Safaa M. Riad

Multi-residues determination of antimicrobials in fish tissues by HPLC–ESI-MS/MS method

A rapid, simple, sensitive and specific LC-MS/MS method was developed and validated for the simultaneous quantification of four antimicrobials commonly used in aquaculture, namely ciprofloxacin (CPX), trimethoprim (TMP), sulphadimethoxine (SDM) and florphenicol (FLOR) in fish tissues. The LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. Sample preparation involves simple liquid extraction step followed by post-extraction clean-up step with n-hexane. The purified extracts were chromatographed on Agilent Poroshell 120 EC, C18 (50 mm × 3 mm, 2.7 μm) column by pumping an isocratic mobile phase consisting of 0.1% formic acid in water:0.1% formic acid in methanol (20:80, by volume) at a flow rate of 0.4 mL/min. A detailed validation of the method was performed as per FDA guidelines and the standard curves were found to be linear in the range of 1-100 ng/g for both CPX and TMP, 0.5-100 ng/g for SDM and 1-50 ng/g for FLOR. The intra-day and inter-day precision and accuracy of the results were within the acceptable limits. A run time of 1.5 min for each sample made it possible to analyze multiple fish tissue samples per day. The developed assay method was successfully applied for the detection of antimicrobials in real fish tissue samples obtained from different fish farms.

Novel poly (vinyl chloride) matrix membrane electrodes for the determination of phenolic pollutants in waste water

The construction and electrochemical response characteristics of poly (vinyl chloride) matrix membrane sensors for seven phenolic compounds (phenol, o-cresol, p-cresol, p-chlorophenol, o-nitrophenol, alpha-naphthol and beta-naphthol) are described. The membranes incorporate ion association complexes of these seven phenolic anions with phenanthroline-iron(II) as electroactive materials. These sensors show linear response for phenol, o-cresol, p-cresol, p-chlorophenol, o-nitrophenol, alpha-naphthol, and beta-naphthol over wide concentration ranges, with an average anionic slope 54.3 mV per concentration decade. The suggested sensors exhibit fast response time (1 min), low determination limits (1 x 10(-5)M), good stability (2-3 weeks), reasonable selectivity to phenolic compounds in the presence of other water pollutants. The average percentage recovery was 99.78+/-0.088 for individual phenolates and 99.61+/-0.198 for phenolates in mixtures. The investigated sensors were successfully used for direct potentiometric determination of traces of these phenolic compounds in wastewater samples. Results with mean accuracy of 99.74+/-0.29%, 99.82+/-0.36%, 99.65+/-0.47%, 99.73+/-0.37%, 99.77+/-0.30%, 99.86+/-0.31% and 99.91+/-0.22% was obtained for the seven sensors, respectively. These results were compared with data obtained using the British pharamacopial method (The British Pharmacopoeia, Her Majestys Stationary Office, London, 1993) and others (Clarkes Isolation and Identification of Drugs in Pharmaceuticals Body Fluids and Post-mortem Materials, 2nd Edition, The Pharmaceutical Press, London, 1998).

Stability indicating spectrophotometric and spectrodensitometric methods for the determination of diatrizoate sodium in presence of its degradation product.

Three sensitive, selective, and precise stability indicating methods for the determination of the X-ray contrast agent, diatrizoate sodium (DTA), in the presence of its acidic degradation product (highly cytotoxic 3,5 diamino metabolite) and in pharmaceutical formulation were developed and validated. The first method is a first derivative (D1) spectrophotometric one, which allows the determination of DTA in the presence of its degradate at 231.2 nm (corresponding to zero crossing of the degradate) over a concentration range of 2-24 μg/mL with mean percentage recovery 99.95±0.97%. The second method is the first derivative of the ratio spectra (DD1) by measuring the peak amplitude at 227 nm over the same concentration range as D1 spectrophotometric method, with mean percentage recovery 99.99±1.15%. The third method is a TLC-densitometric one, where DTA was separated from its degradate on silica gel plates using chloroform:methanol:ammonium hydroxide (20:10:2 by volume) as a developing system. This method depends on quantitative densitometric evaluation of thin layer chromatogram of DTA at 238 nm over a concentration range of 4-20 μg/spot, with mean percentage recovery 99.88±0.89%. The selectivity of the proposed methods was tested using laboratory-prepared mixtures. The proposed methods have been successfully applied to the analysis of DTA in pharmaceutical dosage forms without interference from other dosage form additives. The results were statistically compared with the official US pharmacopeial method. No significant difference for either accuracy or precision was observed.

Simultaneous Determination of Metronidazole and Diiodohydroxyquine in Bulk Powder and Paramibe Compound Tablets by TLC-Densitometry and HPLC

Two sensitive and precise chromatographic methods were developed and validated for simultaneous determination of metronidazole (MTR) and diiodohydroxyquin (DIQ) in pharmaceutical preparation. The techniques adopted for quantification are coupled TLC-densitometry and HPLC. A mixture of chloroform, toluene, ethanol and acetic acid (9:9:1:1, v/v/v/v) was used as the developing solvent for TLC-densitometry. A mixture of methanol and acetonitrile, (96:4, v/v) was used as a mobile phase for HPLC at 0.6 mL min-1 flow rate and UV detection at 254 nm. Linearity was obtained in concentration range of 0.5-10 μg spot-1 for DIQ and 1-20 μg spot-1 for MTR applying TLC-densitometry and 0.005-0.5 mg mL-1 for DIQ and 0.01-0.5 mg mL-1 for MTR applying HPLC. The selectivity of the proposed methods was checked using laboratory prepared mixtures. The proposed methods were successfully applied to the analysis of MTR and DIQ in their mixture and in pharmaceutical dosage forms without interference from other additives.

A comparative study between three stability indicating spectrophotometric methods for the determination of diatrizoate sodium in presence of its cytotoxic degradation product based on two-wavelength selection

Three sensitive, selective, and precise stability indicating spectrophotometric methods for the determination of the X-ray contrast agent, diatrizoate sodium (DTA) in the presence of its acidic degradation product (highly cytotoxic 3,5-diamino metabolite) and in pharmaceutical formulation, were developed and validated. The first method is ratio difference, the second one is the bivariate method, and the third one is the dual wavelength method. The calibration curves for the three proposed methods are linear over a concentration range of 2-24 μg/mL. The selectivity of the proposed methods was tested using laboratory prepared mixtures. The proposed methods have been successfully applied to the analysis of DTA in pharmaceutical dosage forms without interference from other dosage form additives. The results were statistically compared with the official US pharmacopeial method. No significant difference for either accuracy or precision was observed.

A New Platform for Profiling Degradation-Related Impurities Via Exploiting the Opportunities Offered by Ion-Selective Electrodes: Determination of Both Diatrizoate Sodium and Its Cytotoxic Degradation Product

Although the ultimate goal of administering active pharmaceutical ingredients (APIs) is to save countless lives, the presence of impurities and/or degradation products in APIs or formulations may cause harmful physiological effects. Today, impurity profiling (i.e., the identity as well as the quantity of impurity in a pharmaceutical) is receiving critical attention from regulatory authorities. Despite the predominant use of spectroscopic and chromatographic methods over electrochemical methods for impurity profiling of APIs, this work investigates the opportunities offered by electroanalytical methods, particularly, ion-selective electrodes (ISEs), for profiling degradation-related impurities (DRIs) compared with conventional spectroscopic and chromatographic methods. For a meaningful comparison, diatrizoate sodium (DTA) was chosen as the anionic X-ray contrast agent based on its susceptibility to deacetylation into its cytotoxic and mutagenic degradation product, 3,5-diamino-2,4,6 triiodobenzoic acid (DTB). This cationic diamino compound can be also detected as an impurity in the final product because it is used as a synthetic precursor for the synthesis of DTA. In this study, four novel sensitive and selective sensors for the determination of both DTA and its cytotoxic degradation products are presented. Sensors I and II were developed for the determination of the anionic drug, DTA, and sensors III and IV were developed for the determination of the cationic cytotoxic impurity. The use of these novel sensors not only provides a stability-indicating method for the selective determination of DTA in the presence of its degradation product, but also permits DRI profiling. Moreover, a great advantage of these proposed ISE systems is their higher sensitivity for the quantification of DTB relative to other spectroscopic and chromatographic methods, so it can measure trace amounts of DTB impurities in DTA bulk powder and pharmaceutical formulation without a need for preliminary separation.

Validated univariate and multivariate spectrophotometric methods for the determination of pharmaceuticals mixture in complex wastewater.

Five, accurate, precise, and sensitive univariate and multivariate spectrophotometric methods were developed for the simultaneous determination of a ternary mixture containing Trimethoprim (TMP), Sulphamethoxazole (SMZ) and Oxytetracycline (OTC) in waste water samples collected from different cites either production wastewater or livestock wastewater after their solid phase extraction using OASIS HLB cartridges. In univariate methods OTC was determined at its λmax 355.7 nm (0D), while (TMP) and (SMZ) were determined by three different univariate methods. Method (A) is based on successive spectrophotometric resolution technique (SSRT). The technique starts with the ratio subtraction method followed by ratio difference method for determination of TMP and SMZ. Method (B) is successive derivative ratio technique (SDR). Method (C) is mean centering of the ratio spectra (MCR). The developed multivariate methods are principle component regression (PCR) and partial least squares (PLS). The specificity of the developed methods is investigated by analyzing laboratory prepared mixtures containing different ratios of the three drugs. The obtained results are statistically compared with those obtained by the official methods, showing no significant difference with respect to accuracy and precision at p=0.05.

Qualitative and Quantitative Chemometry as Stability-Indicating Methods for Determination of Dantrolene Sodium and Paracetamol

Methods and Result: Three stability indicating chemometric methods are applied for the simultaneous determination of Dantrolene sodium (DNT) and Paracetamol (PAR). Partial Least Squares (PLS), Concentration Residuals Augmented Classical Least Squares (CRACLS) and Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) were selected for that purpose. DNT and PAR were determined in the linearity range of (2 – 10 g mL) and (12 – 28 g mL), respectively, in the presence of their degradation products. The presented methods were compared for their qualitative and quantitative analyses and validated according to the ICH guidelines. Furthermore, statistical comparison between the results obtained by the proposed methods and the reported chromatographic method showed no significant differences.

Three Different Spectrophotometric Methods Exploiting Ratio Spectra for the Selective Determination of Iohexol in the Presence of its Acidic Degradate

Conclusion: Iohexol has been successfully determined in its pure form and pharmaceutical formulation (Omnipaque ® vials) utilizing the proposed methods with no interference from the present additives. The results obtained by each of the proposed methods were statistically compared to the official United States pharmacopeial method and non-significant difference was obtained regarding accuracy or precision.

Monitoring of the degradation kinetics of diatrizoate sodium to its cytotoxic degradant using a stability-indicating high-performance liquid chromatographic method.

The X-ray diagnostic agent sodium diatrizoate (DTA) was studied for chemical degradation. The 3,5-diamino derivative was found to be the alkaline and acidic degradation product. The 3,5-diamino degradate is also the synthetic precursor of DTA and it is proved to have cytotoxic and mutagenic effects. A sensitive, selective and precise high-performance liquid chromatographic stability-indicating method for the determination of DTA in the presence of its acidic degradation product and in pharmaceutical formulation was developed and validated. Owing to the high toxicity of the degradation product, the kinetics of the acidic degradation process was monitored by the developed RP-HPLC method. The reaction was found to follow pseudo-first order kinetics. The kinetic parameters such as rate constant (K) and half-life (t½ ) were calculated under different temperatures and acid concentrations; activation energy was estimated from the Arrhenius plot. The developed RP-HPLC method depends on isocratic elution of a mobile phase composed of methanol-water (25:75 v/v; pH adjusted with phosphoric acid), and UV detection at 238 nm. The method showed good linearity over a concentration range of 2-100 μg/mL with mean percentage recovery of 100.04 ± 1.07. The selectivity of the proposed method was tested using laboratory-prepared mixtures. The proposed method has been successfully applied to the analysis of DTA in pharmaceutical dosage forms without interference from other dosage form additives and the results were statistically compared with the official USP method. Validation of the proposed method was performed according to International Conference on Harmonization guidelines.