[Determination of antibiotics residues in the raw fresh milk of farms in the Miyun County of Beijing with Mg2+ -sensitized metacycline fluorescence microscopic imaging technique].

Mg(2+) -sensitized metacycline fluorescence microscopic imaging technique was applied to detect the raw fresh milk of four cows breeding farms in the Miyun County of Beijing based on the capillary effect of solvent on solid supports. In the presence of NH3-NH4 Cl buffer solution (pH 9.99) and PVA-124, Mg2+ and metacycline can form a strong fluorescence complex of 1 : 1, and Mg(2+) -metacycline complex can form an SOR on the hydrophobic supports with the diameter of 0.93 mm and its ring belt width of 26.2 microm. By measuring the fluorescence intensity of the ring, the quantitative analysis of metacycline was achieved with the detection limit (3sigma) of 8.8 x 10(-14) mol x ring(-1) (1.8 x 10(-7) mol x L(-1)) and linear range of 2.2 x 10(-13) -3.6 x 10(-12) mol x ring(-1) (4.4 x 10(-7) -7.2 x 10(-6) mol x L(-1)) when 0.50 microL droplet was spotted. This method has been satisfactorily applied to the determination of metacycline in the raw fresh milk samples with the recovery of 93.8%-108% and RSD less than 4.3%.

Determination of tetracyclines residues in honey by on-line solid-phase extraction high-performance liquid chromatography.

An automated system using on-line solid-phase extraction (SPE) high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection was developed for the determination of tetracyclines (TCs), such as tetracycline (TC), oxytetracycline (OTC), chlortetracycline (CTC), metacycline (MC), and doxycycline (DC) in honey. One milliliter diluted honey sample was injected into a conditioned C18 SPE column and the matrix was washed out with water for 3 min. By rotation of the switching valve, TCs were eluted and transferred to the analytical column by the chromatographic mobile phase. Chromatographic conditions were optimized. TCs were separated in less than 8 min with a gradient elution using a mixture of 0.8% formic acid and acetonitrile. The UV detection was performed at 365 nm. The conditions for on-line SPE, including solvent and total time for loading sample and washing matrix were also optimized. Time for extraction and separation decreased greatly. For the five kinds of TCs, the limits of detection (LODs) at a signal-to-noise of 3 ranged from 5 to 12 ng g(-1). The relative standard deviations (R.S.D.) for the determination of TCs ranged from 3.4 to 7.1% within a day and ranged from 3.2 to 8.9% in 3 days, respectively.

Quantitative analysis of methacycline hydrochloride by direct potentiometry using the internal solid contact sensor.

An internal solid contact sensor (ISCS) for the determination of methacycline hydrochloride (MC.Cl), Pt/PPy/PVC(MC-PT), is described, based on the use of conducting poly(pyrrole) (PPy) as solid contact material and MC-phosphotungstate (PT) as the ion exchanger and dibutyl phthalate (DBP) as the plasticizer. A direct potentiometric method for the quantitative analysis of MC.Cl is also described. Under the condition of pH 2.7, the linear concentration range, slope (25 degrees C) and detection limit of the sensor are 6.4 x 1.0(-6) - 3.0 x 1.0(-3) M, 52.4 +/- 0.2 mV/decade and 4.4 x 1.0(-6) M, respectively. The response time is <5 s. The determinations of MC.Cl in tablets were carried out by direct potentiometry. The average recovery and relative standard deviation are 100.1 and 0.7% (n = 4), respectively.

[Determination of metacycline in mixture sample by synchronous-derivative fluorimetry].

A new synchronous-derivative fluorimetric method for the determination of metacycline (MTC) in the presence of oxytetracycline (OTC) is proposed. The degradation products of MTC in 0.1 mol x L(-1) NaOH are strongly fluorescent. The addition of cetyltrimethylammonium bromide (CTMAB) enhances the fluorescence intensity 2-fold. The synchronous fluorescence spectra of MTC and OTC degradation products do not interfere with each other under the condition that deltalambda = 100 nm. MTC can be determined in the presence of OTC in the range of OTC+MTC ratio of 1+116-1+1. The recoveries of MTC in the mixture sample are in the range of 94%-102%.

Precision study on capillary electrophoresis methods for metacycline.

A CE method for metacycline (MTC) determination was investigated in an inter-laboratory experiment. Many problems were encountered in this study, most of which were related to the transfer of the method to different CE equipment. The reported problems could be classified into different categories: problems related to the precision, to the parameters in the protocol, and to the MTC peak shape. As the peak shape problem was partially responsible for the poor precision, a new CE method was developed in order to obtain a good MTC peak shape on all equipment. The precision of this new method for MTC determination was examined in an intermediate precision study, where the influence of the factors "time" and "equipment" was investigated. Although the new method could be transferred to different instruments, the precision remained poor mainly due to the contributions of the between-replicate and the between-injection variances.

Determination of tetracyclines residues in honey using high-performance liquid chromatography with potassium permanganate-sodium sulfite-beta-cyclodextrin chemiluminescence detection.

A novel method was developed for the simultaneous determination of tetracycline antibiotic (TCA) residues such as oxytetracycline (OTC), tetracycline (TC), and metacycline (MTC) by high-performance liquid chromatography (HPLC) coupled with chemiluminescence (CL) detection. The procedure was based on the chemiluminescent enhancement by TCAs of the potassium permanganate-sodium sulfite-beta-cyclodextrin system in a phosphoric acid medium. The separation was carried out with an isocratic elution using a mixture of acetonitrile and 0.001 M phosphoric acid. For the three TCAs, the detection limits at a signal-to-noise of 3 ranged from 0.9 to 5.0 ng/ml. The relative standard deviations for the determination of TCAs ranged from 3.1 to 7.4% within a day (n=11) and ranged from 2.2 to 8.6% in 3 days (n=9), respectively. The method was successfully applied to the determination of TCA residues in honey samples. The possible mechanism of the CL reaction was also discussed.

Improvements in europium sensitized fluorimetric determination of demeclocycline and methacycline.

Demeclocycline (DM) and methacycline (MT) have been determined by europium-sensitized fluorescence, using EDTA as co-ligand and cetyltrimethylammonium chloride as surfactant. The methods have been developed in slightly alkaline solutions, with the formation of a new chelate where the lanthanide ion is bound to the beta-diketone group. Calibration graphs between 0.01 and 0.1 microg mL(-1) have been obtained for DM and MT determination. Both methods have been applied to the determination of these tetracyclines in serum samples with satisfactory recovery results.

Analysis of metacycline by capillary electrophoresis.

The development and validation of an optimized capillary electrophoresis method for the determination of metacycline in the presence of its related substances by capillary electrophoresis is shown. The influence of methanol as organic modifier, buffer pH, buffer concentration, capillary length, column temperature, Triton X-100 and methyl-beta-cyclodextrin was investigated. A central composite design was performed in order to optimize the method. The optimal separation conditions were: uncoated fused-silica capillary (39 cm total length, 31 cm effective length, 50 microm ID); as background electrolyte a solution of 160 mM sodium carbonate and 1 mM EDTA (pH 10.35)/methanol (89:13 v/v); temperature, 15 degrees C; voltage, 12 kV. The method showed good selectivity, repeatability, linearity, and sensitivity. The limits of detection and quantitation are 0.024% and 0.06%, respectively, relative to a 2.5 mg/mL solution. Six commercial samples were analyzed quantitatively.

Evaluation of amperometric detection for the liquid-chromatographic determination of tetracycline antibiotics and their common contaminants in pharmaceutical formulations.

A sensitive high-performance liquid chromatographic technique with amperometric detection has been developed for the determination of seven commercially used tetracyclines in bulk powders and pharmaceutical preparations. The technique is based on the oxidation of these compounds and their contaminants at the glassy carbon electrode. The extraction procedures are simple and the HPLC conditions separate the tetracyclines from their major degradation products. The chromatography was performed using a commercially available octadecylsilane column, with a mobile phase: KH2 PO4 (pH = 2.5; 0.05 M) - acetonitrile (84:16, v/v) and detection at 1.2 V. The technique permits the simultaneous determination of trace amounts of chlortetracycline, demeclocycline, doxycycline, methacycline, minocycline, oxytetracycline and tetracycline as well as the separation of their common impurities (epi, anhydro and epianhydro contaminants) with detection limits of 0.1-1.0 ng microl(-1) and recoveries of 99.1-100.4%. No interference was observed from the commonly present excipients in pharmaceutical formulations.

Simple continuous and simultaneous determination of tetracycline residues.

A new continuous separation method was developed for the determination of five different tetracyclines (oxytetracycline, tetracycline, chlortetracycline, methacycline and doxycycline). A bioassay using minimum medium (MM) seeded with Bacillus subtilis ATCC 6633 was carried out for the detection, with simple extraction from the agar block of the clear inhibition zone on the MM produced by the mixed tetracyclines. The extract was subjected to continuous identification by high performance liquid chromatography using a mu-Bondapack C18 column. The tetracyclines were separated at ambient temperature using a mobile phase of 0.01 M oxalic acid: acetonitrile: N,N-dimethylformamide (74:18:8, v/v/v) at a flow rate of 1.0 ml min-1. A variable-wavelength detector set at 355 nm and recorder set at 4 mm min-1 were used for the detection. The entire mixture was resolved as five peaks with retention times ranging from 2.75 to 9.65 minutes. This continuous, simple and rapid method of detection, extraction and identification may be useful for routine laboratory testing of residual antimicrobial agents in food.

Determination of metacycline and related substances by column liquid chromatography on poly(styrene-divinylbenzene).

Isocratic column liquid chromatography on poly(styrene-divinylbenzene) copolymer allowed complete separation of metacycline, 4-epimetacycline, oxytetracycline, doxycycline and 6-epidoxycycline. 2-Acetyl-2-decarboxamidometacycline was eluted on the tail of metacycline. The mobile phase was 2-methyl-2-propanol-0.2 M phosphate buffer (pH 9.0)-0.01 M sodium ethylenediaminetetraacetate (pH 9.0)-water (2.5:10:10:77.5, m/v/v/v). The flow-rate was 1.0 ml/min and detection was performed at 254 nm. Official standards were compared and a number of commercial bulk samples and specialties were analysed. 2-Acetyl-2 decarboxamidometacycline, 6-epidoxycycline and doxycycline were the main impurities, while 4-epimetacycline and oxytetracycline were minor impurities.

[Fluorimetric study of methacycline complex formation with beryllium, aluminum, magnesium, calcium and zinc ions]. / Fluorimetricheskoe izuchenie kompleksoobrazovaniia metatsiklina s ionami berilliia, aliuminiia, magniia, kal'tsiia i tsinka.

Complex formation of metacycline (Pv) with beryllium, aluminium, magnesium, calcium and zinc ions in aqueous solutions was studied fluorometrically. The antibiotic formed the following compounds: CaPv2 (pH 9.45), MgPv (pH 10.40), Be Pv (pH 7.0 and 11), Ca Pv (pH 13.5), Zn Pv (pH 7.15) and Al Pv (pH 6.0). Relationship between the solution fluorescence level and the values of pH, the quantum yield of fluorescence of Pv and the complexes and the kinetics of the reactions of complex formation of Pv with Be and Al were studied. The mechanism of Pv complex formation with the metal ions is discussed. A method for analysis of microgram amounts of Pv in biological fluids was developed. The method is based on the antibiotic fluorescent reaction with magnesium ions.

A high performance liquid chromatographic system for the analysis of tetracycline drug standards, analogs, degradation products and other impurities.

This paper describes the high performance liquid chromatographic (HPLC) analysis of eight parent tetracycline standards: tetracycline, chlortetracycline, rolitetracycline, oxitetracycline, minocycline, doxycycline, democlocycline, methacycline; and three tetracycline epimers: epitetracycline, epianhydrotetracycline, and anhydrotetracycline. The HPLC system employs an octadecylsilane reverse phase column and an isopropanol-diethanolamine-phosphate-ammonium EDTA-water mobile phase. This system produced at least partial resolution of all eight parent compounds and many of their degradation products.

The parenteral application of a new metacycline preparation, tri-metacycline.

Tri-metacycline, one of the new tetracycline complexes (tritetracyclines), is prepared by mere dissolution of metacycline hydrochloride in an aqueous solution of the complexing agent. In vitro and in vivo studies show a high antibiotic activity. Significantly lower MIC values were found for tri-metacycline than for the parent compound. Parenteral administration resulted in high sera and tissue values, without signs of accumulation; excretion via the kidneys was proved.