Electrochemical Monitoring of Methotrexate Anticancer Drug in Human Blood Serum by Using in situ Solvothermal Synthesized Fe3O4/ITO Electrode

Here, we reported on a one‐step fabrication of magnetite Fe₃O₄ nanoparticles/indium tin oxide (ITO) electrode based on the direct growing of Fe₃O₄ nanoparticles on the ITO surface by using a solvothermal process. The modified electrode was used as electrochemical methotrexate (MTX) biosensor with high sensitivity based on cyclic voltammetry and square wave voltammetry techniques. The results demonstrated a linear relationship between the MTX concentration and its oxidation current peak over a wide range from 10^?5 to 10^?14 mole/L with a limit of detection of 0.4×10^?15 M based on the square wave voltammetry (SWV) technique. In addition, Fe₃O₄/ITO electrode showed a good capability for measuring very low concentrations of MTX drug dissolved in human serum solution. Also, Fe₃O₄/ITO electrode was used for detecting MTX in blood serum samples collected from patients after their treatment with MTX. The prepared electrode showed the higher sensitivity that higher than the Viva‐E instrument, which opens the door for developing a cheap, simple and higher sensitive MTX sensor.     

Quantification of retinoid concentrations in human serum and brain tumor tissues

Retinoic acid signaling is essential for central nervous system (CNS) differentiation and appears to be impaired in tumors. Thus far, there are no established methods to quantify relevant retinoids (all-trans-retinoic acid, 9-cis-retinoic acid, 13-cis retinoic acid, and retinol) in human brain tumors. We developed a single step extraction and quantification procedure for polar and apolar retinoids in normal tissue, lipid-rich brain tumor tissues, and serum. This quantification procedure is based on high performance liquid chromatography (HPLC) with diode-array detection (DAD) using all-trans-acitretin as an internal standard and extraction by liquid–liquid partition with ethyl acetate and borate buffer at pH 9. Recovery with this extraction procedure was higher than earlier (two-step) liquid–liquid extraction procedures based on hexane, NaOH, and HCl. The overall quantification procedure was validated according to Food and Drug Administration (FDA) guidelines and fulfilled all criteria of accuracy, precision, selectivity, recovery, and stability. The overall method accuracy varied between −5.6% and +5.4% for serum and −3.8% and +6.2% for tissues, and overall precision ranged from 3.1% to 6.9% for serum and 2.1% to 8.3% for tissues (%CV batch-to-batch). The lower limit of quantification for all compounds in tumor tissue (and serum) was 3.9 ng g⁻¹ (ng mL⁻¹). Using this assay, photodegradation of the retinoids was evaluated and endogenous polar and apolar retinoids were quantified in sera and brain tumor tissues of patients and compared with serum and tonsil tissue concentrations of controls. It may thus serve as a suitable method for the characterization of retinoid uptake and metabolism in the respective compartments.     

A novel sorptive extraction method based on polydimethylsiloxane frit for determination of lung cancer biomarkers in human serum

In this study, a porous polypropylene frit was coated with polydimethylsiloxane (PDMS) as extraction medium, based on the home-made PDMS-frit, a rapid, simple and sensitive sorptive extraction method was established for analysis of potential biomarkers of lung cancer (hexanal and heptanal) in human serum samples. In the method, derivatization and extraction occurred simultaneously on the PDMS-frit, then the loaded frit was ultrasonically desorbed in acetonitrile. Polymerization, derivatization–extraction and desorption conditions were optimized. Under the optimal conditions, satisfactory results were gained, a wide linear application range was obtained in the range of 0.002–5.0 μmol L⁻¹ (R > 0.997) for two aldehydes, the detection limits (S N⁻¹ = 3) were 0.5 nmol L⁻¹ for hexanal and 0.4 nmol L⁻¹ for heptanal. The relative standard deviations (RSDs, n = 5) of the method were below 7.9% and the recoveries were above 72.7% for the spiked serum. All these results hint that the proposed method is potential for disease markers analysis in complex biological samples.     

Iron Interference in the Measurement of Selenium in Whole Blood,Plasma and Serum by Graphite Furnace Atomic Absorption

Abstract

Selenium is considered to be a trace element and the determination of diagnostic levels are most conveniently measured in blood, plasma or serum in humans and animals. The approach that is taken varies according to laboratory preferences. One such method involves the use of Graphite furance Atomic Absorption Spectroscopy (GFAAS). The simplicity of diluting the sample followed by direct determination without further preparation is the most attractive characteristic of this method, coupled with good sensitivity. Proper precautions must be taken however, including that of choosing the proper instrumental parameters for the analysis. If deuterium background correctiopn is used then then analyses should be carried out at the 204.0 nm line of Selenium. By using a matrix modifier consisting of Cu/Mg, deuterium background correction and off the wall of the graphite tube atomization, the determination of the analyte was demonstrated to be possible at the mentioned wavelength, free from iron interference present in the samples.     

Immunosensor for the Measurement of Human Serum Albumin in Urine Based on the Spreeta Surface Plasmon Resonance Sensor

The application of SPR for measurement of the concentration of human serum albumin (HSA) in urine was studied using the compact integrated SPR sensing system Spreeta. HSA was immobilized via cystamine and glutaraldehyde onto the gold sensing area and a competitive assay for HSA was developed using a limited amount of the monoclonal antibody AL-01 in solution. Measurements were carried out in the flow-through mode and the interaction between immobilized HSA and antibody was observed in real time. To obtain reproducible results, different conditions of the measurement (method of immobilization of HSA, data evaluation, concentration of antibody, regeneration procedure) were tested. The calibration curve for clinically relevant concentrations of HSA in urine samples was constructed using 300-times diluted antibody in the form of ascites fluid. The measuring range was between 0.1 and 5 mg/l of HSA, the sensing surface was successfully regenerated and suitable for more than 20 assays. The developed method was tested on real samples of urine; to overcome the non-specific adsorption of urine components, the differential approach was adopted and the measured signal was corrected by subtraction of the response observed in the absence of the antibody.     

Concurrent extraction,clean‐up,and analysis of polybrominated diphenyl ethers,hexabromocyclododecane isomers,and tetrabromobisphenol A in human milk and serum

A method has been developed and validated for the concurrent extraction, clean‐up, and analysis of polybrominated diphenyl ethers (PBDEs), α‐, β‐, and γ‐hexabromocyclododecane (HBCD), and tetrabromobisphenol A (TBBPA) in human milk and serum. Milk and serum samples were extracted using accelerated solvent extraction with acetone/hexane 1:1, v/v and liquid–liquid extraction with methyl‐tert‐butyl ether/hexane 1:1, v/v, respectively. The removal of co‐extracted biogenic materials was achieved by gel permeation chromatography followed by sulfuric acid treatment. The fractionation of the PBDEs and HBCD/TBBPA was performed using a Supelco LC‐Si SPE cartridge. The detection of the PBDEs was then performed by GC–MS and that of the HBCDs and the TBBPA was performed using UPLC–MS/MS. The pretreatment procedure was optimized, and the characteristic ions and fragmentation of the analytes were studied by MS or MS/MS. A recovery test was performed using a matrix spiking test at concentrations of 0.05–10 ng/g. The recoveries ranged from 78.6–108.8% with RSDs equal to or lower than 14.04%. The LODs were 1.8–60 pg/g. The usefulness of the developed method was tested by the analysis of real human samples, and several brominated flame retardants in different samples were detected and analyzed.     

Versatile lipid profiling by liquid chromatography–high resolution mass spectrometry using all ion fragmentation and polarity switching. Preliminary application for serum samples phenotyping related to canine mammary cancer

Lipids represent an extended class of substances characterized by such high variety and complexity that makes their unified analyses by liquid chromatography coupled to either high resolution or tandem mass spectrometry (LC–HRMS or LC–MS/MS) a real challenge. In the present study, a new versatile methodology associating ultra high performance liquid chromatography coupled to high resolution tandem mass spectrometry (UHPLC–HRMS/MS) have been developed for a comprehensive analysis of lipids. The use of polarity switching and “all ion fragmentation” (AIF) have been two action levels particularly exploited to finally permit the detection and identification of a multi-class and multi-analyte extended range of lipids in a single run. For identification purposes, both higher energy collision dissociation (HCD) and in-source CID (collision induced dissociation) fragmentation were evaluated in order to obtain information about the precursor and product ions in the same spectra. This approach provides both class-specific and lipid-specific fragments, enhancing lipid identification. Finally, the developed method was applied for differential phenotyping of serum samples collected from pet dogs developing spontaneous malignant mammary tumors and health controls. A biological signature associated with the presence of cancer was then successfully revealed from this lipidome analysis, which required to be further investigated and confirmed at larger scale.     

温州地区维持性血液透析患者矿物质代谢紊乱情况调查

目的 了解温州地区维持性血液透析患者矿物质代谢紊乱情况及相关影响因素。方法 收集2013 年在温州地区长期维持性血液透析患者血钙、血磷、全段甲状旁腺激素（iPTH）的资料,并根据美国肾脏病协会指南（KDOQI）及中华医学会肾脏病学分会（CSN）制订的《慢性肾脏病矿物质和骨异常诊治指导》计算患者的血钙、血磷、iPTH 的达标率,并对相关影响因素进行分析。结果 共收集2 227 例2013 年在温州地区各家透析中心维持性血液透析患者的资料。按KDOQI 指南标准,患者血钙、血磷、iPTH 的达标率分别为44.90%、46.30%、27.64%,3 项均达标者占6.29%;按《慢性肾脏病矿物质和骨异常诊治指导》建议,患者血钙、血磷、iPTH 的达标率分别为57.66%、46.30%、61.14%,3 项均达标者占16.97%。此外,三级医院维持性血液透析患者的平均血磷水平为（1.67±0.51）mmol/L,血磷达标率49.10%,二级医院患者的平均血磷水平为（1.79±0.53）mmol/L,达标率为43.22%,三级医院维持性血液透析患者的血磷达标率情况优于二级医院。血白蛋白、含钙磷结合剂的使用及透析频率是影响血钙达标的独立相关因素。患者的性别、年龄、活性维生素D 的使用、透析龄及透析频率是影响血磷达标的独立相关因素。患者活性维生素D的使用是影响iPTH 达标的独立相关因素。结论 温州地区维持性血液透析患者矿物质代谢紊乱纠正情况与有关指南的要求仍然存在较大差距;患者性别、营养状态、透析频率、含钙磷结合剂的服用、活性维生素D 的使用等是影响维持性血液透析患者矿物质代谢紊乱纠正的主要相关因素。     

Direct injection of biological fluids: Application of HPLC-analysis with precolumn enrichment to the pharmacokinetic studies of lonazolac

Summary Lonazolac and its p-hydroxylated metabolite M1 are determined in serum by high performance liquid chromatography using direct injection of comparatively large volumes with precolumn enrichment. No sample workup is required prior to analysis. Reproducibility, calibration plots, recoveries and detection limits are presented, together with various applications of the method. Data from direct injection of serum are also compared to results from a conventional extraction work-up.Presented at the 14th International Symposium on Chromagraphy London, September, 1982     

Enzyme-immunoassay (EIA) of Hepatitis B surface antigen (HBsAg) in microtiter plates

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Enzyme-immunoassay (EIA) of Hepatitis B surface antigen (HB_sAg) in microtiter plates