodes useful for a wide variety of bioanalytical applications.

UHPLC-ESI-MS/MS Quantification of Relevant Substrates and Metabolites of the Kynurenine Pathway Present in Serum and Peritoneal Fluid from Gastric Cancer Patients-Method Development and Validation.

Metabolites and enzymes involved in the kynurenine pathway (KP) are highly promising targets for cancer treatment, including gastrointestinal tract diseases. Thus, accurate quantification of these compounds in body fluids becomes increasingly important. The aim of this study was the development and validation of the UHPLC-ESI-MS/MS methods for targeted quantification of biologically important KP substrates (tryptophan and nicotinamide) and metabolites(kynurenines) in samples of serum and peritoneal fluid from gastric cancer patients. The serum samples were simply pretreated with trichloroacetic acid to precipitate proteins. The peritoneal fluid was purified by solid-phase extraction before analysis. Validation was carried out for both matrices independently. Analysis of the samples from gastric cancer patients showed different accumulations of tryptophan and its metabolites in different biofluids of the same patient. The protocols will be used for the evaluation of tryptophan and kynurenines in blood and peritoneal fluid to determine correlation with the clinicopathological status of gastric cancer or the disease's prognosis.

Comparative Analysis of Serum Mineral and Biochemical Parameter Profiles Between Late Pregnant and Early Lactating Jennies.

The purpose of this study was to investigate the profile change of serum trace and major elements, and biochemical and hematological parameters in jennies during late pregnancy and early lactation. Twenty-five healthy Chinese Liaoxi jennies were used in late pregnancy and early lactation. Results showed that the levels of Fe, total protein (TP), and aspartate aminotransferase (AST) were highly variable interindividual among the jennies. Early lactating jennies showed significantly lower serum levels of K, Se, AST, total cholesterol (TC), and triglyceride than late pregnant jennies (P < .05). Principal component analysis identified six and five principal components of serum mineral and biochemical parameters for late pregnant and early lactating jennies, respectively, which was supported by the cluster analysis findings. Strong clustering of serum Cu-Mn, iPhos-Se-TP, and Ca-Zn-alanine aminotransferase-TC was found in the late pregnant jennies, and strong clustering of serum Ca-Zn-Se-Mn-albumin, Na-Fe-AST-triglyceride, and K-Mg-Cu-TP was observed in the early lactating jennies. The study suggests a significant variation in the serum levels of mineral and biochemical parameters in late pregnant and early lactating jennies, which is valuable in estimating their physiological status and providing proper health care.

Nickel sulfide nanoworm network architecture as a binder-free high-performance non-enzymatic glucose sensor.

Nickel sulfide nanoworm (Ni3S2 NW) network architecture was directly grown on the poly (3,4-ethylenedioxythiophene)-reduced graphene oxide hybrid films (PEDOT-rGO HFs) modified on glassy carbon electrode (GCE), acting as a binder-free sensor for high-performance non-enzymatic glucose monitoring. The sensor exhibited the satisfactory sensitivity (2123 µA mM-1 cm-2), wide linear range (15~9105 µM), low detection limit (0.48 µM), and rapid response time (< 1.5 s) at a potential of 0.5 V (vs. SCE) in 0.1 M NaOH and possessed good selectivity, reproducibility, and stability. The enhanced electrocatalytic activity of the sensor towards glucose oxidation was attributed to the particular morphology, satisfying hydrophilic nature, strong combination between Ni3S2 NWs, PEDOT-rGO, and bare GCE. Moreover, it can be used for assaying glucose in human serum samples without dilution, indicating potential for clinical diagnostic applications. Graphical abstract Nickel sulfide nanoworms (Ni3S2 NWs)/poly (3,4-ethylenedioxythiophene)-reduced graphene oxide hybrid films (PEDOT-rGO HFs) were used to construct a binder-free high-performance non-enzymatic glucose sensor with satisfactory sensitivity, wide linear range, low detection limit, good selectivity, amazing reproducibility, and stability.

Application of Graphdiyne in Surface-Assisted Laser Desorption Ionization Mass Spectrometry.

Graphdiyne (GD) is a new kind of carbon nanomaterial which has carbon-carbon triple bonds to form a layered structure. Here, we report the application of GD as the matrix for small molecule analysis in laser desorption ionization mass spectrometry (LDI MS). The GD matrix displayed two advantages: little background in the low mass range and good molecular ion signal in negative ion mode for many small molecules, e.g., fatty acids, amino acids, peptides, and drugs can be obtained in negative ion mode. By comparing the signal intensity of tetraphenylborate and juglone with and without GD existing, it was found that GD can enhance both of the desorption efficiency and ionization efficiency in LDI. Through analysis of the serum samples from liver cancer patients and healthy people, the GD-assisted LDI MS results showed that fatty acids could be used as potential biomarkers for the early diagnosis of liver cancer.

Base amount-dependent fluorescence enhancement for the assay of vascular endothelial growth factor 165 in human serum using hairpin DNA-silver nanoclusters and oxidized carbon nanoparticles.

A base amount-dependent fluorescence enhancement-based strategy is put forward to determine vascular endothelial growth factor 165 (VEGF165) in human serum by the use of hairpin DNA-silver nanoclusters (hDNA-AgNCs) and oxidized carbon nanoparticles (CNPs). The hDNA-AgNCs aptasensing probe consists of AgNCs-contained hairpin loop (that generates a fluorescence signal), hairpin stem (that makes the structure stable), and the terminal aptamer 1 (that recognizes the target together with aptamer 2). It has been demonstrated that the fluorescence intensity of hDNA-AgNCs is ~ 3-fold stronger than that of single-stranded DNA-AgNCs (ssDNA-AgNCs), and hDNA-AgNCs have a strong dependence of fluorescence enhancement on the base amount in hairpin stem and loop. Upon the addition of oxidized CNPs, the terminal aptamer 1 of hDNA-AgNCs can adsorb onto the surface of oxidized CNPs via π-π stacking, and the fluorescence of hDNA-AgNCs (with excitation/emission maxima at 490/567 nm) is quenched via fluorescence resonance energy transfer (FRET). When aptamer 2 and VEGF165 are subsequently added, aptamer 1, VEGF165, and aptamer 2 reassemble into an intact tertiary structure, and the fluorescence is recovered because hDNA-AgNCs are far away from the surface of oxidized CNPs and the FRET efficiency decreases. Under the optimized conditions, the aptasensing probe can selectively assay VEGF165 with a detection limit of 14 pM. The results provide a label-free and sensitive method to monitor VEGF165 in human serum. Schematic representation of the strong dependence of fluorescence enhancement on base amount in stem and loop of hairpin DNA-silver nanoclusters. The probe can be used to assay vascular endothelial growth factor 165 (VEGF165) and give a judgment whether human serum VEGF165 is at a normal or abnormal level for clinical diagnosis.

Advantages and Pitfalls of Capillary Electrophoresis of Pharmaceutical Compounds and Their Enantiomers in Complex Samples: Comparison of Hydrodynamically Opened and Closed Systems.

Several research disciplines require fast, reliable and highly automated determination of pharmaceutically active compounds and their enantiomers in complex biological matrices. To address some of the challenges of Capillary Electrophoresis (CE), such as low concentration sensitivity and performance degradation linked to the adsorption and interference of matrix components, CE in a hydrodynamically closed system was evaluated using the model compounds Pindolol and Propranolol. Some established validation parameters such as repeatability of injection efficiency, resolution and sensitivity were used to assess its performance, and it was found to be broadly identical to that of hydrodynamically opened systems. While some reduction in separation efficiency was observed, this was mainly due to dispersion caused by injection and it had no impact on the ability to resolve enantiomers of model compounds even when spiked into complex biological matrix such as blood serum. An approximately 18- to 23-fold increase in concentration sensitivity due to the employment of wide bore capillaries was observed. This brings the sensitivity of CE to a level similar to that of liquid chromatography techniques. In addition to this benefit and unlike in hydrodynamically opened systems, suppression of electroosmotic flow, which is essential for hydrodynamically closed systems practically eliminates the matrix effects that are linked to protein adsorption.

Electrochemiluminescence immunoassay of human chorionic gonadotropin using silver carbon quantum dots and functionalized polymer nanospheres.

A composite, reduced graphene oxide (rGO) doped with silver nanoparticles (Ag NPs), was prepared by using binary reductants of sodium citrate and hydrazine hydrate. Carbon quantum dots (CQDs) synthesized by papaya peel combined with silver ions to form a CQDs-loaded silver nanoparticle (AgCQDs) nanocomposite. Polymer nanospheres (PNS) were generated via the infinite coordination polymer of ferrocene dicarboxylic acid and employed as carriers to load AgCQDs. The prepared AgCQDs@PNS-PEI has good biocompatibility and electrical conductivity and can be used as a matrix for the immobilization of a secondary antibody (Ab2). A sandwich-type electrochemiluminescence (ECL) immunosensor using AgCQDs@PNS-PEI nanocomposite as probe has been developed for the detection of human chorionic gonadotropin (HCG). The proposed immunosensor exhibits a linear range from 0.00100 to 500 mIU mL-1 and the detection limit is 0.33 µIU mL-1 (S/N = 3) under optimal conditions. The sensor exhibits excellent selectivity, good reproducibility, and high stability. These features demonstrate that the proposed method has promising potential for clinical protein detection and displays a new strategy to fabricate an immunosensor. Graphical abstract.

Influence of IP-injected ZnO-nanoparticles in Catla catla fish: hematological and serological profile.

This study reports an effort to synthesize biocompatible zinc oxide nanoparticles using sol-gel method and its influence on hematological and serological profile of Catla catla fish. Hexagonal wurtzite structure and crystallite size of ZnO-NPs was identified by using XRD in the range of 19 to 20 nm. Moreover, the irregular and non-uniform surface of these NPs was found using SEM. The different stretched and vibrational mode (ZnO, OH, CO, and H-O-H) was identified by using FTIR analysis. UV-visible spectroscopy confirmed absorbance of the blue shift in the range 340 nm. Bioassay of ZnO-NPs on Catla catla was performed and nano ZnO was given through intraperitoneal injections at 0, 25, 50, 75, and 100 µg/g body weight doses. Analysis of fish blood samples indicated an increase in WBCs and leukocytes while the differential effect on monocytes. On the other hand in response to varying ZnO concentrations, an increase in RBCs, hemoglobin, and HCT was evident. Serum analysis revealed an increase in urea concentration while a reduction in creatinine, ALT, and AST. In an overall assessment, nano-ZnO supplementation at 25 to 100 µg/g body weight differentially affected hematological and serum biochemical profile of thaila fish. Graphical abstract.