Spectrophotometric determination of cimetidine in pharmaceuticals and urine using batch and flow-injection methods

Two sensitive and fast spectrophotometric methods using batch and flow-injection procedures for the determination of cimetidine (CMT) are proposed. The methods are based on the formation of a green complex between this drug and Cu(II) in acetic/acetate medium of pH 5.9. The calibration graphs resulting from measuring the absorbance at 330 nm are linear over the ranges 2.5 x 10(-6)-1.0 x 10(-3) and 5 x 10(-6)-2.0 x 10(-3) M with detection limits of 9.5 x 10(-7) and 2.1 x 10(-6) for batch and flow-injection methods, respectively. The methods are applied to the routine analysis of CMT in pharmaceuticals and human urine.     

Spectrophotometric and fluorimetric methods for the determination of meloxicam in dosage forms

Four simple and accurate methods are presented for the determination of meloxicam in dosage forms. These methods are based on: the direct measurements of the differential spectra at 339.9-384.7 nm (A), the 1D-values at 322-368 nm and 2D-values at 343.2-385.6 nm (B), the formation of an ion-association complex between the drug and safranin T with subsequent absorption measurement at 518 nm (C) and fluorescence measurement at 582 nm (D). All variables were studied to optimize the formation of the ion-association complex. Beer's law was valid over the concentration range 2-10 microg ml(-1) (method A), 1-10 microg ml(-1) (method B), 4.0-12 microg ml(-1) (method C) and 0.4-1.2 microg ml(-1) (method D). The detection limits were 0.11, 0.07, 0.10, 0.33 and 8.74 x 10(-3) microg ml(-1) for methods A, B, C and D, respectively. The proposed methods were successfully applied to the assay of meloxicam in tablets and suppositories. The procedures were rapid, simple and suitable for quality control applications.     

Fluorimetric determination of chloroxine using manual and flow-injection methods

A reliable and highly sensitive method is described for the determination of chloroxine in pharmaceutical preparations. It involves the formation of a complex between chloroxine and aluminum(III) in a micellar medium. The complex is a very fluorescent species, and there is a linear relationship between chloroxine concentration and fluorescence intensity over the range {ce:inline-formula}2.0 × 10 ⁻⁸ −5.1 × 10 ⁻⁵mol1 ⁻¹{/ce:inline-formula}. The limit of detection is {ce:inline-formula}5 × 10 ⁻⁹mol1 ⁻¹{/ce:inline-formula}. The method can be easily adapted to a flow system using a three-channel manifold, the peak height being proportional to the chloroxine concentration over the range {ce:inline-formula}5.6 × 10 ⁻⁷ − 5.6 × 10 ⁻⁵mol1 ⁻¹{/ce:inline-formula}. Manual and flow-injection procedures permit the determination of chloroxine in the presence of chlorquinaldol, and have been successfully applied to the determination of chloroxine in pharmaceutical preparations.     

Spectrophotometric determination of pefloxacin mesylate in pharmaceuticals

A spectrophotometric method is described for assay of pefloxacin mesylate (PFM) in bulk drug and in tablets. The method is based on back extraction of the bromophenol blue dye at pH 5.2 from the dye-drug ion pair followed by measurement of the dye absorbance at 590 nm. The working conditions of the method were investigated and optimized. Beer's law plot showed a good correlation in the concentration range of 0.15-1.25 microg mL(-1). Sensitivity indices such as molar absorptivity, limits of detection and quantification are reported. Intra-day and inter-day precision, and accuracy of the methods were established according to the ICH guidelines, and the er values were in the range of -1.7 to 1.8% with RSD values ranging from 1.0 to 1.1%. The method was successfully applied to the assay of PFM in tablet preparations with recoveries varying from 97.5 to 101.9%, with standard deviation in the range of 0.6 to 1.9. The results were statistically compared with those of the reference method by applying Student's t-test and F-test. Accuracy evaluated by means of the spike recovery method, range from 97.0 to 106.0%, with precision better than 3%.     

Optimized and validated spectrophotometric methods for the determination of raloxifene in pharmaceuticals using permanganate

Two simple and sensitive spectrophotometric methods are described for the determination of raloxifene hydrochloride (RLX) in pure form and in tablets. The first method (method A) is based on the formation of a yellowish-brown chromogen peaking at 430 nm when RLX was reacted with permanganate in acetic acid medium. In the second method (method B), RLX was reacted with a measured excess of permanganate in H₂SO₄ medium followed by the spectrophotometric measurement of the unreacted KMnO₄ at 550 nm. Under the optimized experimental conditions, Beer’s law is obeyed in the concentration range 0.6–6.0 and 1.5–15.0 μg mL⁻¹ with molar absorptivity of 7.01 × 10⁴ and 2.8 × 10⁴ L mol⁻¹ cm⁻¹ for method A and method B, respectively. The limits of detection (LOD) and quantification (LOQ) have also been reported. The intra-day and inter-day RSD and RE values at three different concentrations were assessed. The proposed methods were applied to the commercially available tablets, and the results were statistically compared with those of the reference method. The accuracy and reliability of the methods were further ascertained by recovery studies.     

Ion-selective sensors for determining ampicillin and oxacillin in biological fluids and pharmaceuticals

Ion-selective sensors based on organic ion exchangers of the ampicillin (oxacillin)-tetradecylammonium (Am-TDA; Ox-TDA) type sensitive to antibiotics of the penicillin group have been developed. The ratio of components (1: 1) and the solubility products of ion associates (8.5 × 10⁻⁸ and 5.2 × 10⁻¹⁰ for Am-TDAand Ox-TDA, respectively) are determined. According to thermoanalytical data, both Am-TDAand Ox-TDAion associates are individual substances not containing water. The organic ion exchangers do not undergo changes on heating up to 70°C. It is established that the electrode functions are linear in the interval from 1 × 10 ⁻⁵ to 1 × 10⁻¹ M for all antibiotics studied. The slopes of the electrode functions for Am and Ox are 58 × 1 and 60 × 1, respectively, being close to the theoretical values for singly charged ions. Methods for the determination of Am and Ox in the blood serum and saliva and in various medicinal forms (tablets, powders in bottles, etc.) are described. __________ Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 40, No. 3, pp. 53–55, March, 2006.     

Tetracycline-selective electrode for content determination and dissolution studies of pharmaceuticals by flow-injection analysis (FIA)

The present work describes the construction and evaluation of different tetracycline (TC)-selective electrodes without inner reference solution and with polymer membranes. The several electrodes were prepared with poly(vinyl chloride) or ethylene(vinyl acetate) membranes comprising o-nitrophenyl octyl ether or bis(2-ethylhexyl)sebacate as mediator solvents and tetracycline tetrakis(4-clorophenyl)borate as ion exchanger. The best performance was recorded for the poly(vinyl chloride) membranes with bis(2-ethylhexyl)sebacate. Using solutions with adjusted ionic strength, this type of electrode presented a slope of 57.4 mV decade(-1) and a reproducibility of +/-0.3 mV day(-1), for an analytical range from 1.2 x 10(-4) to 1.0 x 10(-2) M. The pH working range was 2.0-3.8. Tubular-shaped potentiometric detectors based on the same selective membrane were also constructed. When TC solutions with adjusted ionic strength of concentrations ranging from 1.0 x 10(-4) to 1.0 x 10(-2) M were injected into a single-channel flow manifold, the detectors presented a slope of 56.6 mV decade(-1) and a reproducibility of +/-0.5 mV day(-1). The pH working range was 1.9-3.9. Both batch and flow procedures were applied to the potentiometric analysis of oral dosage forms. Average recoveries were within 98.6 to 100.3% and the t test indicated the accuracy of these results in comparison to an independent methodology. The flow system with the potentiometric detector was employed in dissolution studies as well.     

Recent applications of flow-injection and sequential-injection analysis techniques to chemiluminescence determination of pharmaceuticals

A review is presented on the state of the art of the chemiluminescence analysis of pharmaceuticals by the two most relevant automated controlled-flow methodologies--flow-injection analysis (FIA) and sequential-injection analysis (SIA). The current chemiluminometric applications of FIA and SIA in pharmaceutical analysis are discussed with special emphasis on the analytical figures of merit and sample matrix characteristics. The review involving 211 references and covering papers published between 2001 and 2006 is divided into several sections according to the fundamental types of chemiluminescence systems employed.     

Spectrophotometric studies and application of imipramine-eriochrome cyanine R system for determination of imipramine in pharmaceuticals

Eriochrome cyanine R (ECR) has been tested as reagent for the determination of imipramine. It reacts in neutral medium with imipramine forming reddish compound, which can be quantitatively extracted into n-butanol. This property has been successfully used for the extractive-spectrophotometric determination of imipramine. Beer's law is obeyed in concentration range of 10-80 microg ml(-1) of imipramine. The method was applied to the determination of imipramine in its pharmaceutical.     

Rapid methods for the microbiological surveillance of pharmaceuticals

The use of rapid microbiological methods in pharmaceutical laboratories has improved the quality control analysis of water, products, raw materials, and enhanced the antimicrobial effectiveness testing of pharmaceutical finished products. Rapid release of samples has resulted in the optimization of manufacturing, product testing, and release allowing high throughput and simultaneous analysis of pharmaceutical formulations. ATP Bioluminescence, Impedance, Direct Viable Counts, and Flow Cytometry determine the total microbial content in a given pharmaceutical sample while PCR and Immunoassays detect the presence or absence of specific microbial species. Rapid methods provide reliable and cost effective analysis for the microbiological evaluation of pharmaceutical environments. The dramatic reduction in detection times and analysis, e.g., from 30 hours to 90 minutes, by using rapid methods will ultimately lead the pharmaceutical industry closer to real time monitoring of processes and samples.     

Spectrophotometric determination of hydralazine with 2-hydroxy-1-naphthaldehyde in pharmaceuticals

A new extraction-spectrophotometric method for the determination of hydralazine, based on its reaction with 2-hydroxy-1-naphthaldehyde at 25 degrees C, is described. The calibration curve was linear between 0.4 and 6 mg/mL of hydralazine. The molar absorbtivity of the product at 408 nm is 40,900 L.mol-1.cm-1. The method described was applied to the analysis of hydralazine in pharmaceutical preparations containing reserpine, hydrochlorothiazide, bendrofluorthiazine, propranolol, and other substances. The agreement with the U.S.P. XXI method was satisfactory for tablets and injections, but not for pellets.     

Voltammetric and flow amperometric methods for the determination of melatonin in pharmaceuticals

Melatonin can be sensitively detected in pharmaceuticals by two different and simple electrochemical methods: cyclic voltammetry (CV) and amperometric detection in a flow injection analysis system (FIA-ED). An adequate pre-treatment of the carbon paste electrode in the first case and the employ of a high flow rate in the second one were the key for obtaining a very good reproducibility (R.S.D. values of 1.5 (n=10) and 1.3% (n=20), respectively). Low limits of detection were achieved and with the coupling of a flow system a linear dynamic range of three orders of magnitude (from 10(-8) to 10(-5) M) was obtained. Both methods were applied to the determination of melatonin in pharmaceuticals. In order to best validate these methodologies a fluorescent procedure was developed to contrast the results. As no interferences from the matrix were found the employ of a separation technique is not necessary. In this way the procedure is fastened and simplified. Moreover, the low price, ease of handling, possibility of automation and high sample throughput are important advantages that convert the flow methodology in an attractive alternative for quality control of pharmaceuticals.     

Spectrophotometric methods for the determination of sulphathiourea

Three simple and sensitive spectrophotometric methods are described for the determination of sulphathiourea. The methods are based on the interaction of this thioamide with either iodine, tetracyanoethylene (TCNE) or copper nitrate to give the corresponding charge-transfer or metal-ion complexes, which can be measured at 363, 350 and 353 nm, respectively. In each case, a 1:1 complex was formed. Beer's law is obeyed for each procedure in a concentration range of 1-150 micrograms ml-1. The proposed procedures can be applied to the determination of sulphathiourea in its pharmaceutical formulations.     

NMR methods for determining disulfide-bond connectivities

Animal toxins are the major class of secreted disulfide-rich proteins, with ∼70% containing two or more disulfide bonds. Incorrect pairing of these disulfide bonds typically leads to a non-native three-dimensional fold accompanied by a loss of protein function. Determination of the native disulfide-bond framework is therefore a key component in the structural characterization of toxins. In this article, we review NMR approaches for elucidation of disulfide-bond connectivities. A major advantage of these NMR approaches is that they are non-invasive, leaving the sample intact at the end of the analysis for use in other studies.     

Phenotypic screening for pharmaceuticals using tissue constructs

Compounds can be screened for pharmaceutical activity either by detecting interactions with specified target molecules such as receptors or enzymes (molecular screening) or observing effects on the structure or physiological activities of cells or tissues (phenotypic screening). Screening at the molecular level has been greatly enhanced by fluorescence methods. Especially the combination of confocal detection with measurements of the amplitudes and time courses of fluorescence fluctuations have reduced sample volumes to < microliters and have increased throughputs to >100000 compounds per day. Screening at the molecular level, however, does not provide information about the effects of test compounds on cellular functions. Phenotypic screening, although much slower than molecular screening, does provide information about effects on cell or tissue structure or function and therefore can be used to eliminate at an early stage compounds that are toxic or do not produce the desired cellular response. Tissue constructs reconstituted using cells of specified types and defined extracellular matrix components provide test systems for detecting the effects of test compounds on cellular mechanical functions such as the development of contractile force and on cell and matrix structure and stiffness. For example, constructs based on vascular smooth muscle cells provide information about effects on cellular contractile force that can be used to identify agents that control blood pressure. Tissue constructs that mimic skeletal, smooth and heart muscles and connective tissues have been produced and can be used to study mechanical and structural responses to active compounds.     

New spectrophotometric methods for the quantitative determination of chloramphenicol in pharmaceuticals

Three new methods are proposed for the simple and sensitive spectrophotometric determination of chloramphenicol in pure form and in related pharmaceutical preparations. Two methods are based on the diazotization of reduced chloramphenicol, followed by coupling either with iminodibenzyl to give a violet colored product (λ_max = 590 nm) or with 3-aminophenol to produce an orange-red colored product (λ_max = 470 nm). The third method makes use of sodium molybdate and pyrocatechol as coupling agents to give a pale red colored product (λ_max = 490 nm). The proposed procedures are highly reproducible and the maximum relative standard deviation is 0.3 % for five determinations. The new methods have been applied to the determination of chloramphenicol in capsules and injection solutions, and the results compare favorably with the official method. Common auxiliary substances used as components of pharmaceutical preparations do not interfere with the proposed methods. Plausible reaction mechanisms are proposed. Published in Khimiko-Farmatsevticheskii Zhurnal, Vol. 40, No. 10, pp. 52–56, October, 2006.     

A rationale for determining, testing, and controlling specific impurities in pharmaceuticals that possess potential for genotoxicity

The synthesis of pharmaceutical products frequently involves the use of reactive reagents and the formation of intermediates and by-products. Low levels of some of these may be present in the final drug substance and drug product as impurities. Such chemically reactive impurities may have at the same time the potential for unwanted toxicities including genotoxicity and carcinogenicity and hence can have an impact on product risk assessment. This paper outlines a procedure for testing, classification, qualification, toxicological risk assessment, and control of impurities possessing genotoxic potential in pharmaceutical products. Referencing accepted principles of cancer risk assessment, this document proposes a staged threshold of toxicological concern (TTC) approach for the intake of genotoxic impurities over various periods of exposure. This staged TTC is based on knowledge about tumorigenic potency of a wide range of genotoxic carcinogens and can be used for genotoxic compounds, for which cancer data are limited or not available. The delineated acceptable daily intake values of between approximately 1.5 microg/day for approximately lifetime intake and approximately 120 microg/day for < or = 1 month are virtually safe doses. Based on sound scientific reasoning, these virtually safe intake values do not pose an unacceptable risk to either human volunteers or patients at any stage of clinical development and marketing of a pharmaceutical product. The intake levels are estimated to give an excess cancer risk of 1 in 100,000 to 1 in a million over a lifetime, and are extremely conservative given the current lifetime cancer risk in the population of over 1 in 4 (http://seer.cancer.gov/statfacts/html.all.html). The proposals in this document apply to all clinical routes of administration and to compounds at all stages of clinical development. It is important to note that certain types of products, such as those for life-threatening indications for which there are no safer alternatives, allow for special considerations using adaptations of the principles outlined in this paper.