scholarly journals Fluorescence-Based On-Resin Detection of Three Model Proteases

Chemosensors ◽  
2021 ◽  
Vol 9 (12) ◽  
pp. 359
Author(s):  
David Milićević ◽  
Jan Hlaváč
Keyword(s):  
Fluorescent Dyes ◽  
Simultaneous Detection ◽  
Solid Support ◽  
Buffer Solution ◽  
Solution Ph ◽  
New Approach ◽  
Backbone Amide ◽  
Cleavable Peptide ◽  
Backbone Amide Linker ◽  
Competitive Activities

A new approach to on-resin detection of three model proteases (trypsin, chymotrypsin, and thrombin) has been developed, while at the same time already described methodology for simultaneous detection of two enzymes (trypsin and chymotrypsin) has been additionally generalized. Appropriate immobilized substrates, comprising specifically cleavable peptide sequences capped with fluorescent dyes, have been synthesized on Rink Amide PEGA resin or Amino PEGA resin modified with backbone amide linker (BAL). Resulting solid support-bound probes were then dispersed into Tris-HCl buffer solution (pH = 8.0) and subjected to enzymatic cleavage. Liberated fluorophores have been tracked by fluorescence measuring. The competitive activities of studied proteases towards the thrombin probe have been efficiently limited and controlled by employing a Bowman-Birk inhibitor into a system.

scholarly journals Response Surface Optimization of an Extraction Method for the Simultaneous Detection of Sulfamethoxazole and 17β-Estradiol in Soil

Molecules ◽  
2020 ◽  
Vol 25 (6) ◽  
pp. 1415
Author(s):  
Rui Song ◽  
Qincheng Chen ◽  
Lili Yan ◽  
Pinhua Rao ◽  
Peng Sun ◽  
...  
Keyword(s):  
Response Surface ◽  
High Performance ◽  
Detection Method ◽  
Simultaneous Detection ◽  
Animal Husbandry ◽  
Ultrasound Assisted Extraction ◽  
Buffer Solution ◽  
Solution Ph ◽  
Simultaneous Extraction ◽  
17Β Estradiol

Antibiotics and hormones widely exist in fertilizers and manures, which are excessively used in agriculture and animal husbandry. Considering their potential harm to the environment and human health, the detection of antibiotics and hormones has become a necessity. However, current methods find it difficult to simultaneously extract and detect antibiotics and hormones in soil and to maintain a high level of accuracy and a low cost. In this study, a straightforward, convenient, and simultaneous extraction and detection method of a representative antibiotic (sulfamethoxazole, SMZ) and hormone (17β-Estradiol, E2) in soil has been established. Ultrasound-assisted extraction (UAE) was used in the pretreatment process and high-performance liquid chromatography with the ultraviolet detector (HPLC-UV) method was then chosen in the detection process. By means of single factors and response surface experiments, optimal extraction conditions were a 41-mL buffer solution (pH 4.27) mixed with 1 g of soil sample, an ultrasonication time of 36 min, an ultrasonication temperature of 25 °C, and two extraction cycles. The detection limits of 0.3–10 μg/kg and quantification limits of 1–30 μg/kg have been obtained. Finally, the optimized simultaneous extraction and detection method was validated by three different real soil samples with recoveries ranging from 79.49% to 86.47%.

Determination of Synephrine and Naringenin in Aifunuangong Pills by High Performance Capillary Electrophoresis

2013 ◽  
Vol 850-851 ◽  
pp. 1160-1163
Author(s):  
Hai Xing Liu ◽  
Qing Liu ◽  
Yi Min Feng ◽  
Lin Tong Wang ◽  
Yan Fu Huan ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
High Performance ◽  
Buffer Solution ◽  
Solution Ph ◽  
New Approach ◽  
Capillary Electrophoresis Method ◽  
Electrophoresis Method ◽  
High Performance Capillary Electrophoresis ◽  
Separation Of Components

This paper established high performance capillary electrophoresis method to determine synephrine and naringenin in aifunuangong pills. In 30mmol/L Na2B4O7 buffer solution (pH=10.00), effective separation of components to be tested was achieved, which provided a new approach of quality control of Chinese medicine aifunuangong pills.

scholarly journals Simultaneous Adsorptive Stripping Voltammetric Analysis of Heavy Metals at Graphenated Cupferron Pencil Rods

2022 ◽  
Author(s):  
Nelia Abraham Sanga ◽  
Nazeem Jahed ◽  
Zandile Leve ◽  
Emmanuel Iwuoha ◽  
Keagan Pokpas
Keyword(s):  
Metal Ion ◽  
Tap Water ◽  
Low Cost ◽  
Simultaneous Detection ◽  
Chelating Agent ◽  
Acetate Buffer Solution ◽  
Buffer Solution ◽  
Solution Ph ◽  
Voltammetric Analysis ◽  
Electrochemically Reduced Graphene Oxide

Abstract Electroanalysis of heavy metal ions in the presence of cupferron ligands has been extensively studied due to its ability to form stable metallic coordination complexes. Herein, electrochemically reduced graphene oxide (ERGO) sheets were for the first time employed in conjunction with low-cost, disposable pencil graphite rods and in-situ plated thin mercury films (HgF) for the simultaneous detection of Cd2+, Cu2+, Pb2+, and Zn2+ in the presence of cupferron as a chelating agent by square-wave adsorptive cathodic stripping voltammetry (SW-AdCSV). The technique is based on the catalytic reduction of adsorbed cupferron-metal ion complexes at the surface of the ERGO-HgF-PGE at 0.1 V for 60 s in 0.1 M acetate buffer solution (pH 4.6). Owing to the improved electronic and surface effects associated with ERGO inclusion, improved sensitivity was further achieved. Under optimized conditions, the ERGO-HgF-PGE showed a linear relationship from 20 to 200 µg.L-1 with detection limits below the US-EPA of 0.17 μg.L-1 , 0.02 μg.L-1 , 0.17 μg.L-1 and 0.14 μg.L-1 for Cd2+, Cu2+, Pb2+ and Zn2+, respectively at a deposition time of 60 s. The ERGO-HgF-PGE exhibited highly reproducible results with negligible intermetallic interferences and applied successfully to the determination of trace metals in tap water with satisfactory results.

Detection of Hepatitis a Virus in Drinking Water by Enzyme -Immunoassay Using Ultracentrifugation for Virus Concentration

1985 ◽  
Vol 17 (10) ◽  
pp. 39-41 ◽  
Author(s):  
A. Schnattinger
Keyword(s):  
Membrane Filtration ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Phosphate Buffer Solution ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Concentration ◽  
Buffer Solution ◽  
Solution Ph ◽  
Two Stage

Ten litres of tapwater were seeded with 200 µl (8×108 HAV particles) of a commercial (Organon Teknika) suspension of hepatitis A virus. Following WALTER and RÜDIGER (1981), the contaminated tapwater was treated with a two-stage technique for concentration of viruses from solutions with low virus titers. The two-stage technique consists of aluminium hydroxideflocculation (200 mg/l Al2(SO4)3. 18 H2O, pH 5,4-5,6) as first stage, the second stage of a lysis of aluminium hydroxidegel with citric acid/sodium citrate-buffer (pH 4,7; 1 ml/l sample), separation of viruses from the lysate by ultracentrifugation and suspension in 1 ml phosphate buffer solution (pH 7,2). A commercial solid phase enzyme-linked immunosorbent assay (ELISA) was used for the detection of HAV. HAV was detecterl in the 10.000:1 concentrates, but not in the seeded 101 samples. Approximately 4×108 of the inoculated 8×108 HAV particles were found in the 1 ml concentrates. The efficiency of detection is about 50%, the virus concentration 5000-fold. Although the percentage loss of HAV in comparison with concentration by means of membrane filtration is similar, the ultracentrifugation method yields a larger sample/concentrate ratio, so that smaller amounts of HAV can be detected more efficiently because of the smaller end-volume.

Development and Validation of Stability Indicating HPLC Method for the Determination of Impurities in the Sterile Mixture of Cefoperazone and Sulbactam

2019 ◽  
Vol 15 (7) ◽  
pp. 762-775
Author(s):  
Ramu Ivaturi ◽  
Thuttagunta Manikya Sastry ◽  
Satyaveni Sunkara
Keyword(s):  
Quantitative Estimation ◽  
Ammonium Hydroxide ◽  
Hplc Method ◽  
Forced Degradation ◽  
Buffer Solution ◽  
Solution Ph ◽  
Pharmaceutical Dosage Forms ◽  
Stability Indicating ◽  
Validation Parameters ◽  
Abdominal Infections

Background: Cefoperazone Sulbactam injection is a cephalosporin antibiotic with a β- lactamase inhibitor used in the treatment for intra abdominal infections, Urinary track infections, surgical infections, etc. The combination is not official in any of the pharmacopeia for their content and impurities determination. Introduction: The present study involves the development of a simple, rapid, accurate, sensitive and stability indicating RP-HPLC method for the quantitative estimation of Cefoperazone Sulbactam mixture and its impurities in bulk and pharmaceutical dosage forms. Methods: 0.005 M Tetrabutyl ammonium hydroxide buffer solution pH adjusted to 6.80 and Acetonitrile combination has been used in a gradient programme with a flow rate of 1.0 ml/min. The retention time of Cefoperazone and Sulbactam were observed at around 8.5 and 19.5 minutes respectively. The UV detection was carried out at a wavelength of 230 nm. The chromatographic separation was achieved using Waters xbridge C18-150*4.6 mm, 3.5 µm HPLC column. The method has been validated according to the current International Council for Harmonization (ICH) guidelines for the method validation parameters such as Specificity, linearity, range, accuracy, precision, robustness and sensitivity. Results: The validation results indicate that the method is specific, as the known impurities and other impurities formed during the forced degradation studies were not co-eluting with the main components. Moreover, all these impurities were found to be spectrally pure, proving the stability indicating power of the method. The linearity and range of the method is in the range of 0.01-150%, highly accurate (100.2%), precise (<1%) and robust. Conclusion: The proposed method was accurate and specific for the quantitative analysis of Cefoperazone and Sulbactam and their related impurities in the sterile mixture. Hence the proposed method can be used for the quantification of impurities in routine as well as stability analysis in the development as well as quality control laboratories.

Non-enzymatic determination of purine nucleotides using a carbon dot modified glassy carbon electrode

2019 ◽  
Vol 11 (30) ◽  
pp. 3866-3873 ◽  
Author(s):  
R. Karthikeyan ◽  
D. James Nelson ◽  
S. Abraham John
Keyword(s):  
Glassy Carbon ◽  
Low Cost ◽  
Phosphate Buffer Solution ◽  
Purine Nucleotides ◽  
Buffer Solution ◽  
Solution Ph ◽  
Carbon Dot ◽  
Enzymatic Determination ◽  
Sensitive Determination

Selective and sensitive determination of one of the purine nucleotides, inosine (INO) using a low cost carbon dot (CD) modified glassy carbon (GC) electrode in 0.2 M phosphate buffer solution (pH 7.2) was demonstrated in this paper.

scholarly journals Electro-Polymerized Titan Yellow Modified Carbon Paste Electrode for the Analysis of Curcumin

Surfaces ◽  
2021 ◽  
Vol 4 (3) ◽  
pp. 191-204
Author(s):  
Edwin S. D’Souza ◽  
Jamballi G. Manjunatha ◽  
Chenthattil Raril ◽  
Girish Tigari ◽  
Huligerepura J. Arpitha ◽  
...  
Keyword(s):  
Carbon Paste Electrode ◽  
Limit Of Detection ◽  
Phosphate Buffer Solution ◽  
Modified Carbon Paste Electrode ◽  
Carbon Paste ◽  
Buffer Solution ◽  
Solution Ph ◽  
Real Sample Analysis ◽  
Titan Yellow ◽  
Paste Electrode

A modest, efficient, and sensitive chemically modified electrode was fabricated for sensing curcumin (CRC) through an electrochemically polymerized titan yellow (TY) modified carbon paste electrode (PTYMCPE) in phosphate buffer solution (pH 7.0). Cyclic voltammetry (CV) linear sweep voltammetry (LSV) and differential pulse voltammetry (DPV) approaches were used for CRC detection. PTYMCPE interaction with CRC suggests that the electrode exhibits admirable electrochemical response as compared to bare carbon paste electrode (BCPE). Under the optimized circumstances, a linear response of the electrode was observed for CRC in the concentration range 2 × 10−6 M to 10 × 10−6 M with a limit of detection (LOD) of 10.94 × 10−7 M. Moreover, the effort explains that the PTYMCPE electrode has a hopeful approach for the electrochemical resolution of biologically significant compounds. Additionally, the proposed electrode has demonstrated many advantages such as easy preparation, elevated sensitivity, stability, and enhanced catalytic activity, and can be successfully applied in real sample analysis.

scholarly journals Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections.

1993 ◽  
Vol 41 (11) ◽  
pp. 1599-1604 ◽  
Author(s):  
S R Shi ◽  
B Chaiwun ◽  
L Young ◽  
R J Cote ◽  
C R Taylor
Keyword(s):  
Androgen Receptor ◽  
Enzymatic Digestion ◽  
Antigen Retrieval ◽  
Urea Solution ◽  
Paraffin Sections ◽  
Buffer Solution ◽  
Solution Ph ◽  
Citrate Buffer ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed

We developed a staining protocol for demonstration of androgen receptor (AR) in formalin-fixed, paraffin-embedded tissue sections. The method is based on the antigen retrieval microwave (MW) heating technique. Results are compared with different types of enzyme digestion pre-treatments. The strongest immunostaining signal and clearest background were obtained by MW heating of dewaxed paraffin sections in 5% urea or citrate buffer solution (pH 6); pure distilled water gave less consistent results. Enzymatic digestion with pepsin (0.05% in 2 N HCl) for 30 min at room temperature, or trypsin followed by pronase, or pronase digestion alone, also produced enhanced staining of AR in some cases, but there was more nonspecific background, and specific reactivity was less intense. The antigen retrieval MW method can be used to demonstrate AR epitope in prostate tissue after fixation in formalin for as long as 7 days. AR immunolocalization was also compared in frozen and paraffin sections processed from the same specimen of prostate carcinoma tissue and was found to be qualitatively and quantitatively similar. This study also provided new information concerning the basic principles of the antigen retrieval MW method that may be helpful in further development of this technique.

Surface Modification of Nanosized Biphasic α-TCP/HA Powders

2007 ◽  
Vol 330-332 ◽  
pp. 223-226 ◽  
Author(s):  
Li Li Pan ◽  
Yan Bao Li ◽  
Chao Zou ◽  
Wen Jian Weng ◽  
Kui Cheng ◽  
...  
Keyword(s):  
Surface Modification ◽  
Stearic Acid ◽  
Tricalcium Phosphate ◽  
Calcium Phosphates ◽  
Biological Characteristics ◽  
Buffer Solution ◽  
Solution Ph

Stearic acid was utilized to modify biphasic alpha-tricalcium phosphate (α-TCP)/hydroxyapatite (HA) powders in the ethanol. The results showed that the dispersion of biphasic α-TCP/HA powders (BCPs) in non-polar matrix improved. And the released content of Ca2+ and PO4 3- of the BCPs soaked in the NaAc-HAc buffer solution (pH 5.0) was almost same as that before modification. Stearic acid could modify the suface properties of BCPs and would not obviously affect their biological characteristics, which affords a good groundwork of application of calcium phosphates powders.

Controlled Release of Active Substance from Biodegradable Copolymer Matrix

2006 ◽  
Vol 510-511 ◽  
pp. 798-801
Author(s):  
Hyung Suk So ◽  
Hyun Chul Shin ◽  
Beom Suk Kim ◽  
Yeong Seok Yoo
Keyword(s):  
Methylene Blue ◽  
Active Substance ◽  
Phosphate Buffer Solution ◽  
Ultra Violet ◽  
Hydroxyl Groups ◽  
Buffer Solution ◽  
Solution Ph ◽  
Effective Discharge ◽  
Constant Rate ◽  
Long Time

The purpose of this study is to develop a new system to control effective discharge of active substances such as agricultural chemicals. To synthesize a naturally dissolvable polymer; ε-caprolactone and diglycolide were copolymerized with ethylene glycol as an initiator to produce macrodiol. As macrodiol has hydroxyl groups in both ends, they are modified with methacryloyl chloride for photochemical networking. After standard macromonomer produced by this procedure was physically mixed with methylene blue, it was networked with ultra-violet rays to be filmed. This film is naturally dissolvable and hydrolytic. As a result of hydrolytic test with a crosslinked structure of 10 % methylene blue, it decreased by 9 % for seven weeks in 37 °C phosphate buffer solution (pH = 7). Thus, we verified that active substance can be discharged from a crosslinked structure for a long time at a constant rate under room temperature.

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