ABSTRACT
A new method for the characterization and determination of crystalline miokamycin in a pharmaceutical preparation using near infrared spectroscopy is proposed. The procedure involves the identification against a spectral library of samples with contents above the legal limit (viz. 5% of the crystalline form in amorphous miokamycin) using the Mahalanobis distance as the discriminating criterion, and the determination of the total and crystalline contents using partial least squares models that provide errors <1%. The proposed method was validated following the ICH guidelines. Its simplicity and expeditiousness make it highly suitable for quality control analyses of solid dosage forms.
Subject(s)
Miocamycin/analysis , Chemistry, Pharmaceutical , Crystallization , Guidelines as Topic , Powders , Quality Control , Reference Standards , Spectroscopy, Near-Infrared/instrumentation , Spectroscopy, Near-Infrared/methods , X-Ray DiffractionABSTRACT
A fast gradient reversed-phase liquid chromatography (LC) method, using an acetonitrile gradient was developed to determine the chromatographic hydrophobicity index (CHI), as reported by Valco et al. (Anal. Chem. 1997, 69, 2022-2029). The analytical method provides retention times, based on UV detection at two different wavelengths, which then are converted into CHI values after calibration with a set of test compounds. The CHI of each compound is measured at three different pH values, 2.0, 7.4, and 10.5; so using an 8-min gradient at each pH value one compound can be analyzed in approximately 24 min. The aim of this work is to improve the throughput of the CHI screening using a LC/MS approach, so the application of the LC/MS technique is an extension of the LC/UV approach previously reported by Valco et al. This approach allows contemporary injection of N compounds into the LC/MS system, the retention time of each compound can be then extracted from the selected ion recording chromatograms. The throughput of the existing screening method could be increased by N times, where N is the number of compounds injected, so only three runs are needed to determine the CHI at three different pH values for a set of N compounds. The highest value of N depends on the total number of channels that can be monitored simultaneously; in the present work, 32 channels were used. This LC/MS method has been tested for a number of commercial products analyzed as mixtures, and data obtained were compared with those coming from the classical LC/UV approach. In the same way, the method was tested for a number of compounds associated with two GlaxoWellcome projects in the antibacterial area. Data reported show that the LC/MS method can be successfully applied for analyzing compounds in mixtures and for compounds with poor UW absorption, which cannot be analyzed with the standard LC/UV method.
Subject(s)
Anti-Bacterial Agents/analysis , Chromatography, Liquid , Enzyme Inhibitors/analysis , Mass Spectrometry , Miocamycin/analogs & derivatives , Miocamycin/analysis , Enzyme Inhibitors/pharmacology , Topoisomerase II InhibitorsABSTRACT
A method for the characterization and determination of the crystalline from present in the amorphous miokamycin of a pharmaceutical preparation using near-infrared spectroscopy (NIRS) is proposed. The study leading to the development of the proposed method involved both qualitative (classification by residual variance analysis) and quantitative aspects (the determination of both total miokamycin and its crystalline form using multivariate calibration). Samples containing less than 5% of the total amount of miokamycin in its crystalline form can be accurately classified. Partial least-squares regression (PLSR) allows low contents of the crystalline form to be determined with absolute errors less than 1.5%, which is comparable to the limit of detection of the X-ray diffraction technique used as reference. Also, the total miokamycin content is determined by NIRS with errors less than 1%. The simplicity, expeditiousness and robustness of the proposed method make it an effective tool for determining both total and crystalline miokamycin in solid dosage forms.
Subject(s)
Miocamycin/analysis , Least-Squares Analysis , Spectroscopy, Near-InfraredABSTRACT
New methods for the determination of the nominal content of miokamycin in three commercial pharmaceutical preparations available in many different forms are proposed. Solid samples, grinding of which is the sole pretreatment required, are analysed by near infrared (NIR) spectroscopy, using a fibre-optic probe. The active principle is quantified by partial least-squares regression (PLSR). The three proposed methods were validated with a view to their use as control methods; the selectivity of the method, and the repeatability, intermediate precision, accuracy, linearity and robustness of each PLSR calibration model used were determined. The relative standard error of prediction (RSEP) was < 1.5% and the validation results testify to the suitability of the proposed methods.
Subject(s)
Anti-Bacterial Agents/analysis , Miocamycin/analysis , Pharmaceutical Preparations/chemistry , Spectroscopy, Near-InfraredABSTRACT
Fundamental studies in tissue drug extraction have been made using rokitamycin as a test probe. Radiolabelled rokitamycin was administered intravenously to 12 rats. Lungs and femurs were excised a few minutes later. Rokitamycin was extracted from lungs using six procedures (based either on ion-pairing, dissolution or deproteinization) whose performances (mean recovery and reproducibility) were compared. Three grinding procedures were also compared for the extraction of rokitamycin from bone: pulverization by a magnetic stirring bar in a liquid nitrogen bath, slicing into small pieces and crushing with pestle and mortar. The effect of binding proteins (albumin or alpha 1-acid glycoprotein) in the extraction mixture was also evaluated. Magnetic stirring bar grinding was the most efficient. Deproteinization was necessary to obtain the highest recovery, but the agent had to be chosen carefully. Binding proteins either had no effect or decreased the recovery of rokitamycin. Recovery from bone was lower than that from lung. Binding to cellular components in the post-extraction pellet was only 3% (lung) and 9% (bone). It is concluded that a careful optimization of the extraction procedure of a drug from a tissue allows quantitative and reproducible measurement of its concentration.
Subject(s)
Anti-Bacterial Agents/analysis , Bone and Bones/chemistry , Chemistry Techniques, Analytical/methods , Lung/chemistry , Miocamycin/analogs & derivatives , Animals , Carbon Radioisotopes , Chemical Phenomena , Chemistry, Physical , Male , Miocamycin/analysis , RatsABSTRACT
A group of 18 patients, before undergoing otoplastic surgery, was treated with miocamycin 600 mg in a single dose; middle ear tissue samples were collected at 2, 3, 4 and 6 h. A group of 20 patients suffering from secretory otitis media (SOM) was also treated with 600 mg of miocamycin in a single dose; collections of middle ear fluid (MEF) were performed at 1, 2, 4 and 6 h. Blood withdrawals from all patients occurred at the same time intervals. The miocamycin determination was performed by means of a microbiological method employing Sarcina lutea ATCC 9341. The highest serum levels (1.06 +/- 0.13 mg/l) were found at 2 h in the group from which middle ear mucosa had been withdrawn and at 1 h (2.2 +/- 0.5 mg/l) in the group from which the MEF had been collected; at 6 h miocamycin could be determined in both groups, with values ranging from 0.1 to 0.2 mg/l. In middle ear mucosa the miocamycin concentration was 1.52 +/- 0.27 mg/kg at 2 h and 0.2 +/- 0.09 mg/kg at 6 h. In MEF, the miocamycin concentration was 2.1 +/- 0.27 mg/l at 1 h and 0.24 +/- 0.05 mg/l at 6 h. The miocamycin concentration determined at 1 h in MEF was virtually equal to that in serum at the same time. At the following experimental times of withdrawal, the miocamycin concentration in mucosa specimens, as well as in MEF samples, proved to be markedly higher than values simultaneously found in serum.