scholarly journals Development of an Immunoassay for the Detection of Copper Residues in Pork Tissues

Biosensors ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 235
Author(s):  
Junqiu Zhang ◽  
Liwei Xu ◽  
Hongtao Jiang ◽  
Chuanlai Xu ◽  
Wenjing Liu ◽  
...  
Keyword(s):  
Ethylenediaminetetraacetic Acid ◽  
Limit Of Detection ◽  
Copper Chloride ◽  
Enzyme Linked Immunosorbent Assay ◽  
Limit Of Quantification ◽  
Detection Range ◽  
Cu Content ◽  
Cu Accumulation ◽  
Significant Difference ◽  
Pork Samples

The presence of high concentrations of copper (Cu) residues in pork is highly concerning and therefore, this study was designed to develop a high-throughput immunoassay for the detection of such residues in edible pork tissues. The Cu content in the pork samples after digestion with HNO3 and H2O2 was measured using a monoclonal antibody (mAb) against a Cu (II)–ethylenediaminetetraacetic acid (EDTA) complex. The resulting solution was neutralized using NaOH at pH 7 and the free metal ions in the solution were chelated with EDTA for the immunoassay detection. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) method was developed for Cu ion analysis. The half maximal inhibitory concentration of the mAb against Cu (II)–EDTA was 5.36 ng/mL, the linear detection range varied between 1.30 and 27.0 ng/mL, the limit of detection (LOD) was 0.43 μg/kg, and the limit of quantification (LOQ) was 1.42 μg/kg. The performances of the immunoassay were evaluated using fortified pig serum, liver, and pork samples and had a recovery rate of 94.53–102.24%. Importantly, the proposed immunoassay was compared with inductively coupled plasma mass spectroscopy (ICP-MS) to measure its performance. The detection correlation coefficients of the three types of samples (serum, pork, and liver) were 0.967, 0.976, and 0.983, respectively. Thirty pork samples and six pig liver samples were collected from local markets and Cu was detected with the proposed ic-ELISA. The Cu content was found to be 37.31~85.36 μg/kg in pork samples and 1.04–1.9 mg/kg in liver samples. Furthermore, we detected the Cu content in pigs with feed supplemented with tribasic copper chloride (TBCC) and copper sulfate (CS) (60, 110, and 210 mg/kg in feed). There was no significant difference in Cu accumulation in pork tissues between the TBCC and CS groups, while a remarkable Cu accumulation was found for the CS group in liver at 210 mg/kg, representing more than a two-fold higher level than seen in the TBCC group. Therefore, the proposed immunoassay was found to be robust and sensitive for the detection of Cu, providing a cost effective and practical tool for its detection in food and other complicated samples.

scholarly journals Gold-nanourchin complexed silicon dioxide-probe on gap-fingered interdigitated electrode surface for Parkinson’s Disease determination by current–volt measurement

2021 ◽  
Vol 11 ◽  
pp. 184798042098735
Author(s):  
Xiaohong Li ◽  
Wei Shi ◽  
Wenyan Zhang ◽  
Weiyao Chen ◽  
Dan Cao ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Silicon Dioxide ◽  
Limit Of Detection ◽  
Detection System ◽  
Interdigitated Electrode ◽  
Limit Of Quantification ◽  
Neurofilament Light Chain ◽  
Neurofilament Light ◽  
Detection Range

Parkinson’s disease (PD) is a nervous disorder, affects physical movement, and leads to difficulty in balancing, walking, and coordination. A novel sensor is mandatory to determine PD and monitor the progress of the treatment. Neurofilament light chain (NfL) has been recognized as a good biomarker for PD and also helps to distinguish between PD and atypical PD syndromes. Immunosensor was generated by current–volt measurement on gap-fingered interdigitated electrode with silicon dioxide surface to determine NfL level. To enhance the detection, anti-NfL antibody was complexed with gold-nanourchin and immobilized on the sensing electrode. The current–volt response was gradually increased at the linear detection range from 100 fM to 1 nM. Limit of detection and sensitivity were 100 fM with the signal-to-noise ratio at n = 3 on a linear curve ( y = 0.081 x + 1.593; R 2 = 0.9983). Limit of quantification falls at 1 pM and high performance of the sensor was demonstrated by discriminating against other neurogenerative disease markers, in addition, it was reproducible even in serum-spiked samples. This method of detection system aids to measure the level of NfL and leads to determine the condition with PD.

Serological biomarkers of zearalenone exposure in beef heifers receiving anti-mycotoxin additive

2020 ◽  
pp. 1-10
Author(s):  
C. Tonini ◽  
M.S. Oliveira ◽  
E.B. Parmeggiani ◽  
D.A.F. Sturza ◽  
A.O. Mallmann ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Biological Fluids ◽  
Basal Diet ◽  
Beef Heifers ◽  
Efficacy Evaluation ◽  
Limit Of Quantification ◽  
In Vivo Measurement ◽  
Significant Difference ◽  
Serological Biomarkers

The inclusion of anti-mycotoxin additives (AMA) in the diet of production animals has been widely used to avoid mycotoxin exposure. In order to confirm the efficacy of such products in vivo, measurement of mycotoxins and/or their metabolites in biological fluids is preconized. This study aimed at determining the serological biomarkers of zearalenone (ZEN), α-zearalenol, β-zearalenol, α-zearalanol, β-zearalanol (β-ZAL) and zearalanone, to evaluate the efficacy of an AMA in beef heifers. The trial lasted 37 days: 11 days of adaptation, 21 days of actual experiment, and 5 days of regression. Twenty-four heifers were randomly assigned to receive one of the following treatments (n=6/group): (T1) basal diet (control); (T2) basal diet + 5 mg/kg of ZEN; (T3) basal diet + 5 mg/kg of ZEN + 2.5 kg/t of AMA; and (T4) basal diet + 5 mg/kg of ZEN + 5.0 kg/t of AMA. Blood sampling was performed on different days after the diet was given. The samples were centrifuged to obtain the blood serum, and then analysed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). β-ZAL was detected above the limit of quantification both in the unconjugated (>0.60 ng/ml) and conjugated (>0.90 ng/ml) forms. The remaining metabolites presented concentrations under the limit of detection. In the efficacy evaluation of the AMA, there was no significant difference (P>0.05) between the treatments with and without additive at the tested levels of inclusion. Thus, β-ZAL may be employed as a biomarker of ZEN exposure via diet to evaluate the efficacy of an AMA through serological parameters. The technique applied in this study proved to be an adequate alternative for in vivo confirmation of the efficacy of products in adsorbing the toxin.

scholarly journals Streptavidin-conjugated gold nanoclusters as ultrasensitive fluorescent sensors for early diagnosis of HIV infection

2018 ◽  
Vol 4 (11) ◽  
pp. eaar6280 ◽  
Author(s):  
Aditya Dileep Kurdekar ◽  
L. A. Avinash Chunduri ◽  
C. Sai Manohar ◽  
Mohan Kumar Haleyurgirisetty ◽  
Indira K. Hewlett ◽  
...  
Keyword(s):  
Hiv Infection ◽  
Limit Of Detection ◽  
Gold Nanoclusters ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
Metal Nanoclusters ◽  
Detection Range ◽  
In Silico Studies ◽  
P24 Antigen ◽  
Hiv 1

We have engineered streptavidin-labeled fluorescent gold nanoclusters to develop a gold nanocluster immunoassay (GNCIA) for the early and sensitive detection of HIV infection. We performed computational simulations on the mechanism of interaction between the nanoclusters and the streptavidin protein via in silico studies and showed that gold nanoclusters enhance the binding to the protein, by enhancing interaction between the Au atoms and the specific active site residues, compared to other metal nanoclusters. We also evaluated the role of glutathione conjugation in binding to gold nanoclusters with streptavidin. As proof of concept, GNCIA achieved a sensitivity limit of detection of HIV-1 p24 antigen in clinical specimens of 5 pg/ml, with a detection range up to1000 pg/ml in a linear dose-dependent manner. GNCIA demonstrated a threefold higher sensitivity and specificity compared to enzyme-linked immunosorbent assay for the detection of HIV p24 antigen. The specificity of the immunoassay was 100% when tested with plasma samples negative for HIV-1 p24 antigen and positive for viruses such as hepatitis B virus, hepatitis C virus, and dengue. GNCIA could be developed into a universal labeling technology using the relevant capture and detector antibodies for the specific detection of antigens of various pathogens in the future.

scholarly journals Greener Monolithic Solid Phase Extraction Biosorbent Based on Calcium Cross-Linked Starch Cryogel Composite Graphene Oxide Nanoparticles for Benzo(a)pyrene Analysis

Molecules ◽  
2021 ◽  
Vol 26 (20) ◽  
pp. 6163
Author(s):  
Aree Choodum ◽  
Nareumon Lamthornkit ◽  
Chanita Boonkanon ◽  
Tarawee Taweekarn ◽  
Kharittha Phatthanawiwat ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Solid Phase Extraction ◽  
Limit Of Detection ◽  
Solid Phase ◽  
High Sensitivity ◽  
Rice Flour ◽  
Oxide Nanoparticles ◽  
Phase Extraction ◽  
Limit Of Quantification ◽  
Significant Difference

Benzo(a)pyrene (BaP) has been recognized as a marker for the detection of carcinogenic polycyclic aromatic hydrocarbons. In this work, a novel monolithic solid-phase extraction (SPE) sorbent based on graphene oxide nanoparticles (GO) in starch-based cryogel composite (GO-Cry) was successfully prepared for BaP analysis. Rice flour and tapioca starch (gel precursors) were gelatinized in limewater (cross-linker) under alkaline conditions before addition of GO (filler) that can increase the ability to extract BaP up to 2.6-fold. BaP analysis had a linear range of 10 to 1000 µgL−1 with good linearity (R2 = 0.9971) and high sensitivity (4.1 ± 0.1 a.u./(µgL−1)). The limit of detection and limit of quantification were 4.21 ± 0.06 and 14.04 ± 0.19 µgL−1, respectively, with excellent precision (0.17 to 2.45%RSD). The accuracy in terms of recovery from spiked samples was in the range of 84 to 110% with no significant difference to a C18 cartridge. GO-Cry can be reproducibly prepared with 2.8%RSD from 4 lots and can be reused at least 10 times, which not only helps reduce the analysis costs (~0.41USD per analysis), but also reduces the resultant waste to the environment.

Statistical Evaluation of Conventional and Portable Instrumentations for Cr(VI) Analysis on Chemistry Laboratory Waste Water

2020 ◽  
Vol 840 ◽  
pp. 406-411
Author(s):  
Suherman Suherman ◽  
Ghilma Milawonso ◽  
Kinichi Morita ◽  
Hitoshi Mizuguchi ◽  
Yuji Oki
Keyword(s):  
Waste Water ◽  
Inductively Coupled Plasma ◽  
Statistical Evaluation ◽  
Limit Of Detection ◽  
Liquid Waste ◽  
Chemistry Laboratory ◽  
Quality Of Data ◽  
Limit Of Quantification ◽  
Validation Methods ◽  
Significant Difference

The development of portable instrumentation for heavy metals analysis was increased rapidly. However, the quality of data from portable methods has so far been questioned when compared to conventional instrumentation. In this research, a comparative study of conventional and portable instrumentations for Cr(VI) analysis on liquid waste water of Chemistry Laboratory at Universitas Gadjah Mada (UGM) was conducted. This research started with validation and statistical evaluation of instrumentation methods which are UV-Visible spectrophotometer, portable spectrophotometer (PiCOEXPLORER) and Inductively Coupled Plasma Atomic Emission Spectrometer (ICP-AES). The validation methods consist of determination of linearity, sensitivity, limit of detection and limit of quantification. The results showed that the validation methods of ICP-AES were better than PiCOEXPLORER and UV-Vis spectrophotometer. Based on t-test, it was obtained that the result of Cr(VI) analyses with the comparison of UV-Vis and PiCOEXPLORER, ICP-AES and PiCOEXPLORER, and UV-Vis and ICP-AES; there were no significant difference (tcount< ttable). The ANOVA test (F test) results showed that the Fcount value is less than Ftable so that the results of Cr(VI) analysis in the standard solution and liquid waste samples measured by three instrumentations. Thus, it was concluded that portable instrumentations was good and gives the same quality as conventional instrumentations (UV-Vis and ICP AES).

HPTLC Separation of a Hepatoprotective Combination in Pharmaceutical Formulation and Human Plasma

2020 ◽  
Vol 58 (5) ◽  
pp. 411-417
Author(s):  
Maimana A Magdy ◽  
Rehab M Abdelfatah
Keyword(s):  
Human Plasma ◽  
High Performance ◽  
Limit Of Detection ◽  
Pharmaceutical Formulation ◽  
Limit Of Quantification ◽  
Ich Guidelines ◽  
Spiked Plasma ◽  
Significant Difference ◽  
Developing System

Abstract A binary mixture of Silymarin (SR) and Vitamin E (VE) acetate, of an antioxidant and a hepatoprotective effect, has been analyzed using a sensitive, selective and economic high performance thin layer chromatographic (HPTLC) method in their pure forms, pharmaceutical formulation and spiked human plasma. SR and VE were separated on 60F254 silica gel plates using hexane:acetone:formic acid (7:3:0.15, v/v/v) as a developing system with UV detection at 215 nm. The method was evaluated for linearity, accuracy, precision, selectivity, limit of detection (LOD) and limit of quantification (LOQ). SR and VE were detected in the linear range of 0.2–2.5 and 0.2–4.5 μg/band, respectively. Method validation was done as per ICH guidelines and acceptable results of accuracy of 99.86 ± 1.190 and 100.22 ± 1.609 for SR and VE, respectively were obtained. The method has been successfully applied for determination of the studied drugs in their pharmaceutical formulation without any interference from excipients, and in spiked plasma samples. Results obtained by the developed HPTLC-densitometric method were statistically compared to those obtained by the reported HPLC methods and no significant difference was found between them.

scholarly journals Stability-Indicating Photochemical Method for the Assay of Thiamine by Spectrophotometry

2018 ◽  
Vol 2018 ◽  
pp. 1-7 ◽  
Author(s):  
Iqbal Ahmad ◽  
Syed Haider Abbas ◽  
Zubair Anwar ◽  
Muhammad Ali Sheraz ◽  
Sofia Ahmed ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
The United States ◽  
Limit Of Quantification ◽  
Chromatography Method ◽  
Stability Indicating ◽  
Low Concentrations ◽  
Photochemical Method ◽  
Significant Difference ◽  
Liquid Chromatography Method ◽  
States Pharmacopeia

A stability-indicating photochemical method has been developed for the assay of thiamine (TH) salts in aqueous solution and in fresh and aged vitamin preparations. It is based on the photooxidation of TH by UV irradiation to form thiochrome (TC) in alkaline solution. The TC : TH ratio under controlled conditions of light intensity, temperature, pH, exposure time, and irradiation distance is constant and can be used to determine the concentration of UV irradiated TH solutions. TC, on extraction with isobutanol from the photodegraded solution of TH, has been determined by the UV spectrophotometric method at 370 nm. It exhibits a high intensity of absorption in the UV region that can be used for the assay of even low concentrations of TH. Under optimum conditions, Beer’s law is obeyed in the concentration range of 0.20–2.00 mg/100 ml (R2 = 09998). The limit of detection (LOD) and limit of quantification (LOQ) are 0.0076 and 0.0231 mg/100 ml, respectively. The method has been validated and applied to aqueous solutions and vitamin preparations. The results have statistically been compared with the United States Pharmacopeia liquid chromatography method. It has been found that there is no significant difference between the two methods at 95% confidence level.

scholarly journals Methodological aspects concerning sampling and determination of total selenium and selenium species in geothermal waters

2020 ◽  
Vol 18 (1) ◽  
pp. 5-16 ◽  
Author(s):  
Piotr Rusiniak ◽  
Anna Ruszczyńska ◽  
Katarzyna Wątor ◽  
Ewa Bulska ◽  
Ewa Kmiecik
Keyword(s):  
Limit Of Detection ◽  
Selenium Concentration ◽  
Geothermal Water ◽  
Limit Of Quantification ◽  
Geothermal Waters ◽  
Selenium Species ◽  
Icp Ms ◽  
Significant Difference ◽  
Total Selenium

AbstractThe work presents the results of geothermal water sample analysis with respect to the determination of total selenium concentration. For this purpose, geothermal water (GT) samples were collected from three different intakes (GT-1, GT-2, GT-3) with similar temperatures of about 85°C. Tests were carried out to see if the methodology of sample preparation influenced total selenium concentration during analysis by inductively coupled plasma mass spectrometry (ICP-MS). Samples (raw and filtered) were preserved with nitric acid (HNO3) and mineralised in the laboratory. From the data obtained it was found that there is no significant difference between total selenium concentration in raw and filtered samples. Following mineralisation, the concentrations in the samples were below the limit of detection or limit of quantification. While different analytical steps or procedures are applied, the results of total selenium concentration can vary. Furthermore, high-performance liquid chromatography coupled with ICP-MS was used for the identification of selenium species. The results revealed that hexavalent selenium – Se(VI) – in the geothermal water was found only in samples collected from the GT-2 and GT-3 intakes.

scholarly journals Adaptation of ELISA Detection of Plasmodium Falciparum and Plasmodium Vivax Circumsporozoite Proteins in Mosquitoes to a Multiplex Bead-Based Immunoassay

2021 ◽  
Author(s):  
Alice Sutcliffe ◽  
Seth R. Irish ◽  
Eric Rogier ◽  
Micaela Finney ◽  
Sarah Zohdy ◽  
...  
Keyword(s):  
Lower Limit ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Limits Of Detection ◽  
Detection Range ◽  
Cutoff Value ◽  
Sporozoite Rate ◽  
The Impact ◽  
Lower Limits ◽  
Multiplex Bead

Abstract Background: Plasmodium spp. sporozoite rates in mosquitoes are used to better understand malaria transmission intensity, the relative importance of vector species and the impact of interventions. These rates are typically estimated using an enzyme-linked immunosorbent assay (ELISA) utilizing antibodies against the circumsporozoite protein of P. falciparum (Pf), P. vivax VK210 (Pv210) or P. vivax VK247 (Pv247), employing assays that were developed over three decades ago. The ELISA method requires a separate assay plate for each analyte tested and can be time consuming as well as requiring sample volumes not always available. The bead-based multiplex platform allows simultaneous measurement of multiple analytes and may improve the lower limit of detection for sporozoites.Methods: Recombinant positive controls for Pf, Pv210 and Pv247 and previously developed circumsporozoite (cs) ELISA antibodies were used to optimize conditions for the circumsporozoite multiplex bead assay (csMBA) and to determine the detection range of the csMBA. After optimizing assay conditions, known amounts of sporozoites were used to determine the lower limit of detection for the csELISA and csMBA and alternate cutoff measures were applied to demonstrate how cutoff criteria can impact lower limits of detection. Sporozoite rates from 1275 mosquitoes collected in Madagascar and 255 mosquitoes collected in Guinea were estimated and compared using the established csELISA and newly optimized csMBA. All mosquitoes were tested (initial test), and those that were positive were retested (retest). When sufficient sample volume remained, an aliquot of homogenate was boiled and retested (boiled retest), to denature any heat-unstable cross-reactive proteins. Results: Following optimization of the csMBA, the lower limit of detection was 25 sporozoites per mosquito equivalent for Pf, Pv and Pv247 whereas the lower limits of detection for csELISA were found to be 1400 sporozoites for Pf, 425 for Pv210 and 1650 for Pv247. Combined sporozoite rates after re-testing of samples that initially tested positive for Madagascar mosquitoes by csELISA and csMBA were 1.4% and 10.3%, respectively, and for Guinea mosquitoes 2% by both assays. Boiling of samples followed by csMBA resulted in a decreased in the Madagascar sporozoite rate to 2.8-4.4% while the Guinea csMBA sporozoite rate remained at 2.0%. Using an alternative csMBA cutoff value of median fluorescence intensity (MFI) of 100 yielded a sporozoite rate after confirmational testing of 3.7% for Madagascar samples and 2.0% for Guinea samples. Whether using csMBA or csELISA, the following steps may help minimize false positives: specimens are appropriately stored and bisected anterior to the thorax-abdomen junction, aliquots of homogenate are boiled and retested following initial testing, and an appropriate cutoff value is determined.Conclusions: The csMBA is a cost-comparable and time saving alternative to the csELISA and may help eliminate false negatives due to a lower limit of detection, thus increasing sensitivity over the csELISA. The csMBA expands the potential analyses that can be done with a small volume of sample by allowing multiplex testing where analytes in addition to Pf, Pv210 and Pv247 can be added following optimization.

scholarly journals Evaluation of Improved and Current Vanillin Based Colorimetric Quantification Methods of Triterpenoids

2021 ◽  
Vol 10 (2) ◽  
Author(s):  
Rohit Suresh ◽  
Yash Kamdar ◽  
Vivek Garg ◽  
Angelina Kalathoti ◽  
Gayathri Renganathan
Keyword(s):  
Limit Of Detection ◽  
High Accuracy ◽  
Side Reactions ◽  
Improved Method ◽  
Limit Of Quantification ◽  
Detection Range ◽  
Quantification Method ◽  
Accuracy And Precision ◽  
Analyte Solution ◽  
Acidic Conditions

The most common colorimetric quantification method for triterpenoids utilizes vanillin in strongly acidic conditions to form a colored adduct with the analyte, and it has gained popularity because it is fast and easy to perform. Nevertheless, the detection range of this assay is limited and it is susceptible to side reactions, which cause an interference in the reading. Here, we quantify the rate of this interference in common solvents, and we present an alternative method to minimize the interference, which first incubates the analyte solution in perchloric acid at 60°C before the vanillin is added and finally quenched with acetic acid. We also successfully isolated the adduct that formed and confirmed that the color was due to vanillin polymerizing onto the analyte. The improved method had a correlation coefficient of 0.9959, as well as high accuracy and precision, which had a standard deviation of 0.072 mg/mL, and was not affected by minor changes in the conditions making it a robust method. Furthermore, our method had a limit of quantification of 2.37 mg/mL, which can analyze formulations with ease as well as a limit of detection of 0.782 mg/mL. Triterpenoids are quickly emerging as bioactive compounds, and as a result, our work improves upon current methods to quantify triterpenoids. 

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