Sevgi Tatar Ulu

Spectrofluorimetric determination of fluoroquinolones in pharmaceutical preparations

Simple, rapid and highly sensitive spectrofluorimetric method is presented for the determination of four fluoroquinolone (FQ) drugs, ciprofloxacin, enoxacin, norfloxacin and moxifloxacin in pharmaceutical preparations. Proposed method is based on the derivatization of FQ with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in borate buffer of pH 9.0 to yield a yellow product. The optimum experimental conditions have been studied carefully. Beers law is obeyed over the concentration range of 23.5-500 ng mL(-1) for ciprofloxacin, 28.5-700 ng mL(-1) for enoxacin, 29.5-800 ng mL(-1) for norfloxacin and 33.5-1000 ng mL(-1) for moxifloxacin using NBD-Cl reagent, respectively. The detection limits were found to be 7.0 ng mL(-1) for ciprofloxacin, 8.5 ng mL(-1) for enoxacin, 9.2 ng mL(-1) for norfloxacin and 9.98 ng mL(-1) for moxifloxacin, respectively. Intra-day and inter-day relative standard deviation and relative mean error values at three different concentrations were determined. The low relative standard deviation values indicate good precision and high recovery values indicate accuracy of the proposed methods. The method is highly sensitive and specific. The results obtained are in good agreement with those obtained by the official and reference method. The results presented in this report show that the applied spectrofluorimetric method is acceptable for the determination of the four FQ in the pharmaceutical preparations. Common excipients used as additives in pharmaceutical preparations do not interfere with the proposed method.

Rapid and sensitive spectrofluorimetric determination of enrofloxacin, levofloxacin and ofloxacin with 2,3,5,6-tetrachloro-p-benzoquinone.

A highly sensitive spectrofluorimetric method was developed for the first time, for the analysis of three fluoroquinolones (FQ) antibacterials, namely enrofloxacin (ENR), levofloxacin (LEV) and ofloxacin (OFL) in pharmaceutical preparations through charge transfer (CT) complex formation with 2,3,5,6-tetrachloro-p-benzoquinone (chloranil,CLA). At the optimum reaction conditions, the FQ-CLA complexes showed excitation maxima ranging from 359 to 363nm and emission maxima ranging from 442 to 488nm. Rectilinear calibration graphs were obtained in the concentration range of 50-1000, 50-1000 and 25-500ngmL(-1) for ENR, LEV and OFL, respectively. The detection limit was found to be 17ngmL(-1) for ENR, 17ngmL(-1) for LEV, 8ngmL(-1) for OFL, respectively. Excipients used as additive in commercial formulations did not interfere in the analysis. The method was validated according to the ICH guidelines with respect to specificity, linearity, accuracy, precision and robustness. The proposed method was successfully applied to the analysis of pharmaceutical preparations. The results obtained were in good agreement with those obtained using the official method; no significant difference in the accuracy and precision as revealed by the accepted values of t- and F-tests, respectively.

Highly sensitive spectrofluorimetric determination of lomefloxacin in spiked human plasma, urine and pharmaceutical preparations

A sensitive, simple and selective spectrofluorimetric method was developed for the determination of lomefloxacin in biological fluids and pharmaceutical preparations. The method is based on the reaction between the drug and 4-chloro-7-nitrobenzodioxazole in borate buffer of pH 8.5 to yield a highly fluorescent derivative that is measured at 533 nm after excitation at 433 nm. The calibration curves were linear over the concentration ranges of 12.5-625, 15-1500 and 20-2000 ng/mL for plasma, urine and standard solution, respectively. The limits of detection were 4.0 ng/mL in plasma, 5.0 ng/mL in urine and 7.0 ng/mL in standard solution. The intra-assay accuracy and precision in plasma ranged from 0.032 to 2.40% and 0.23 to 0.36%, respectively, while inter-assay accuracy and precision ranged from 0.45 to 2.10% and 0.25 to 0.38%, respectively. The intra-assay accuracy and precision estimated on spiked samples in urine ranged from 1.27 to 4.20% and 0.12 to 0.24%, respectively, while inter-assay accuracy and precision ranged from 1.60 to 4.00% and 0.14 to 0.25%, respectively. The mean recovery of lomefloxacin from plasma and urine was 98.34 and 98.43%, respectively. The method was successfully applied to the determination of lomefloxacin in pharmaceuticals and biological fluids.

Development of an HPLC method for the determination of mexiletine in human plasma and urine by solid-phase extraction.

A sensitive and specific high-performance liquid chromatography (HPLC) method has been developed and validated for the quantification of mexiletine (MEX) in human plasma and urine. It uses solid-phase extraction (SPE) followed by an automated reversed-phase HPLC with a pre-column derivatization with 4-chloro-7-nitrobenzofurazan (NBD-CI) and UV-vis Absorbance detection. The process was set as: the UV-vis Absorbance wavelength was set at 458nm. Chromatographic separation was performed on a Phenomenex-C(18) Column (Aqua, 150mmx4.6mm i.d. with 5mum particle size) with the mobile phase consisting of acetonitrile and water (80:20, v/v), and the flow rate was set at 1.0mLmin(-1). Calibration of the overall analytical procedure gave a linear signal (r>0.9998) over a MEX concentration range of 0.2-2.0mugmL(-1) in human plasma and urine. The detection limit in plasma and urine was 0.1mugmL(-1). Intra- and inter-day precision of the assay at three concentrations within this range were 0.31-2.50%. The high specificity and sensitivity have been achieved by this fast method (total run-time <6min). The method has been successfully validated in human plasma and urine and it has been shown to be precise, accurate and reliable.

Spectrophotometric method for the determination, validation, spectroscopic and thermal analysis of diphenhydramine in pharmaceutical preparation

A sensitive, simple and rapid spectrophotometric method was developed for the determination of diphenhydramine in pharmaceutical preparation. The method was based on the charge-transfer complex of the drug, as n-electron donor, with 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ), as pi-acceptor. The formation of this complex was also confirmed by UV-vis, FTIR and (1)H NMR spectra techniques and thermal analysis. The proposed method was validated according to the ICH guidelines with respect to linearity, limit of detection, limit of quantification, accuracy, precision, recovery and robustness. The linearity range for concentrations of diphenhydramine was found to be 12.5-150 microg/mL with acceptable correlation coefficients. The detection and quantification limits were found to be 2.09 and 6.27 microg/mL, respectively. The proposed and references methods were applied to the determination of drug in syrup. This preparation were also analyzed with an reference method and statistical comparison by t- and F-tests revealed that there was no significant difference between the results of the two methods with respect to mean values and standard deviations at the 95% confidence level.

UV–Second Derivative Spectrophotometric and Colorimetric Methods for the Determination, Validation and Thermogravimetric Analysis of New Oral Antidiabetic Pioglitazone in Pure and Pharmaceutical Preparations

Abstract Three rapid, sensitive, and simple spectrophotometric methods have been developed for the determination of pioglitazone in pure and pharmaceutical preparations. For the first method, UV-spectrophotometry, standard solutions were measured at 270.2 nm. The first method was linear from 5.0–20.0 µgmL−1. The linearity was found to be 5.0–20.0 µgmL−1. For the second method, the distances between two extremum values (peak-to-peak amplitudes), 272.0 and 287.4 nm were measured in the second order derivative-spectra of standard solutions. Calibration curves were constructed by plotting d2 A/dλ2 values against concentrations, 2.0–12.0 µgmL−1 of pioglitazone standards in acetonitrile. The detection limits of pioglitazone were 0.10 and 0.16 µgmL−1 for UV and derivative spectrophotometric methods, respectively. The third method was based on the formation of an ion association complex with bromocresol green (BCG), bromocresol purple (BCP), bromophenol blue (BPB), and bromothymol blue (BTB). The assay was linear over the concentration range of 20.0–100.0 µgmL−1 for BCG, 10.0–100.0 µgmL−1 for BCP, 20.0–120.0 µgmL−1 for BPB, and 10.0–100.0 µgmL−1 for BTB. The detection limits of pioglitazone was found to be 0.14 µg mL−1 for BCG, 0.32 for BCP, 1.24 µgmL−1 for BPB, and 0.22 µgmL−1 for BTB. The thermal analysis of the pioglitazone was studied by Thermogravimetric Analysis-Differential Scanning Calorimetry (TGA-DSC) techniques. Enthalpy change of pioglitazone was found to be 85.16 J/g. The proposed methods were validated according to the ICH guidelines (1996) with respect to specificity, linearity, limits of detection and quantification, accuracy, precision, and robustness. The results demonstrated that the procedure is accurate, precise, specific, and reproducible (percent relative standard deviation <2%), while being simple and less time consuming. The three proposed methods have been successfully applied to the assay of pioglitazone in pure and in pharmaceutical preparations. The results compared with those obtained by an ultraviolet spectrophotometric method using t and F tests.

An HPLC method for the determination of bisoprolol in human plasma and its application to a pharmacokinetic study.

A new high-performance liquid chromatographic method is described for the determination of bisoprolol in human plasma. The proposed method was based on the derivatization of bisoprolol with 4-chloro-7-nitro-2,1,3-benzoxadiazole in borate buffer at pH 9.5 to yield a fluorescent product. Chromatographic separation of bisoprolol was achieved by using isocratic elution at a flow rate of 1.2 mL/min on a C18 reversed-phase column (Inertsil, 4 μm, 150 4.6 mm) at 40°C. The mobile phase used for the analysis was methanol-water (70:30, % v/v). Fluorescence detector was used at the excitation and emission wavelengths of 458 and 525 nm, respectively. The method was validated for linearity, limit of detection, limit of quantification, precision, accuracy, recovery and system suitability. The assay was linear over the concentration range of 10-2000 ng/mL. This method was applied in pharmacokinetic studies of bisoprolol preparations in healthy volunteers.

Sensitive spectrofluorimetric determination of tizanidine in pharmaceutical preparations, human plasma and urine through derivatization with dansyl chloride.

A sensitive spectrofluorimetric method was developed for the determination of tizanidine in human plasma, urine and pharmaceutical preparations. The method is based on reaction of tizanidine with 1-dimethylaminonaphthalene-5-sulphonyl chloride (dansyl chloride) in an alkaline medium to form a highly fluorescent derivative that was measured at 511 nm after excitation at 383 nm. The different experimental parameters affecting the fluorescence intensity of tizanidine was carefully studied and optimized. The fluorescence-concentration plots were rectilinear over the ranges 50-500 and 20-300 ng/mL for plasma and urine, respectively, detection limits of 1.81 and 0.54 ng/mL and quantification limits of 5.43 and 1.62 ng/mL for plasma and urine, respectively. The method presents good performance in terms of linearity, detection and quantification limits, precision, accuracy and specificity. The proposed method was successfully applied for the determination of tizanidine in pharmaceutical preparations. The results obtained were compared with a reference method, using t- and F-tests.

Determination of Bupropion Using Liquid Chromatography with Fluorescence Detection in Pharmaceutical Preparations, Human Plasma and Human Urine

A novel pre-column derivatization reversed-phase high-performance liquid chromatography with fluorescence detection is described for the determination of bupropion in pharmaceutical preparation, human plasma and human urine using mexiletine as internal standard. The proposed method is based on the reaction of 4-chloro-7-nitrobenzofurazan (NBD-Cl) with bupropion to produce a fluorescent derivative. The derivative formed is monitored on a C18 (150 mm × 4.6 mm i.d., 5 µm) column using a mobile phase consisting of methanol-water 75:25 (v/v), at a flow-rate of 1.2 mL/min and detected fluorimetrically at λ(ex) = 458 and λ(em) = 533 nm. The assay was linear over the concentration ranges of 5-500 and 10-500 ng/mL for plasma and urine, respectively. The limits of detection and quantification were calculated to be 0.24 and 0.72 ng/mL for plasma and urine, respectively (inter-day results). The recoveries obtained for plasma and urine were 97.12% ± 0.45 and 96.00% ± 0.45, respectively. The method presents good performance in terms of precision, accuracy, specificity, linearity, detection and quantification limits and robustness. The proposed method is applied to determine bupropion in commercially available tablets. The results were compared with an ultraviolet spectrophotometry method using t- and F-tests.

Validated spectrofluorimetric method for the determination of tamsulosin in spiked human urine, pure and pharmaceutical preparations

A novel, sensitive and selective spectrofluorimetric method was developed for the determination of tamsulosin in spiked human urine and pharmaceutical preparations. The proposed method is based on the reaction of tamsulosin with 1-dimethylaminonaphthalene-5-sulfonyl chloride in carbonate buffer pH 10.5 to yield a highly fluorescent derivative. The described method was validated and the analytical parameters of linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, precision, recovery and robustness were evaluated. The proposed method showed a linear dependence of the fluorescence intensity on drug concentration over the range 1.22 × 10(-7) to 7.35 × 10(-6)  M. LOD and LOQ were calculated as 1.07 × 10(-7) and 3.23 × 10(-7)  M, respectively. The proposed method was successfully applied for the determination of tamsulosin in pharmaceutical preparations and the obtained results were in good agreement with those obtained using the reference method.