Sequential SERS determination of aspirin and vitamin C using in situ laser-induced photochemical silver substrate synthesis in a moving flow cell.

In this contribution, the utility of sequential injection analysis in combination with surface-enhanced Raman spectroscopy (SERS) as a detection technique was investigated for simultaneous determination of aspirin and vitamin C in their pharmaceutical dosage forms and in spiked urine samples. The silver substrate was synthesized in situ by laser-induced photochemical procedure. By focusing the laser on a flow cell at 1 ml/min of continuous flow of 0.5 mM silver nitrate and 5 mM sodium citrate mixture, an active silver spot on the inner wall of the flow cell was prepared in a few seconds. The whole setup is fully computer controlled using ATLAS software to combine the two techniques. The system allows sequential determination of aspirin concentrations ranging from 100 to 500 ng/ml and vitamin C concentrations between 10 and 110 ng/ml with good precision of relative standard deviations (RSDs) of 0.85 and 1.7 %, respectively. A comparison of these results with those of the reported procedures showed excellent results compared with t and F values, indicating good accuracy and precision. The detection limits were 32 and 3 ng/ml for aspirin and vitamin C, respectively.

Utility of surface enhanced Raman spectroscopy (SERS) for elucidation and simultaneous determination of some penicillins and penicilloic acid using hydroxylamine silver nanoparticles.

Elucidation and quantitative determination of some of commonly used penicillins (ampicillin, penicillin G and carbenicillin) in the presence of their main degradation product (penicilloic acid) were developed. Forced acidic and basic degradation processes were applied at different time intervals. The formed degradation products were elucidated and quantified using surface enhanced Raman spectroscopy (SERS). Silver nanoparticles (AgNPs) prepared by reduction of silver nitrate using hydroxylamine-HCl in alkaline medium were used as SERS substrate. The results obtained in SERS were confirmed by the application of LC/MS method. The concentration range was 100-600 ng/ml in case of the studied penicillins and 100-700 ng/ml in case of penicilloic acid. An excellent correlation coefficient was found in case of ampicillin (r=0.9993) and in the case of penicilloic acid (r=0.9997). Validation procedures were carried out including precision, robustness and accuracy by comparing F- and t-values of both the proposed and reported methods.

Antibacterial effect of various shapes of silver nanoparticles monitored by SERS.

A comparative evaluation of antimicrobial effect of synthesized silver nanoparticles (AgNPs) of different shapes using different methods was performed. Spherical, triangular and hexagonal AgNPs with an average size of 40 nm were chemically prepared and characterized by transmission electron microscope (TEM) and UV-visible spectroscopy. The antimicrobial effect of these different AgNPs against the gram negative bacterium Escherichia coli (E. coli) was studied by surface enhanced Raman spectroscopy (SERS), the evaluation of growth curves and inhibition zones. SERS proved to be sensitive to monitor the changes that occurred in the bacterial cells upon interaction with AgNPs, which qualitatively compared well with the data provided by the reference methods. However, as SERS is already sensitive to initial changes in the chemistry of bacteria due to the antibacterial effect of the AgNPs, fast and detailed information is provided by SERS as opposed to the classical reference methods based on the evaluation of growth curves and inhibition zones. The results of this work also demonstrate that hexagonal AgNPs display the highest antibacterial effect when compared to other NPs shapes, with triangular AgNPs exhibiting no antibacterial effect under the adopted conditions.

Highly reproducible SERS detection in sequential injection analysis: real time preparation and application of photo-reduced silver substrate in a moving flow-cell.

This paper reports an improved way for performing highly reproducible surface enhanced Raman scattering of different analytes using an automated flow system. The method uses a confocal Raman microscope to prepare SERS active silver spots on the window of a flow cell by photo-reduction of silver nitrate in the presence of citrate. Placement of the flow cell on an automated x and y stages of the Raman microscope allows to prepare a fresh spot for every new measurement. This procedure thus efficiently avoids any carry over effects which might result from adsorption of the analyte on the SERS active material and enables highly reproducible SERS measurements. For reproducible liquid handling the used sequential injection analysis system as well as the Raman microscope was operated by the flexible LabVIEW based software ATLAS developed in our group. Quantitative aspects were investigated using Cu(PAR)2 as a model analyte. Concentration down to 5×10(-6) M provided clear SERS spectra, a linear concentration dependence of the SERS intensities at 1333 cm(-1) was obtained from 5×10(-5) to 1×10(-3) with a correlation coefficient r=0.999. The coefficient of variation of the method Vxo was found to be 5.6% and the calculated limit of detection 1.7×10(-5) M. The results demonstrate the potential of SERS spectroscopy to be used as a molecular specific detector in aqueous flow systems.

A selective spectrophotometric method for determination of rosoxacin antibiotic using sodium nitroprusside as a chromogenic reagent.

A selective spectrophotometric method for the determination of rosoxacin (ROS), a 4-quinolone antimicrobial agent, has been developed and validated. The method was based on the reaction of ROS with alkaline sodium nitroprusside (SNP) reagent at room temperature forming a red colored chromogen measured at 455 nm. The conditions affecting the reaction (SNP concentration, pH, color-developing time, temperature, diluting solvent and chromogen stability time) were optimized. Under the optimum conditions, good linear relationship (r=0.9987) was obtained between the absorbance and the concentration of ROS in the range of 20-50 microg ml(-1). The assay limits of detection and quantitation were 2.5 and 8.4 microg ml(-1), respectively. The method was successfully applied to the analysis of bulk drug and laboratory-prepared tablets; the mean percentage recoveries were 100.1+/-0.33 and 101.24+/-1.28%, respectively. The results were compared favourably with those obtained by the reported method; no significant difference in the accuracy and precision as revealed by the accepted values of t- and F-tests, respectively. The robustness and ruggedness of the method was checked and satisfactory results were obtained. The proposed method was found to be highly selective for ROS among the fluoroquinolone antibiotics. The reaction mechanism was proposed and it proceeded in two steps; the formation of nitroferrocyanide by the action of sodium hydroxide alkalinity on SNP and the subsequent formation of the colored nitrosyl-ROS derivative by the attack at position 6 of ROS.

Determination of two antibacterial binary mixtures by chemometrics-assisted spectrophotometry.

Simple chemometrics-assisted spectrophotometric methods are described for determination of 2 antibacterial binary mixtures. The mixtures are composed of norfloxacin in combination with tinidazole and erythromycin (as ethylsuccinate ester or stearate salt) in combination with trimethoprim. The normal UV absorption spectra of each pair of drugs in the studied mixtures, in the range of 200-400 nm, showed a considerable degree of spectral overlapping: 77.5% for the norfloxacin-tinidazole mixture and 84.3% for the erythromycin-trimethoprim mixture. Resolution of the norfloxacin-tinidazole mixture and trimethoprim in the presence of erythromycin was accomplished successfully by using zero-crossing first derivative (1D), classical least-squares (CLS) regression analysis, and principal component regression (PCR) analysis methods. In addition, an alternative simple and accurate colorimetric method was developed for the determination of erythromycin in the presence of trimethoprim using 2,4-dinitrophenylhydrazine. All variables affecting the development of the colored chromogen were studied and optimized, and the product was measured at 526-529 and 538-542 nm for erythromycin stearate and erythromycin ethylsuccinate, respectively. For zero-crossing, first derivative technique Beer's law was obeyed in the general concentration range of 2-50 microg/mL for norfloxacin, tinidazole, and trimethoprim with good correlation coefficients (0.9994-0.9996). Overall limits of detection (LOD) and quantification (LOQ) ranged from 0.59 to 2.81 and 1.96 to 9.33 microg/mL, respectively. The obtained results from CLS and PCR were compared with those obtained from a 1D spectrophotometric method. With the exception of erythromycin, overall recoveries in the average range of 97.33-103.0% were obtained with a considerable degree of accuracy when the suggested methods were applied to analysis of synthetic binary mixtures, some commercial dosage forms such as tablets and oral suspension without interference from the commonly encountered excipients and additives. For the colorimetric method, Beer's law was obeyed in the general concentration range of 7.21-28.84 microg/mL erythromycin with good correlation coefficients (0.9980-0.9996). Overall LOD and LOQ ranged from 0.73 to 1.65 and 2.43-5.49 microg/mL, respectively. Erythromycin derivatives were determined in the commercial dosage form, without interference from trimethoprim-encountered excipients and additives. The obtained results, with both chemometric and colorimetric methods, have been compared with those obtained from reported methods, and proper F- and t-values were observed, indicating no significant difference between the results of the suggested methods and reported method(s). The good percentage recoveries and proper statistical data obtained proved the efficiency of the proposed procedures for the determination of the studied drugs in their binary mixtures as well as in the commercial dosage forms with quite satisfactory precision.

Spectrophotometric analysis of selective serotonin reuptake inhibitors based on formation of charge-transfer complexes with tetracyanoquinodimethane and chloranilic acid.

A simple, accurate, and sensitive spectrophotometric method for analysis of selective serotonin reuptake inhibitors (SSRIs) has been developed and validated. The analysis was based on the formation of colored charge-transfer complexes between the intact molecule of SSRI drug as an n-electron donor and each of tetracyanoquinodimethane (TCNQ) or p-chloranilic acid (pCA) as electron acceptors. The formed complexes were measured spectrophotometrically at 842 and 520 nm for TCNQ and pCA, respectively. Different variables and parameters affecting the reactions were studied and optimized. Under the optimum reaction conditions, linear relationships with good correlation coefficients (0.9975-0.9996) were found between the absorbances and the concentrations of the investigated drugs in the concentration ranges of 4-50 and 20-400 microg/mL with TCNQ and pCA, respectively. With all the investigated drugs, TCNQ gave more sensitive assays than pCA; the limits of assay detection were 2.5-4.8 and 20-40 microg/mL with TCNQ and pCA, respectively. The intra- and interassay precisions were satisfactory; the relative standard deviations did not exceed 2%. The proposed procedures were successfully applied to the analysis of the studied drugs in pure form and pharmaceutical formulations with good accuracy; the recovery values were 98.4-102.8 +/- 1.24-1.81%. The results obtained from the proposed method were statistically comparable with those obtained from the previously reported methods.

Generic nonextractive spectrophotometric method for determination of 4-quinolone antibiotics by formation of ion-pair complexes with beta-naphthol.

This paper describes the development of a generic spontaneous nonextractive spectrophotometric method for determination of 13 pharmaceutically important 4-quinolone antibiotics. The method was based on the formation of yellow-colored water-soluble ion-pair complexes between 2% (w/v) beta-naphthol reagent and each of the studied drugs in sulfuric acid medium at room temperature. The formed ion-pair chromogens have maximum absorption peaks in the range of 365-391 nm. The concentrations of the reagents and the experimental conditions affecting the reaction were optimized. Under the optimum conditions, linear relationships with good linear coefficients (0.9987-0.9995) were found between the absorbance and concentration of the investigated drugs in the range of 10-350 microg/mL. The assay limits of detection and quantitation were 1-9.9 and 3.4-32.9 microg/mL, respectively. The precision of the method was satisfactory; the values of relative standard deviations did not exceed 2%. The proposed method was successfully applied to the analysis of the investigated drugs in pure and pharmaceutical dosage forms with good accuracy and precision; the percentages of label claim ranged from 97.8-102.8 +/- 0.35-1.60%. The results obtained by the proposed spectrophotometric method were comparable with those obtained by the official or reported methods. The proposed method is superior to all the previously reported ion-pair formation-based methods in terms of simplicity because it did not involve extraction procedures for the ion-pair complex. Therefore, this method might be recommended for routine use in quality control laboratories for analysis of the investigated 4-quinolone antibiotics in their pure forms, as well as in pharmaceutical dosage forms.