scholarly journals Optimization and Application of Electrochemical Transducer for Detection of Specific Oligonucleotide Sequence for Mycobacterium tuberculosis

Biosensors ◽  
2018 ◽  
Vol 8 (3) ◽  
pp. 84 ◽  
Author(s):  
Ricardo Corrêa ◽  
Filipe da Cruz ◽  
Cátia Santos ◽  
Thiago Pimenta ◽  
Diego Franco ◽  
...  

In this study, the electropolymerization of 4-hydroxyphenylacetic acid (4-HPA) over graphite electrodes (GE) was optimized, aiming its application as a functionalized electrochemical platform for oligonucleotides immobilization. It was investigated for the number of potential cycles and the scan rate influence on the monomer electropolymerization by using cyclic voltammetry technique. It was observed that the polymeric film showed a redox response in the region of +0.53/+0.38 V and the increase in the number of cycles produces more electroactive platforms because of the better electrode coverage. On the other hand, the decrease of scan rate produces more electroactive platforms because of the occurrence of more organized coupling. Scanning electron microscopy (SEM) images showed that the number of potential cycles influences the coverage and morphology of the electrodeposited polymeric film. However, the images also showed that at different scan rates a more organized material was produced. The influence of these optimized polymerization parameters was evaluated both in the immobilization of specific oligonucleotides and in the detection of hybridization with complementary target. Poly(4-HPA)/GE platform has shown efficient and sensitive for oligonucleotides immobilization, as well as for a hybridization event with the complementary oligonucleotide in all investigated cases. The electrode was modified with 100 cycles at 75 mV/s presented the best responses in function of the amplitude at the monitored peak current values for the Methylene Blue and Ethidium Bromide intercalators. The construction of the genosensor to detect a specific oligonucleotide sequence for the Mycobacterium tuberculosis bacillus confirmed the results regarding the poly(4-HPA)/GE platform efficiency since it showed excellent sensitivity. The limit of detection and the limit of quantification was found to be 0.56 (±0.05) μM and 8.6 (±0.7) μM, respectively operating with very low solution volumes (15 µL of probe and 10 µL target). The biosensor development was possible with optimization of the probe adsorption parameters and target hybridization, which led to an improvement in the decrease of the Methylene Blue (MB) reduction signal from 14% to 34%. In addition, interference studies showed that the genosensor has satisfactory selectivity since the hybridization with a non-specific probe resulted in a signal decrease (46% lower) when compared to the specific target.

2019 ◽  
Vol 8 (1) ◽  
pp. 542-550
Author(s):  
Bamdad Riahi-Zanjani ◽  
Mahdi Balali-Mood ◽  
Zarrin Es’haghi ◽  
Ahmad Asoodeh ◽  
Adel Ghorani-Azam

Abstract The ability of extraction and preconcentration of small amounts of substances from biological samples is very important in medical toxicology. On the other hand, minimal use of organic solvents is an important issue to prevent environmental damage. In the present study, we developed a new solid phase microextraction fiber using plant extracts as sorbent for extraction and preconcentration of morphine in urine sample. For this purpose, raw carbon nanotubes (CNTs) were functionalized with tobacco extracts. Functionalization was confirmed by Fourier transform infrared (FTIR) and Raman spectroscopy in addition to scanning electron microscope (SEM) images. The functionalized CNTs were coated on polypropylene hollow fiber. The results of HPLC analysis showed that the produced fiber could preconcentrate a very low concentration of morphine (0.25 ng/ml) in small volume of urine samples. Limit of detection (LOD) and limit of quantification (LOQ) for the produced fiber were determined 0.25 ng/ml and 0.825 ng/ml, respectively, and recovery of the fiber was determined 89% at LOQ. The produced fiber provided a recyclable and solvent free method for extraction of a trace amount of morphine, which can be successfully used for up to 30 times with no significant loss in the extraction efficiency.


2020 ◽  
pp. 3141-3153
Author(s):  
Asmaa Ghanim Dawood ◽  
Lazeeza Sattar Omer

A selective and sensitive spectrophotometric extraction method was established and used to estimate antihypertensive drug, losartan potassium. The method is based on the formation of blue ion pair of the anionic drug, losartan, and the cationic dye, methylene blue, at adjusted pH 6.5 in aqueous solutions, followed by quantitative extraction to dichloromethane;.The observed maximum absorbance was at π 654.9 nm. With 4.53321 x 105 M-1 cm-1 molar absorptivity, Beer's law was obeyed within a concentration range of 0.03-1.5 μg / ml. The limit of detection and the limit of quantification were 0.01μg / ml and 0.03μg / ml, respectively. The method's precision was estimated by a relative standard deviation (0.725%-1.64%), and the accuracy was validated at a recovery range (98.48-100.3%). The proposed method was successfully used for estimating losartan in the pharmaceutical formulation and in the urine sample.


2017 ◽  
Vol 71 (2) ◽  
pp. 97-104
Author(s):  
Sofija Rancic ◽  
Snezana Nikolic-Mandic ◽  
Aleksandar Bojic ◽  
Snezana Djoric-Veljkovic ◽  
Aleksandra Zarubica ◽  
...  

A new, simple, rapid, sensitive and selective kinetic spectrophotometric method for Pd(II) traces determination was elaborated in this paper. It is based on the catalytic effect of palladium ions upon the oxidation of methylene blue B (MBB) by (NH4)2S2O8 (APS) in citric buffer (BUF) solution. At the working temperature of 25 ?C and the wavelength of 662.4 nm, the optimal conditions for palladium determination were found. RSD value was found to be 2.6 to 4.9% for the investigated concentration range of Pd(II). Limit of detection (LD) was found to be 2.0 ng cm?3 Pd(II), and limit of quantification (LQ) is 6.9 ng cm?3 Pd(II). Only the presence of Pb2+, Hg2+ and Sn2+ in the reaction mixture interferes the determination of palladium by this method, while Au3+ and Ag+ have the catalytic effect upon the reaction rate. By this method, it is possible to determine the Pd(II) traces in the concentration range of 3.3x10?8 to 1.0x10?6 g cm?3. The method was confirmed by determination of Pd(II) traces in alloy PtRhG as well as in the powder of Pt. The obtained results were compared to those obtained by ICP-OES method and good agreement of results was found.


Author(s):  
Mohammad Hamzah Hamzah ◽  
Rawa M M Taqi ◽  
Muna M. Hasan ◽  
Raid J. M. Al-Timimi

A simple and accurate spectrophotometric method for the determination of Trifluoperazine HCl in pure and dosage forms was developed. The method is based on the reaction between Trifluoperazine HCl and p-chloroaniline in the presence of cerium ion as oxidizing agent which lead to the formation of violate color product that absorbed at a maximum wavelength 570nm while the blank solution was pink. Under the optimum conditions a linear relationship between the intensity and concentration of TRF in the range 4-50μg/ml was obtained . The molar absorptivity 3.74×103 L.mol-1.cm-1 , Limit of detection (2.21μg/ml), while limit of quantification was 7.39μg/ml. The proposed analytical method was compared with standard method using t-test and F-test , the obtained results shows there is no significant differences between proposed method and standard method. Based on that the proposed method can be used as an alternative method for the determination of TRF in pure and dosage forms.


Author(s):  
Sidra Amin ◽  
Amber R. Solangi ◽  
Dilawar Hassan ◽  
Nadir Hussain ◽  
Jamil Ahmed ◽  
...  

Background: In recent years, the occurrence and fate of environmental pollutants has been recognized as one of the emerging issues in environmental chemistry. A survey documented about a wide variety of these pollutants, which are often detected in our environment and these are major cause of shortened life spans and the global warming. These pollutants include toxic metal, pesticides, fertilizers, drugs and dyes released into soil and major water bodies. The presence of these contaminants causes major disturbance in eco-system’s balance. To tackle these issues many technological improvements are made to detect minute contaminations. The latest issue being answered by the scientists is the use of green nano materials as sensors which are economical, instant and give much better results at low concentrations and can be used for the field measurements resulting in no dangerous by-product that could lead to more environmental contamination. Nano materials are known for their wide band gap, enhanced physical and optical properties with option of tuneablity as per need, by optimizing certain parameters. They are proved to be good choice for analytical/optical sensors with high sensitivity. Objective: This review holds information about multiple methods that use green nanomaterials for the analytical assessment of environmental pollutants. UV-Vis spectrophotometry and electrochemical analysis using green and reproducible nanomaterials are the major focus of this review article. To date, there are number of spectrophotometric and electro chemical methods available that have been used for the detection of environmental pollutants such as toxic metals, pesticides and dyes. Conclusion: The use of nanomaterials can drastically change the detection limits due to having large surface area, strong catalytic properties, and tunable possibility. With the use of nano materials, lower than the marked limit of detection and limit of quantification were seen when compared with previously reported work. The used nano-materials could be washed, dried, and reused, which makes the methods more proficient, cost effective and environmentally friendly.


2020 ◽  
Vol 16 ◽  
Author(s):  
Nadereh Rahbar ◽  
Fatemeh Ahmadi ◽  
Zahra Ramezani ◽  
Masoumeh Nourani

Background: Sample preparation is one of the most challenging phases in pharmaceutical analysis, especially in biological matrices, affecting the whole analytical methodology. Objective: In this study, a new Ca(II)/Cu(II)/alginate/CuO nanoparticles hydrogel fiber (CCACHF) was synthesized through a simple, green procedure and applied for fiber micro solid phase extraction (FMSPE) of diazepam (DIZ) and oxazepam (OXZ) as model drugs prior to high-performance liquid chromatography-UV detection (HPLC-UV). Methods: Composition and morphology of the prepared fiber were characterized and the effect of main parameters on the fiber fabrication and extraction efficiency have been studied and optimized. Results: In optimal conditions, calibration curves were linear ranging between 0.1–500 µg L−1 with regression coefficients of 0.9938 and 0.9968. Limit of detection (LOD) (S/N=3) and limit of quantification (LOQ) (S/N=10) of the technique for DIZ and OXZ were 0.03 to 0.1 µg L−1. Within-day and between-day relative standard deviations (RSDs) for DIZ and OXZ were 6.0–12.5% and 3.3–9.4%, respectively. Conclusion: The fabricated adsorbent has been substantially employed to extraction of selected benzo-diazepines (BZDs) from human serum real specimens and the obtained recoveries were also satisfactory (82.1-109.7%).


2020 ◽  
Vol 16 (3) ◽  
pp. 277-286
Author(s):  
Amal A. El-Masry ◽  
Mohammed E. A. Hammouda ◽  
Dalia R. El-Wasseef ◽  
Saadia M. El-Ashry

Background: The first highly sensitive, rapid and specific green microemulsion liquid chromatographic (MELC) method was established for the simultaneous estimation of fluticasone propionate (FLU) and azelastine HCl (AZL) in the presence of their pharmaceutical dosage form additives (phenylethyl alcohol (PEA) and benzalkonium chloride (BNZ)). Methods: The separation was performed on a C18 column using (o/w) microemulsion as a mobile phase which contains 0.2 M sodium dodecyl sulphate (SDS) as surfactant, 10% butanol as cosurfactant, 1% n-octanol as internal phase and 0.3% triethylamine (TEA) adjusted at pH 6 by 0.02 M phosphoric acid; with UV detection at 220 nm and programmed with flow rate of 1 mL/min. Results: The validation characteristics e.g. linearity, lower limit of quantification (LOQ), lower limit of detection (LOD), accuracy, precision, robustness and specificity were investigated. The proposed method showed linearity over the concentration range of (0.5-25 µg/mL) and (0.1-25 µg/mL) for FLU and AZL, respectively. Besides that, the method was adopted in a short chromatographic run with satisfactory resolution factors of (2.39, 3.78 and 6.74 between PEA/FLU, FLU/AZL and AZL/BNZ), respectively. The performed method was efficiently applied to pharmaceutical nasal spray with (mean recoveries ± SD) (99.80 ± 0.97) and (100.26 ± 0.96) for FLU and AZL, respectively. Conclusion: The suggested method was based on simultaneous determination of FLU and AZL in the presence of PEA and BNZ in pure form, laboratory synthetic mixture and its combined pharmaceutical dosage form using green MELC technique with UV detection. The proposed method appeared to be superior to the reported ones of being more sensitive and specific, as well as the separation was achieved with good performance in a relatively short analysis time (less than 7.5 min). Highly acceptable values of LOD and % RSD make this method superior to be used in quality control laboratories with of HPLC technique.


Materials ◽  
2021 ◽  
Vol 14 (4) ◽  
pp. 723
Author(s):  
Kgotla K. Masibi ◽  
Omolola E. Fayemi ◽  
Abolanle S. Adekunle ◽  
Amal M. Al-Mohaimeed ◽  
Asmaa M. Fahim ◽  
...  

This report narrates the successful application of a fabricated novel sensor for the trace detection of endosulfan (EDS). The sensor was made by modifying a glassy-carbon electrode (GCE) with polyaniline (PANI), chemically synthesized antimony oxide nanoparticles (AONPs), acid-functionalized, single-walled carbon nanotubes (fSWCNTs), and finally, the AONP-PANI-SWCNT nanocomposite. The electrochemical properties of the modified electrodes regarding endosulfan detection were investigated via cyclic voltammetry (CV) and square-wave voltammetry. The current response of the electrodes to EDS followed the trend GCE-AONP-PANI-SWCNT (−510 µA) > GCE-PANI (−59 µA) > GCE-AONPs (−11.4 µA) > GCE (−5.52 µA) > GCE-fSWCNTs (−0.168 µA). The obtained results indicated that the current response obtained at the AONP-PANI-SWCNT/GCE was higher with relatively low overpotential compared to those from the other electrodes investigated. This demonstrated the superiority of the AONP-PANI-SWCNT-modified GCE. The AONP-PANI-SWCNT/GCE demonstrated good electrocatalytic activities for the electrochemical reduction of EDS. The results obtained in this study are comparable with those in other reports. The sensitivity, limit of detection (LoD), and limit of quantification (LoQ) of AONP-PANI-SWCNT/GCE towards EDS was estimated to be 0.0623 µA/µM, 6.8 µM, and 20.6 µM, respectively. Selectivity, as well as the practical application of the fabricated sensor, were explored, and the results indicated that the EDS-reduction current was reduced by only 2.0% when interfering species were present, whilst average recoveries of EDS in real samples were above 97%.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Colin Wood ◽  
Jason Sahl ◽  
Sara Maltinsky ◽  
Briana Coyne ◽  
Benjamin Russakoff ◽  
...  

Abstract Background Molecular assays are important tools for pathogen detection but need to be periodically re-evaluated with the discovery of additional genetic diversity that may cause assays to exclude target taxa or include non-target taxa. A single well-developed assay can find broad application across research, clinical, and industrial settings. Pathogen prevalence within a population is estimated using such assays and accurate results are critical for formulating effective public health policies and guiding future research. A variety of assays for the detection of Staphylococcus aureus are currently available. The utility of commercial assays for research is limited, given proprietary signatures and lack of transparent validation. Results In silico testing of existing peer-reviewed assays show that most suffer from a lack of sensitivity and specificity. We found no assays that were specifically designed and validated for quantitative use. Here we present a qPCR assay, SaQuant, for the detection and quantification of S. aureus as might be collected on sampling swabs. Sensitivity and specificity of the assay was 95.6 and 99.9 %, respectively, with a limit of detection of between 3 and 5 genome equivalents and a limit of quantification of 8.27 genome equivalents. The presence of DNA from non-target species likely to be found in a swab sample, did not impact qualitative or quantitative abilities of the assay. Conclusions This assay has the potential to serve as a valuable tool for the accurate detection and quantification of S. aureus collected from human body sites in order to better understand the dynamics of prevalence and transmission in community settings.


2020 ◽  
Vol 18 (1) ◽  
pp. 962-973
Author(s):  
Saira Arif ◽  
Sadia Ata

AbstractA rapid and specific method was developed for simultaneous quantification of hydrocortisone 21 acetate (HCA), dexamethasone (DEX), and fluocinolone acetonide (FCA) in whitening cream formulations using reversed-phase high-performance liquid chromatography. The effect of the composition of the mobile phase, analysis temperature, and detection wavelength was investigated to optimize the separation of studied components. The analytes were finally well separated using ACE Excel 2, C18 AR column having 150 mm length, 3 mm internal diameter, and 2 µm particle size at 35°C using methanol with 1% formic acid and double-distilled deionized water in the ratio of 60:40 (v/v), respectively, as the mobile phase in isocratic mode. Ten microliters of sample were injected with a flow rate of 0.5 mL/min. The specificity, linearity, accuracy, precision, recovery, limit of detection (LOD), limit of quantification (LOQ), and robustness were determined to validate the method as per International Conference on Harmonization guidelines. All the analytes were simultaneously separated within 8 min, and observed retention times of HCA, DEX, and FCA were 4.5, 5.5, and 6.9 min, respectively. The proposed method showed good linearity with the correlation coefficient, R2 = 0.999 over the range of 1–150 µg/mL for all standards. The linear regression equations were y = 12.7x + 118.7 (r = 0.999) for HCA, y = 12.9x + 106.8 (r = 0.999) for DEX, and y = 12.9x + 96.8 (r = 0.999) for FCA. The LOD was 0.25, 0.20, and 0.08 µg/mL for HCA, FCA, and DEX and LOQ was 2.06, 1.83, and 1.55 µg/mL for HCA, FCA, and DEX, respectively. The recovery values of HCA, DEX, and FCA ranged from 100.7–101.3, 102.0–102.6, and 100.2–102.0%, respectively, and the relative standard deviation for precision (intra- and interday) was less than 2, which indicated repeatability and reproducibility. The novelty of the method was described by forced degradation experimentation of all analytes in the combined form under acidic, basic, oxidative, and thermal stress. The proposed method was found to be simple, rapid, and reliable for the simultaneous determination of HCA, DEX, and FCA in cosmetics.


Sign in / Sign up

Export Citation Format

Share Document