Method For Determination Of Tetracycline Research Articles

Evolution of a natural TetR protein and development of a Fe3O4 assisted semi-homogeneous fluorescent method for determination of tetracyclines in milk

Compared with antibody, the recognition spectrum of a receptor is broader, and its recognition ability can be improved using simple mutagenesis technique. Compared with conventional immunoassay, the magnetic bead based immunoassay is simpler and can be recycled. Compared with colorimetric and luminescent immunoassays, fluoroimmunoassay is simpler because it does not require a substrate. So a method combines these merits is desirable. In this study, two amino acids in the binding pocket of a natural Escherichia coli TetR protein were mutated to produce a mutant, and the molecular docking showed the binding energies and the numbers of contact acid for 10 tetracyclines all increased. The mutant was coupled with Fe3O4 to synthesize a magnetic complex, and a fluorescent tracer was synthesized by coupling quantum dot and minocycline with bovine serum albumin. Under the assistance of 96-well bottom magnet, a semi-homogeneous method based on the two materials was developed on conventional microplate for determination of the 10 tetracyclines in milk. Results showed once assay was finished within 20 min, the limits of detection (drug concentration showing 10% inhibition) for the 10 drugs were in the range of 0.32–0.94 ng/mL, and the magnetic complex could be regenerated for 6 times. Furthermore, the sensitivities were improved for 4–6 folds in comparison with the use of natural TetR. Therefore, this method is simple, sensitive, time-saving and recyclable, and it can be used for routine screening of the 10 tetracyclines in milk.

Evaluation of analytical method for determination of tetracyclines in their dosage forms by HPLC using different types of chromatographic stationary phases (RP-C8 and RP-C18)

Evaluation of analytical method for determination of tetracyclines in their dosage forms by HPLC using different types of chromatographic stationary phases (RP-C8 and RP-C18)

Adsorptive stripping voltammetric determination of Tetracycline in pharmaceutical capsule formulation using Poly(Malachite green) modified glassy carbon electrode

A selective and sensitive electrochemical method based on glassy carbon electrode modified with poly(malachite green) was developed for determination of tetracycline in pharmaceutical capsule formulation. Cyclic voltammetry and electrochemical impedance spectroscopy using [Fe(CN)6]3-/4- as a probe were used to characterize the potentiodynamiclly deposited poly(malachite green) on the surface of glassy carbon electrode. In contrast to the unmodified glassy carbon electrode, the fabricated poly(malachite green) modified glassy carbon electrode showed catalytic property towards two steps irreversible oxidation of tetracycline. Better correlation of the oxidative peak current with the scan rate than with the square root of scan rate supported by slope of 0.60 for log(current) versus log(scan rate) indicated that the oxidation reaction of tetracycline at the modified electrode was predominantly controlled by electron exchange step at the solution polymer interface. Under optimized solution pH, and accumulation parameters, the square wave adsorptive anodic striping peak current response of the modified electrode showed linear dependence on concentration of tetracycline in the range 5–100 μM with determination coefficient, method detection limit, and quantification limit of 0.99588, 1.6 μM, and 5.3 μM, respectively. The tetracycline content of a capsule sample claimed to have 250 mg/capsule was found to be 250.53 mg/capsule with 0.21% deviation. Excellent spike recovery result of 99.80%, and 98.49–99.78% recovery of tetracycline in capsule sample in the presence of 50–200% of UA, AA, and ampicillin validated the applicability of the method for determination of tetracycline in real samples with complex matrix like capsule formulations.

Spectrophotometric Determination of Tetracycline, in pharmaceutical formulations by oxidative coupling

This research includes the development of spectrophotometric method for determination of Tetracycline via oxidative coupling with the N,N-Diethyl-p-phenylene diamine reagent in presence of oxidizing agentN-Bromosuccinimide in the alkaline medium oxidation time 15 min to form a colored-dye violet, which dissolved in water and showed the highest intensity of absorption at λmax 552nm, the method has been obeyed Beer's Law in the concentration range (10-32μg. ml-1), with molar absorptivity of 10579 L. mol-1. Cm-1, Sandel Index value 0.0454 μg. cm-2, the detection limited 0.329 μg. ml-1, with correlation coefficient 0.9942 , the relative standard deviation of the method does not exceed 0.302% the proposed method was applied successfully for the determination of Tetracycline Hydrochloride on pharmaceutical preparation by both the direct and standard addition methods for three concentrations, the recovery ranged between 98.8-102.71%. the evaluations of results of this method was a achieved applying t and F tests. The results showed that there is no significant difference between the proposed and standard method, and thus the proposed method is of good validity for the determination Tetracycline in its pharmaceutical preparations

A simple kinetic spectrophotometric method for simultaneous determination of tetracyclines by use of chemometrics

A new kinetic spectrophotometric method for simultaneous determination of chlortetracycline, tetracycline and oxytetracycline in real samples has been researched and developed. The method was based on the kinetic reaction involving the oxidation of the drugs by alkaline potassium permanganate at 40 °C. After the addition of these drugs, an increasing absorbance at 526 nm and a decreasing absorbance at 608 nm caused by manganate and permanganate, respectively, were observed. The three-way kinetic-spectral data in the 325–725 nm range from 0 to 360 s were collected and applied to build chemometrics models. The plot of absorbance versus concentration was rectilinear over the 0.05–0.75 µg mL−1 range for chlortetracycline and tetracycline, and 0.02–0.34 µg mL−1 for oxytetracycline, with the limit of detection of 0.013, 0.015 and 0.011 µg mL−1, respectively. The experimental conditions were carefully investigated to optimize the development and stability of the color products (potassium manganate). The proposed method was successfully applied to the simultaneous determination of tetracyclines (TCs) in feed and muscle samples, and the results showed satisfactory agreement with those from of the HPLC method.

A HPLC with fluorescence detection method for the determination of tetracyclines residues and evaluation of their stability in honey

A high-performance liquid chromatography with fluorescence detection method for the simultaneous determination of oxytetracycline (OTC), tetracycline (TC) and chlortetracycline (CTC) residues in honey was developed and validated. Sample preparation was done in a single solid phase extraction step. The chromatographic separation was performed on a C 8 column. Matrix matched calibration curves showed linearity higher than 0.99. The accuracy values lay between 86% and 111% and the precision was lower than 11%. Limits of detection and quantitation were 8 μg kg −1 and 25 μg kg −1, respectively. The method was employed to evaluate the stability of the tetracyclines in honey during 60 days of storage. The residue levels of OTC, TC and CTC were reduced 97%, 76% and 71%, respectively. Honey samples ( n = 22) collected from the retail market were analyzed. No tetracyclines residues were detected.

Determination of tetracyclines in food samples by molecularly imprinted monolithic column coupling with high performance liquid chromatography

A novel solid phase extraction (SPE) method for determination of tetracyclines (TCs) in milk and honey samples by molecularly imprinted monolithic column was developed. Using tetracycline (TC) as the template, methacrylic acid (MAA) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the cross-linker, methanol as the solvent, cyclohexanol and dodecanol as the mixed porogenic solvents, a TC imprinted monolithic column was prepared by in situ molecular imprinting technique for the first time, and the optimal synthesis conditions and the selectivity of TC imprinted monolithic column were investigated. The interfering substances in food samples and TCs can be separated successfully on imprinted column. Molecularly imprinted solid phase extraction (MISPE) coupling with C18 column was used to determinate the TCs in milk and honey. The recoveries of this method for six tetracyclines antibiotics such as tetracycline (TC), oxytetracycline (OTC), minocycline (MINO), chlortetracycline (CTC), metacycline (MTC) and doxycycline (DTC) were investigated, and high recoveries of 73.3–90.6% from milk samples and 62.6–82.3% from honey samples were obtained. A method for determination of TCs at low concentration level in milk and honey samples was successfully developed by using the monolithic column as the precolumn for solid phase extraction of six TCs compounds.

Analysis of Certain Tetracyclines and Oxytetracyclines through Charge Transfer Complexation

Problem statement: Several studies were carried out to develop and validate a simple, rapid and accurate spectrophotometric method for the analysis of tetracycline (TC) and oxytetracycline (OTC) in the pure form and in pharmaceutical dosage forms, despite the varieties of these analytical techniques, the literature revealed that no spectrophotometric method for determination of tetracycline and oxytetracycline using charge transfer complexation with chloranilic acid has been reported. Approach: In the present study, a new and simple spectrophotometric method is described for the determinations of tetracycline (TC) and oxytetracycline (OTC), the method is based on the molecular interaction between the tetracyclines and Chloranilic Acid (CA), to form a charge-transfer complex in which the drugs acts as n-donor and chloranilic acid as I€-acceptor in acetonitrile. The spectra, various experimental parameters for the reaction, the stability of the complex and the validty of the method were investigated. Results: The formed complex was found to absorb maximally at 540nm, Beer’s law is obeyed in the concentration ranges 2.5-30 µg mL-1 and 5.0-40 µg mL-1 for TC and OTC, respectively. The limits of detection (S/N = 3) were 0.40 µg mL-1 for tetracycline and 0.50 µg mL-1 for oxytetracycline. The molar ratio was found to be approximately 1:1, confirming that one molecule of tetracycline or oxytetracycline combines with one molecule of chloranilic acid. Conclusions and Recommendations: The proposed method was found to be rapid, accurate and sensitive and may be applied for estimation of named drugs in pharmaceutical dosage forms without interferences from the common additives encountered. Percentage recoveries ranged from 99.17%±0.95-100.38%±0.82. From the statistical analysis of the obtained results one can conclude that no significant difference between the proposed method and other official methods as evident from the t-test and variance ratio. These findings recommend the utility of the proposed method in the quality control analysis of TC and OTC in the pure form and pharmaceutical dosage forms.

ローヤルゼリー中のテトラサイクリン系抗生物質分析法の複数機関による評価

Inter-laboratory validation studies were conducted in 5 laboratories to validate the biological method for determination of tetracyclines in royal jelly. Oxytetracycline spiked at the levels of 0.2 and 1.0 ppm was analyzed. Mean recoveries were 88 and 90%, reproducibility relative standard deviations (RSD(R)) were 13.7 and 7.8%, and HORRAT(R) values were 0.7 and 0.5. Samples containing residues at the levels of 0.25 and 0.80 ppm were analyzed. Mean recoveries were 73 and 77%, RSD(R) were 12.6 and 10.5%, and HORRAT(R) values were 0.6 and 0.6. The determination limit was 0.1 ppm (oxytetracycline, tetracycline) and 0.02 ppm (chlortetracycline). These results show that this method has good performance.

Field method for the micro-quantitative determination of tetracycline in human urine

A simple, fast and sensitive titrimetric method has been developed for the determination of tetracycline hydrochloride in human urine using m-dinitrobenzene as color agent and Dowex 1 × 8 as detection medium.

Liquid Chromatographic Determination of Tetracycline Residues in Animal Feeds

A liquid chromatographic method for the multiresidue determination of tetracyclines (TCs) in feeds is described. The levels of quantitation were 10 ppm each for tetracycline-HCl (TC), oxytetracycline (OTC), and chlortetracycline-HCl (CTC); the detection limit was 40 ppb for each. The calibration curves were linear between 2.5 and 100 ppm. The procedure involved double extraction with pH 2.0 and pH 4.5 McIlvain buffers, cleanup on a Sephadex LH-20 column, separation on a Nova-Pak C18 column, and detection at 370 nm. Recoveries of 10 micrograms/g of each TC in multiresidue feed samples ranged from 55.8 to 75.5% for OTC, 71.6 to 100% for TC, and 22.4 to 60.6% for CTC. The identities of the TCs were confirmed by thin layer chromatography.

New Colorimetric Method for Determination of Tetracycline and Oxytetracycline Via Charge Transfer Complex Formation

Abstract A colorimetric method for the determination of tetracycline and oxytetracycline is developed. It depends on nitration and reaction of the nitroderivatives with acetone in the presence of potassium hydroxide. A highly colored complex is produced. Variables such as temperature) time of heating, volume of nitrating mixture, acetone and potassium hydroxide have been evaluated to permit the selection of the most advantageous technique. Beer's Law is obeyed over the concentration range from 0.004 – 0.04 mg ml for tetracycline and oxytetracycline.