Isolation of doxycycline, 6-epidoxycycline and 2-acetyl-2- decarboxamidometacycline from commercial metacycline by preparative column liquid chromatography on silica gel.

Isolation of doxycycline, 6-epidoxycycline and 2-acetyl-2-decaboxamidometacycline from commercial metacycline was achieved by preparative column liquid chromatography on silica gel, previously impregnated with edetate (EDTA). Careful control of the pH of EDTA allowed fine tuning of the separation. The mobile phases were composed of dichloromethane, methanol and .1 mM EDTA at pH 9.0 or 6.0. Structures were confirmed with nuclear magnetic resonance spectroscopy. The presence of doxycycline and its 6-epimer in commercial metacycline has not previously been described. The presence of the 2-acetyl derivative was not surprising since analogous 2-acetyl derivatives have been identified in other tetracyclines.     

Determination of Doxycycline, 4-epidoxycycline,and 6-epidoxycycline in Aquatic Animal Muscle Tissue by an Optimized Extraction Protocol and Ultra-performance Performance Liquid Chromatography with Ultraviolet Detection

A fast ultra-performance liquid chromatography method was developed for the separation and quantification of doxycycline, 4-epidoxycycline, and 6-epidoxycycline in the muscles of channel catfish, shrimp, eel and turtle. The analytes were separated by comprehensive optimization of the chromatographic conditions. The extraction method was performed using a modified QuEChERS methodology. 2?g of muscle sample were mixed with 5?mL Mcllvaine buffer (pH 4), and 4?g of sodium sulfate and 2?g of sodium chloride were added for liquid–liquid partitioning. The mixture was agitated for 30?s and extracted with 10?mL of acidified acetonitrile. The resulting extract was purified using 200?mg of a C₁₈-silica adsorbent. The samples were analyzed using ultra-performance liquid chromatography with ultraviolet detection using a BEH C18 column (2.1?mm ×100?mm, 1.7?μm). The limits of detection were less than 25?μg/kg and the limits of quantitation were less than 50?μg/kg for all analytes. The recoveries of the analytes were from 62.3% to 89.0% with relative standard derivations below 10.0%. The method provided efficient extraction and purification that enabled rugged and low-cost determination of the analytes.     

Specific feature of water vapor sorption by chitosan medicated films

Sorption properties of chitosan-based medicated films were studied. IR and UV spectroscopies were used to confirm that chitosan chemically interacts with medicinal preparations. It is assumed that structural changes occurring in the polymer matrix either as a result of introduction of medicinal substances, or in the course of isothermal annealing of films lead to significant departures of the fundamental aspects of the sorption process from the classical Fick’s mechanism.     

Determination of emamectin benzoate in medicated fish feed: a multisite study

A new method was developed for the quantitation of emamectin benzoate in medicated fish feed at levels between 1 and 30 ppm. The new procedure, based on a previously reported assay, consists of a wet methanolic extraction of ground feed, followed by solid-phase extraction and injection onto a gradient liquid chromatographic system. A multisite study involving 3 laboratories (the developing laboratory and 2 independent laboratories) was performed to evaluate precision, recovery, linearity, and sensitivity. Mean recove;ries for triplicate analyses at 3 levels, performed by 2 analysts per laboratory, were between 89 and 97%, with coefficients of variation ranging from 1.6 to 8.6%. Coefficients of determination (r2) obtained from the plotted data were > or =0.993. The precision of the method, determined from 6 replicate preparations from the same batch of medicated feed assayed in 3 separate trials per laboratory, was between 0.6 and 5.8%. The quantitation limit was established at 0.5 ppm. Specificity and robustness studies were performed by the developing laboratory.     

Ab initio study of 2,4-substituted azolidines. II. Amino-imino tautomerism of 2-aminothiazolidine-4-one and 4-aminothiazolidine-2-one in water solution

The relative stabilities of the tautomers of 2-aminothiazolidine-4-one and 4-aminothiazolidine-2-one were calculated at the MP2/6-31+G(d,p) level by considering their mono- and trihydrated complexes. Single-point calculations at the MP4/6-31+G(d,p)//MP2/6-31+G(d,p) level of theory were performed to obtain more accurate energies. The values of proton transfer barriers in the isolated, mono- and trihydrated tautomers of 2-aminothiazolidine-4-one (2AT) and 4-aminothiazolidine-2-one (4AT) were calculated for two different mechanisms of tautomerisation. In the absence of water, the process of proton transfer should not occur. Addition of water molecules decreases the barrier making the process faster, as the participation of two water molecules in a proton transfer reaction is more favorable than the participation of only one water molecule. To estimate the effect of the medium (water) on the relative stabilities of the tautomers of the studied compounds we applied the polarizable continuum model (PCM). (13)C NMR chemical shieldings were calculated using the GIAO approach at MP2/6-31+G(d,p) optimized geometries. HF and the DFT B3LYP functional with 6-31+G(d,p) basis set were employed. The quantum chemical results for the chemical shifts in gas phase and in polar solvents (water and DMSO) were compared with experimental data. TD DFT B3LYP/aug-cc-pVTZ calculations were performed to predict the absorption maxima of tautomers A and B of 2AT and 4AT.     

Residue determination of glyphosate in environmental water samples with high-performance liquid chromatography and UV detection after derivatization with 4-chloro-3,5-dinitrobenzotrifluoride

A pre-column derivatization high-performance liquid chromatographic method for glyphosate analysis has been developed. Derivatization of glyphosate was performed with 4-chloro-3,5-dinitrobenzotrifluoride (CNBF). In pH 9.5 H₃BO₃-Na₂B₄O₇ media, the reaction of glyphosate with CNBF completed at 60 °C for 30 min. The labeled glyphosate was separated on a Kromasil C18 column (250 mm × 4.6 mm, 5 μm) at room temperature and UV detection was applied at 360 nm. The separation of labeled glyphosate was achieved within 15 min by gradient elution mode. Compared to other pre-column derivatization, this derivatization was performed more mildly, the derivative was more stable, and the detection limits of a few reagents were higher than CNBF, except 9-fluorenylmethyl chloroformate (FMOC-Cl) using fluorescence and mass spectrometry, however, this reagent avoid to be removed after derivatization like FMOC-Cl. The detection limit of glyphosate was 0.009 mg L⁻¹ (S/N = 3) without preconcentration and reach MRL, which is set at the level of 0.1 mg L⁻¹ in China. The method linearity correlation coefficient was 0.9999, in concentrations ranging from 0.3 to 48.5 mg L⁻¹. The proposed method has been applied to the quantitative determination of glyphosate in environmental water with recoveries of 91.80-100.20% and R.S.D. of 2.27-6.80, depending on the sample investigated.     

Chemometric study of liquid water simulations. I. The parameters of the TIP4P model potential

The multivariate chemometric techniques two level factorial design (TLFD) and principal component analysis (PCA) were used to investigate the TIP4P model potential behavior with respect to perturbations on all intermolecular interaction parameters. The effects of these perturbations were calculated for the enthalpy of vaporization, the density, the first maximum of the radial distribution functions of the O-H and O-O pairs, and the second maximum of the radial distribution function of the O-H pair obtained from Monte Carlo simulations of liquid water at 25 degrees C. The principal effects were quantified and rationalized in terms of the pair-wise interaction potential of the TIP4P model. They also corroborate previously published sensitivity analysis results using molecular dynamics and other model potentials. In addition, significant interaction effects between some parameters of the TIP4P model potential were observed and quantified, which hardly could be obtained without such a statistic approach. These interaction effects are very regular and systematic, and their behavior has not been encountered in other chemometric studies and cannot be rationalized in terms of the functional form of the pair-wise potential.     

Comparison study of two procedures for the determination of emamectin benzoate in medicated fish feed

A new method has been developed for the determination of emamectin benzoate in fish feed. The method uses a wet extraction, cleanup by solid-phase extraction, and quantitation and separation by liquid chromatography (LC). In this paper, we compare the performance of this method with that of a previously reported LC assay for the determination of emamectin benzoate in fish feed. Although similar to the previous method, the new procedure uses a different sample pretreatment, wet extraction, and quantitation method. The performance of the new method was compared with that of the previously reported method by analyses of 22 medicated feed samples from various commercial sources. A comparison of the results presented here reveals slightly lower assay values obtained with the new method. Although a paired sample t-test indicates the difference in results is significant, this difference is within the method precision of either procedure.     

Exendin-4中13号残基的分子动力学模拟

Exendin-4作为胰腺GLP-1受体上的一种有效的激活剂, 是一种含有39个氨基酸残基的多肽, 其第13号氨基酸Gln突变为Tyr, 使活性增强. 应用分子动力学模拟方法, 分别优化了突变前后, Exendin-4与蛋白的复合物结构, 并对整体结构的性质、静电势、相互作用模式及能量进行了分析. 阐明了Gln突变为Tyr的活性增强的内在原因, 结果表明, 突变的Exendin-4能够通过改变自身结构的局部柔性调整与蛋白受体相互作用, 从而可以改善Exendin-4与其蛋白受体的结合能力.     

Photochemistry of 4-Pyrones in water. Formation of dihydroxycyclopentenones and furan derivatives

Irradiation of aqueous solutions of aikyl substituted 4-pyrones leads to the formation of dihydroxycyclopentenones. These primary products are also photolabile and in certain cases are photochemically converted to furan derivatives.     

Determination of emamectin benzoate in medicated fish feed

A method was developed to quantitate emamectin benzoate in fish feed at levels between 5 and 15 ppm. The active ingredient is extracted from 20 g medicated feed into aqueous-methanolic solvent by overnight shaking. A solid-phase extraction procedure using a 2 g C18 cartridge is then used to concentrate the active residue and remove interfering matrix components. The extracted drug and internal standard are eluted from the cartridge, evaporated to dryness, and reconstituted in methanol. A control feed sample and fortified control working standard are simultaneously prepared. Remaining interferences and sample analysis are further separated on a gradient liquid chromatographic system. Recovery of emamectin benzoate from fortified feeds ranged from 97 to 100%, with a coefficient of variation (CV) of 1.2%. Determination of emamectin benzoate in medicated feeds resulted in CVs ranging from 2.3 to 4.2% and recoveries of 88 to 98% of label claim.     

Identification of electrical degradation products of 4-chlorophenol in water.

The electrical decomposition of 4-chlorophenol in water was examined with iridium dioxide doped on atitanium electrode. A number of electrical degradation products of 4-chlorophenol, such as hydroquinone and chlorohydroquinone via the addition of hydroxyl radicals, and dichlorophenol through addition of chlorine radical, were observed as major products. Moreover, hydroxylated chlorobiphenylethers, hydroxylated dibenzo-p-dioxin/furans and hydroxylated chlorobiphenyls formed by a dimerization process during the electrolysis process of 4-chlorophenol were also observed. On the other hand, benzoquinone, muconic acid and aldehyde derivatives that were further oxidative products of hydroquinone formed by photocatalysis process, were not observed. The electrical decomposition products of 4-chlorophenol were trimethylsilylated and then identified by gas chromatography-mass spectrometry. The degradation rate of 4-chlorophenol in water by iridium oxide electrode was measured against the electrical process duration. After iridium electrical process for 120 min, about 50% of 4-chlorophenol was converted into a number of products through oxidation processes. On the basis of the identified products, the degradation pathways of 4-chlorophenol under electrolysis process were proposed.     

Acrylate terpolymer in fabrication of medicated skin patches

An acrylate based pressure sensitive adhesive (PSA) was synthesized to design a drug‐in‐adhesive (DIA) type transdermal therapeutic system (TTS) for nitroglycerin used in the treatment of angina pectoris. 2‐Ethylhexyl acrylate (EHA), methyl methacrylate (MMA) and acrylic acid (AA) were used to synthesize the PSA by free radical solution polymerization. The effects of reaction time, reaction temperature, initiator concentration and solvent on polymerization were studied. The synthesized terpolymer was characterized by ¹H‐NMR, FT‐IR, differential scanning calorimetry (DSC) and gel permeation chromatography (GPC) and also evaluated for intrinsic viscosity, refractive index, peel strength, moisture uptake and skin irritation potential. The PSA was used to develop DIA type patches of nitroglycerin. The patches were cast using solvent evaporation technique and dried at controlled temperature. The patches were evaluated for thickness uniformity, weight variation, peel strength and moisture pick‐up. The percent drug content and in vitro drug release was determined by high pressure liquid chromatography (HPLC) method. On the basis of in vitro release profile, patches were selected for in vitro skin permeation studies. The developed formulation TP‐1 (K = 24.892 mcg/cm²/hr) followed zero‐order rate kinetics and showed better skin permeation rate in comparison to the marketed TTS (MTTS) (K = 17.413 mcg/cm²/hr). TP‐1 was subjected to stability testing for a period of 1 year according to ICH guidelines. The patches were found to be stable and an expiry date of 2 years was predicted with storage at 25 °C or below. Copyright © 2001 John Wiley & Sons, Ltd.     

Carbazole as fluorescence carrier for preparation of doxycycline sensor.

Carbazole is a promising pharmaceutical species. A novel optical sensor for determining doxycycline based on the fluorescence quenching of N-allylcarbazole immobilized on an quartz glass plate surface by covalent bonding has been described. The sensor shows satisfactory virtues in reversibility, repeatability, selectivity and sufficient lifetime resulting from its excellent optode membrane. Its response time is less than 60 s. The determination range and detection limit of the sensor are 6.0 x 10(-7)-2.0 x 10(-3) M and 2.0 x 10(-7) M, respectively. The lifetime of each sensor is at least three to four months. The sensor can be used for direct determination of doxycycline in pharmaceutical preparations and urine samples.     

High-performance liquid chromatographic analysis of oxytetracycline in chinook salmon following administration of medicated feed.

A high-performance liquid chromatographic assay was developed to detect oxytetracycline (OTC) in chinook salmon muscle tissue. A solid-phase extraction protocol was used to recover OTC and the internal standard, epitetracycline hydrochloride, from the salmon tissue samples. OTC was analyzed using a mobile phase of methanol-0.02 M phosphate buffer, pH 2.25 (60:190), an ultraviolet detection wavelength of 365 nm and 250 mm x 4.6 mm I.D. Ultrasphere ODS column. A linear calibration curve (r2 = 0.999) of OTC in salmon muscle tissue from 0.05 to 3.0 ppm was obtained. Using a signal-to-noise ratio of 5:1, the OTC detection limit was 0.5 ppm in salmon muscle tissue. OTC recovery (74.4%) and intra-assay variability (2.3%) were optimized for salmon muscle tissue. An in vivo feeding study was performed by administrating OTC-medicated feed for a period of 10 days, followed by a 42-day sampling period. The half-life for the elimination of OTC in chinook salmon muscle tissue was found to be 5.4 days.     

Liquid chromatographic determination of pyrantel tartrate in medicated formulations

The validation of a novel liquid chromatographic (LC) method for the determination of pyrantel tartrate in feed is presented. The method provides a significant improvement over the efficiency and precision of AOAC Official Method 978.30. The method was shown to be accurate, precise, linear, and robust for medicated articles. Unlike the official method, the LC method was shown to be a superior stability-indicating method. After the method was validated by using laboratory blends, the effectiveness of the method was demonstrated with marketed product as well.     

Microcolumn high-performance liquid chromatographic assay for doxycycline in isolated alveolar macrophages.

A procedure for the isolation of doxycycline from the alveolar macrophages is described. Due to the minimal amount of the sample, it was necessary to employ microcolumn HPLC. A great advantage of the methodology developed in the present paper consists in the direct injection of an aqueous supernatant over disintegrated macrophages without the use of liquid-liquid or solid-liquid extraction. Another advantage of the method is the fact that the injection of an aqueous solution on a microcolumn with a lipophilic stationary phase makes the analyzed substances concentrate, which increases the sensitivity of assay. Separation was performed on a Separon SGX C18 microcolumn, 150 x 1 mm, detection at 254 nm by UV, 1 microliter flow cell. Rabbits were treated intravenously with Vibramycin inj. sic. in a dose of 13 mg/kg body weight doxycycline. A procedure for washing macrophages from rabbit lungs was developed. Dry matter of macrophages ranged from 5.9 to 11.4 mg. In the samples of three rabbits the levels of doxycycline ranged from 110 to 270 ng per 1 mg of macrophages dry matter.