Design an Enzyme-Less Sensitive Sensor for Cholesterol Detection Based on Poly Brucine/Polyvinylpyrrolidone Film

2020 ◽  
Vol 18 (11) ◽  
pp. 785-792
Author(s):  
Nahid Taghiee Karaji ◽  
Samineh Kaki ◽  
Arash Babakhanian

In the current study, the main focus is designing a new enzyme-less modified electrode based on the electro deposition of Poly Brucine/Polyvinylpyrrolidone on pencil graphite electrode and its utilizing for cholesterol quantification. The electrochemical responses of the proposed sensor were recorded using cyclic and square wave voltammetry techniques for detection of cholesterol at optimized pH =2. The morphology investigations of the modified surfaces were performed by scanning electron microscopy and the obtained images verified Nano formations in the structure of the sensor. The proposed electrochemical sensor revealed a good electro catalytic response to cholesterol concerning the parameters α = 0.83, log Ks = 3.11 and Γ = 8.55 × 10T8. Besides, the calibration plot was acquired with a linear portion in the concentration ranges of 25 to 95 nmol L−1 (nM) and the detection limit (LOD = 8.28 nM). The designed sensor demonstrated the reproducible and repeat-able outputs, the high sensitivity and stability in analyzing cholesterol as well as recoveries in blood serum samples without any side serious interferences.

2020 ◽  
Vol 16 (3) ◽  
pp. 341-348
Author(s):  
Surinya Traipop ◽  
Suchada Chuanuwatanakul ◽  
Orawon Chailapakul ◽  
Eakkasit Punrat

Background: Recently, Derris scandens, a Thai herbal medicine with anti-inflammatory activity, is widely used as beverage and supplementary food. When the traditional medicine is a choice for health therapy, the simple and reliable equipment is required to control the suitable consuming amount of the active component. Objective: To develop the electrochemical sensor for genistein determination in Derris scandens with high sensitivity and rapid operation. Methods: An in-house screen-printed electrochemical sensor consisting of a three-electrode system was developed for genistein determination. A silver/silver chloride (Ag/AgCl) reference electrode, a carbon counter electrode and a carbon working electrode were prepared on a 0.3-mm-thick plastic substrate by the screen-printing technique using conductive ink. The dimensions of each sensor were 2.5×1.0 cm. Only 50 µL of sample solution was required on this device for the determination of genistein concentration by rapid response square wave voltammetry. Results: The oxidation peak of genistein appeared with good response in acidic media at a peak potential of 0.6 V. Moreover, the signal was enhanced by modifying the conductive carbon ink with cobalt( II) phthalocyanine. Under the optimized conditions, the linear range was found to be 2.5-150 µM and the detection limit was 1.5 µM. Moreover, the small volume extraction was successfully developed without any further pre-concentration. This proposed method was applied to determine genistein in Derris scandens with satisfying results. Conclusion: The proposed method is promising as an alternative method for genistein determination with facile and fast analysis.


2016 ◽  
Vol 129 ◽  
pp. 205-212 ◽  
Author(s):  
Adrian Marcelo Granero ◽  
Gastón Darío Pierini ◽  
Sebastián Noel Robledo ◽  
María Susana Di Nezio ◽  
Héctor Fernández ◽  
...  

2017 ◽  
Vol 95 (12) ◽  
pp. 1267-1272 ◽  
Author(s):  
Zhiyou Xiao ◽  
Anjiang Tang ◽  
Hongsheng Huang ◽  
Ze Wang

A simple and sensitive assay for Ag+ was developed with unmodified gold nanoparticles (AuNPs) by using dynamic light scattering techniques. Ag+ could induce the oligonucleotide (5′-ATC ACT ATA TCA TAT ACT CAT-3′) to change from a single-stranded structure to a double-stranded structure and desorb from the surface of AuNPs, which triggered the aggregation of AuNPs in the salt solution. The average hydrodynamic diameter of aggregated AuNPs could be detected by using dynamic light scattering techniques. Under the optimum conditions, the average hydrodynamic diameter of AuNPs is proportional to the concentration of Ag+ within the range of 13.3–100.0 nmol/L, with a detection limit of 3.2 nmol/L. The method is easy to operate and has low sample consumption, high sensitivity and selectivity.


2018 ◽  
Vol 19 (11) ◽  
pp. 3485 ◽  
Author(s):  
Yunyun Luo ◽  
Yi He ◽  
Ditte Reker ◽  
Natasja Gudmann ◽  
Kim Henriksen ◽  
...  

N-terminal propeptide of type II collagen (PIINP) is a biomarker reflecting cartilage formation. PIINP exists in two main splice variants termed as type IIA and type IIB collagen NH2-propeptide (PIIANP, PIIBNP). PIIANP has been widely recognized as a cartilage formation biomarker. However, the utility of PIIBNP as a marker in preclinical and clinical settings has not been fully investigated yet. In this study, we aimed to characterize an antibody targeting human PIIBNP and to develop an immunoassay assessing type II collagen synthesis in human blood samples. A high sensitivity electrochemiluminescence immunoassay, hsPRO-C2, was developed using a well-characterized antibody against human PIIBNP. Human cartilage explants from replaced osteoarthritis knees were cultured for ten weeks in the presence of growth factors, insulin-like growth factor 1 (IGF-1) or recombinant human fibroblast growth factor 18 (rhFGF-18). The culture medium was changed every seven days, and levels of PIIBNP, PIIANP, and matrix metalloproteinase 9-mediated degradation of type II collagen (C2M) were analyzed herein. Serum samples from a cross-sectional knee osteoarthritis cohort, as well as pediatric and rheumatoid arthritis samples, were assayed for PIIBNP and PIIANP. Western blot showed that the antibody recognized PIIBNP either as a free fragment or attached to the main molecule. Immunohistochemistry demonstrated that PIIBNP was predominately located in the extracellular matrix of the superficial and deep zones and chondrocytes in both normal and osteoarthritic articular cartilage. In addition, the hsPRO-C2 immunoassay exhibits acceptable technical performances. In the human cartilage explants model, levels of PIIBNP, but not PIIANP and C2M, were increased (2 to 7-fold) time-dependently in response to IGF-1. Moreover, there was no significant correlation between PIIBNP and PIIANP levels when measured in knee osteoarthritis, rheumatoid arthritis, and pediatric serum samples. Serum PIIBNP was significantly higher in controls (KL0/1) compared to OA groups (KL2/3/4, p = 0.012). The hsPRO-C2 assay shows completely different biological and clinical patterns than PIIANP ELISA, suggesting that it may be a promising biomarker of cartilage formation.


2007 ◽  
Vol 2 ◽  
pp. 117727190700200 ◽  
Author(s):  
Ziad J. Sahab ◽  
Suzan M. Semaan ◽  
Qing-Xiang Amy Sang

Biomarkers are biomolecules that serve as indicators of biological and pathological processes, or physiological and pharmacological responses to a drug treatment. Because of the high abundance of albumin and heterogeneity of plasma lipoproteins and glycoproteins, biomarkers are difficult to identify in human serum. Due to the clinical significance the identification of disease biomarkers in serum holds great promise for personalized medicine, especially for disease diagnosis and prognosis. This review summarizes some common and emerging proteomics techniques utilized in the separation of serum samples and identification of disease signatures. The practical application of each protein separation or identification technique is analyzed using specific examples. Biomarkers of cancers of prostate, breast, ovary, and lung in human serum have been reviewed, as well as those of heart disease, arthritis, asthma, and cystic fibrosis. Despite the advancement of technology few biomarkers have been approved by the Food and Drug Administration for disease diagnosis and prognosis due to the complexity of structure and function of protein biomarkers and lack of high sensitivity, specificity, and reproducibility for those putative biomarkers. The combination of different types of technologies and statistical analysis may provide more effective methods to identify and validate new disease biomarkers in blood.


2021 ◽  
Vol 5 ◽  
pp. 239920262110550
Author(s):  
Joško Osredkar ◽  
Katja Krivic ◽  
Teja Fabjan ◽  
Kristina Kumer ◽  
Jure Tršan ◽  
...  

Aim: Although the levels of cardiac troponin I (cTnI) have proved to be a useful diagnostic biomarker of acute myocardial infarction, there are a wide variety of point-of-care (POC) analysers, which provide measurements of cTnI. The aim of this study was to compare the results obtained by the ADVIA Centaur ultra-assay cTnI assay (us-cTnI), ADVIA Centaur high-sensitive cTnI assay (hs-cTnI) and a POC high-sensitivity assay using PATHFAST. We also aimed to explore total turnaround time (TAT) for laboratory results using the POC PATHFAST analyser. Methods: Samples from 161 patients were taken. Of these samples, 129 were tested with all three assays (us-cTnI, hs-cTnI and PATHFAST), and 32 samples were tested on PATHFAST for the comparison of whole blood, serum and plasma. Results: Comparison of the POC testing methods in this study demonstrated that there are strong linear relationships between all three cTnI assays (us-cTnI, hs-cTnI and POC on PATHFAST). Furthermore, we also show there are strong linear relationships between the two high-sensitive cTnI assays (hs-cTnI and PATHFAST) for blood serum samples, as determined by Passing–Bablok regression analyses. In our comparison of our new data with our older study, the TAT went down. Conclusion: The timeliness of laboratory results is, in addition to accuracy and precision, one of the key indicators of laboratory performance, and at the same time has a significant impact on the course of the patient’s condition. It is therefore important that the laboratory strives to meet the expectations of clinicians regarding the time from the order to the result of the analysis.


2020 ◽  
Author(s):  
Beatriz Araujo Oliveira ◽  
Lea Campos de Oliveira ◽  
Franciane Mendes de Oliveira ◽  
Geovana Maria Pereira ◽  
Regina Maia de Souza ◽  
...  

AbstractBackgroundCOVID-19 disease (Coronavirus disease 2019) caused by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) is widespread worldwide, affecting more than 11 million people globally (July 6th, 2020). Diagnostic techniques have been studied in order to contain the pandemic. Immunochromatographic (IC) assays are feasible and low cost alternative for monitoring the spread of COVID-19 in the population.MethodsHere we evaluate the sensitivity and specificity of eleven different immunochromatographic tests in 98 serum samples from confirmed cases of COVID-19 through RT-PCR and 100 negative serum samples from blood donors collected in February 2019. Considering the endemic situation of Dengue in Brazil, we also evaluated the cross-reactivity with Dengue using 20 serum samples from patients with confirmed diagnosis for Dengue collected in early 2019 through four different tests.ResultsOur results demonstrated agreement between immunochromatographic assays and RT-PCR, especially after 10 days since the onset of symptoms. The evaluation of IgG and IgM antibodies combined demonstrated a strong level of agreement (0.85) of IC assays and RT-PCR. It was observed cross-reactivity between Dengue and COVID-19 using four different IC assays for COVID-19 diagnosis. The specificity of IC assays to detected COVID-19 IgM antibodies using Dengue serum samples varied from 80% to 85%; the specificity of IgG detection was 100% and total antibody was 95%.ConclusionsWe found high sensitivity, specificity and good agreement of IC assays, especially after 10 days onset of symptoms. However, we detected cross-reactivity between Dengue and COVID-19 mainly with IgM antibodies demonstrating the need for better studies about diagnostic techniques for these diseases.HighlightsImmunochromatographic assays demonstrated high sensitivity and specificity and good agreement with the gold-standard RT-PCR;Increase in sensitivity and specificity of assays using samples collected after the 10th day of symptoms;Cross-reaction with Dengue serology in evaluation of IgM.


Author(s):  
Rama Rao Nadendla ◽  
Patchala Abhinandana

Aim: The main aim of the current study is to give best and simple method for the estimation of antiparkinsonian drugs named Carbidopa, levodopa and entacapone. Study Design:  Simultaneous estimation of Carbidopa, levodopa and entacapone was performed by using Quadrapumped (SHIMADZU Prominance-i, LC-2030C) RP-HPLC equipped with PDA detector. Place and Duration of Study: Chalapathi Drug Testing Laboratory, Chalapathi Institute Of Pharmaceutical Sciences, Lam, Guntur-522034, Andhra Pradesh, India during the period of August 2019 to February 2020. Methodology: The assets of the study can determined as the process of qualification and quantification was done on SHIMADZU Prominance-i, LC-2030C system equipped with Phenomenex ODS (150 x 4.6 mm, 5μm) column and mobile phase was optimized using combination of acetonitrile and 0.1% ortho phosphoric acid in the ration of 50:50 v/v at a flow rate 1.0 ml/min. The wavelength was set as 270nm at ambient temperature by injecting 20µl of solution and the run time was fixed for 5 min. Results: Calibration plot shown best regression over the concentration range of 5-160 µg/ml of Carbidopa, Levodopa and Entacapone standard solutions. The LOD and LOQ were found to be 0.85 and 2.54 µg/ml for Entacapone, 0.24 and 0.71 µg/ml for Levodopa, 0.14 and 0.43 µg/ml for Carbidopa respectively. The accuracy of the proposed method was determined by performing recovery studies and was found to be between 98-102%. The repeatability testing for both sample and standard solutions was found as %RSD<2.0% which is within the acceptable limits showing that the method is precise as well. The proposed method was successfully applied for the marketed formulations of Carbidopa, Levodopa and Entacapone tablets. In addition the main feature of proposed method is economic and eco-friendly with less retention time around 5.0 min. Conclusion: Including all the optimized method parameters and statistical results given it can be concluded as a new, simple, sensitive, precise and accurate economical analytical method was developed and validated by RP-HPLC for the detection and quantification of Carbidopa, Levodopa and Entacapone which can be applied to the marketed formulation where there are no official compendial methods reported for this particular combination. The high sensitivity (LOD), mobile phase utilized and run time (=5) can be determined as an important features for this proposal.


PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9507
Author(s):  
Dandan Li ◽  
Jie Wu ◽  
Zhongjuan Liu ◽  
Ling Qiu ◽  
Yimin Zhang

Background Distinguishing between different types of thyroid cancers (TC) remains challenging in clinical laboratories. As different tumor types require different clinical interventions, it is necessary to establish new methods for accurate diagnosis of TC. Methods Proteomic analysis of the human serum was performed through data-independent acquisition mass spectrometry for 29 patients with TC (stages I–IV): 13 cases of papillary TC (PTC), 10 cases of medullary TC (MTC), and six cases follicular TC (FTC). In addition, 15 patients with benign thyroid nodules (TNs) and 10 healthy controls (HCs) were included in this study. Subsequently, 17 differentially expressed proteins were identified in 291 patients with TC, including 247 with PTC, 38 with MTC, and six with FTC, and 69 patients with benign TNs and 176 with HC, using enzyme-linked immunosorbent assays. Results In total, 517 proteins were detected in the serum samples using an Orbitrap Q-Exactive-plus mass spectrometer. The amyloid beta A4 protein, apolipoprotein A-IV, gelsolin, contactin-1, gamma-glutamyl hydrolase, and complement factor H-related protein 1 (CFHR1) were selected for further analysis. The median serum CFHR1 levels were significantly higher in the MTC and FTC groups than in the PTC and control groups (P < 0.001). CFHR1 exhibited higher diagnostic performance in distinguishing patients with MTC from those with PTC (P < 0.001), with a sensitivity of 100.0%, specificity of 85.08%, area under the curve of 0.93, and detection cut-off of 0.92 ng/mL. Conclusion CFHR1 may serve as a novel biomarker to distinguish PTC from MTC with high sensitivity and specificity.


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