Assay and purity control of oxytetracycline and doxycycline by thin-layer chromatography — a comparison with liquid chromatography

doi:10.1016/0731-7085(90)80138-F

Journal of Pharmaceutical and Biomedical Analysis

Volume 8, Issues 8–12, 1990, Pages 891-898

https://doi.org/10.1016/0731-7085(90)80138-F Get rights and content

Abstract

A thin-layer chromatographic (TLC) method using densitometry is described for the assay and purity control of oxytetracycline and doxycycline. With a mobile phase of dichloromethane—methanol—water (59:35:6, v/v/v) and a silica gel thin-layer, previously sprayed with 10% sodium edetate solution adjusted to pH 9.0, all the potential impurities of oxytetracycline or doxycycline are well separated from the main components and from each other. Results obtained with TLC are compared with those obtained by previously established liquid chromatography (LC) methods using poly(styrene-divinylbenzene) stationary phases. A good correlation was obtained (r > 0.9999). For TLC the relative standard deviation (RSD) for the assay of the main component was <2%, for LC the RSD was <1%.

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Sulfur and nitrogen binary doped carbon dots derived from ammonium thiocyanate for selective probing doxycycline in living cells and multicolor cell imaging
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Consequently, developing efficiently methods for the detection of doxycycline concentration is of great importance in pharmaceutical preparations, food safety and environmental monitoring. In the past 20 years, numerous analytical techniques including chromatography [3-5], capillary electrophoresis [6], spectrophotometry [7,8], fluorescence [9], electrochemistry [10,11], coupled technique [12-16] were used for doxycycline monitoring. However, these methods commonly suffered from low sensitivity and/or complicated preparation.
A novel sulfur and nitrogen binary doped carbon dots (S,N-CDs) was synthesized by one-step manner through the hydrothermal treatment of citric acid (CA) and ammonium thiocyanate, and the procedures for biomedical applications, including probing doxycycline in living cells and multicolor cell imaging were developed. The obtained S,N-CDs are stable in aqueous solution, possess a very high quantum yield (QY, 74.15%) and good photostability. The fluorescence of S,N-CDs can be specifically quenched by doxycycline, providing a convenient turn-off assay of doxycycline. This assay shows a wide linear detection range from 0.08 to 60 μM with a low detection limit of 20 nM. The present method also displays a good selectivity. More importantly, the S,N-CDs have an excellent biocompatibility and low cytotoxicity, allowing the multicolor cell imaging and doxycycline detection in living cells. Consequently, the developed doxycycline methods is facile, low-cost, biocompatible, sensitive and selective, which may hold the potential applications in the fields of food safety and environmental monitoring, as well as cancer therapy and related mechanism research.
Synergistic interactions between doxycycline and terpenic components of essential oils encapsulated within lipid nanocapsules against gram negative bacteria
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Doxycycline is useful in the treatment of respiratory tract infections because of its activity against intracellular atypical microorganisms known to be resistant to antibiotics (Brunton et al., 2006), such as β-lactams. Doxycycline has an excellent oral absorption and a satisfying half-life (Naidong et al.,1990). Its bacteriostatic activity and its rapid resistance when used alone explain why doxycycline is frequently given with other antibiotics like neomycine (Bryskier, 2005), ofloxacin, amoxicilline or antibacterial compounds like lysosomotropic agents (drugs capable of penetrating lysosomes) (Raoult et al., 1990).
The combination of essential oils (EOs) with antibiotics provides a promising strategy towards combating resistant bacteria. We have selected a mixture of 3 major components extracted from EOs: carvacrol (oregano oil), eugenol (clove oil) and cinnamaldehyde (cinnamon oil). These compounds were successfully encapsulated within lipid nanocapsules (LNCs). The EOs-loaded LNCs were characterised by a noticeably high drug loading of 20% and a very small particle diameter of 114 nm. The in vitro interactions between EOs-loaded LNCs and doxycycline were examined via checkerboard titration and time-kill assay against 5 Gram-negative strains: Acinetobacter baumannii SAN, A. baumannii RCH, Klebsiella pneumoniae, Escherichia coli and Pseudomonas aeruginosa. No growth inhibition interactions were found between EOs-loaded LNCs and doxycycline (FIC index between 0.7 and 1.30). However, when bactericidal effects were considered, a synergistic interaction was observed (FBC index equal to 0.5) against all tested strains. A synergistic effect was also observed in time-kill assay (a difference of at least 3 log between the combination and the most active agent alone). Scanning electron microscopy (SEM) was used to visualise the changes in the bacterial membrane. The holes in bacterial envelope and leakage of cellular contents were observed in SE micrographs after exposure to the EOs–LNCs and the doxycycline combination.
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2004, Journal of Chromatography A
A simple, rapid and sensitive reversed-phase high-performance liquid chromatographic (HPLC) method for the measurement of doxycycline concentrations in both drug delivery systems (DDS) and serum extracted from mice after intraperitoneal (free drug) and intravenous (doxycycline administered in DDS) administration, has been developed. For the analysis of doxycycline in DDS, a known amount of particles was dissolved in chloroform and, after precipitating the polymer with methanol, the drug was assessed in the supernatant. For doxycycline quantification in microsamples of serum, proteins were precipitated with acetonitrile before chromatographic analysis. After centrifugation, the supernatant was mixed with a mixture of methanol and acetic acid (1:1) for analysis. The samples were chromatographed on a narrow-bore C₁₈ column (Alltech Alltima $150 mm ×2.1$ mm) using a mobile phase with 55% acetic acid (5%), 25% acetonitrile and 20% methanol. Doxycycline was detected 347 nm and the run time was 10 min. Linearity was confirmed in the concentration range 0.4–80 μg/ml for doxycycline quantification in serum and from 1 to 800 μg/ml for doxycycline extracted from DDS samples.
Quantitative analysis of oxytetracycline and its impurities by LC-MS-MS
2004, Journal of Pharmaceutical and Biomedical Analysis
A liquid chromatographic-tandem mass spectrometric method using an Xterra MS C₁₈ chromatographic column ( $100 mm ×2.1$ mm i.d., 3.5 μm) that allows complete separation of oxytetracycline (OTC) and the impurities: 4-epi-oxytetracycline (EOTC), tetracycline (TC), 4-epi-tetracycline (ETC), 2-acetyl-2-decarboxamido-oxytetracycline (ADOTC), α-apo-oxytetracycline (α-AOTC) and β-apo-oxytetracycline (β-AOTC) was developed. Gradient elution was used and calibration curves were obtained using the scan mode selected reaction monitoring (SRM). Acceptable correlations were obtained for OTC, TC, EOTC and ADOTC whereas the correlations of α-AOTC and β-AOTC were less accurate resulting in higher limits of quantification (LOQ) and limits of detection (LOD) relative to the other compounds. The intraday and interday accuracy varied for all the compounds from 90 to 112% and the intraday and interday precision were lower than 7.1%. The method was applied for analysis of commercial available ointments containing OTC resulting in an acceptable quantification of OTC and the impurities in the drug preparations. The advantage of this method compared to the other separation methods is an empty separation window right after the large peak corresponding to OTC in the chromatogram, which facilitates an accurate determination of ADOTC and the other impurities.
Assay and purity control of tetracycline, chlortetracycline and oxytetracycline in animal feeds and premixes by TLC densitometry with fluorescence detection
2003, Journal of Pharmaceutical and Biomedical Analysis
Methods using TLC densitometry with fluorescence detection are described for the assay and purity control of tetracycline (TC), chlortetracycline (CTC), and oxytetracycline (OTC) in animal feeds and premixes. With a silica gel layer previously sprayed with 10% (m/v) sodium EDTA solution adjusted to pH 8.0 or 9.0, all the major impurities were separated from the main components and from each other. The mobile phase consisted of dichloromethane, methanol, and water. After development, the plate was dipped in a 30% (v/v) solution of liquid paraffin in hexane. Quantitation was realized by fluorescence densitometry at 400 nm. The limit of quantitation (LOQ) for tetracycline impurities is 0.8 μg/g, corresponding to 0.2% of the label claimed tetracycline (400 μg/g). The LOQ for impurities of tetracycline and chlortetracycline in premixes is 0.2% of the label-claimed TC (40 mg/g) and CTC (200 or 400 mg/g). The LOQ for impurities of oxytetracycline in a premix is 0.1% of the label claimed OTC (100 mg/g).

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^☆: Presented at the Second International Symposium on “Pharmaceutical and Biomedical Analysis”, April 1990, York, UK.

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Assay and purity control of oxytetracycline and doxycycline by thin-layer chromatography — a comparison with liquid chromatography☆

Abstract

J. Chromatogr.

J. Pharm. Biomed. Anal.

J. Pharm. Sci.

J. Pharm. Sci.