Green Easily Implemented Spectrophotometric Methods for Concurrent Determination of Ephedrine Hydrochloride and Naphazoline Nitrate in Nasal Preparations Containing Methylparaben.

BACKGROUND: Spectrophotometric resolution of a mixture of several drugs is considered a cheaper, simpler, and more versatile alternative compared to costly chromatographic instruments. OBJECTIVE: The work aims to resolve the interfering spectra of ephedrine hydrochloride, naphazoline nitrate, and methylparaben in nasal preparations using smart spectrophotometric methods. METHOD: In our work, derivative and dual-wavelength methods were combined to eliminate this interference, under the name of derivative dual-wavelength method. Other methods, namely successive derivative subtraction and chemometric analysis, were also able to eliminate this interference. The methods have proven their applicability as they follow the International Conference on Harmonization (ICH) requirements regarding repeatability, precision, accuracy, selectivity, and linearity. Eco-scale, GAPI, and AGREE tools were used to estimate the possible environmental effects of the methods. RESULTS: Acceptable results for repeatability, precision, accuracy, selectivity, and linearity were obtained. Limit of detection (LOD) values were 2.2 for ephedrine and 0.3 for naphazoline. The correlation coefficients were above 0.999. The methods were proven to be safe for application. CONCLUSIONS: The introduced methods are cheap and easily implemented compared to chromatographic techniques. They can be used in purity-checking of raw material and estimation of concentrations in market formulations. The replacement of the published chromatographic techniques with our developed methods is useful when needing to save money, effort, and time. HIGHLIGHTS: The three components of a decongestant nasal preparation were determined using cheap, green, and versatile spectrophotometric methods that keep the advantages of chromatographic techniques, including accuracy, reproducibility, and selectivity.

The Development of Spectrophotometric and Validated Stability- Indicating RP-HPLC Methods for Simultaneous Determination of Ephedrine HCL, Naphazoline HCL, Antazoline HCL, and Chlorobutanol in Pharmaceutical Pomade Form.

BACKGROUND: Allergic rhinitis, acute nasal congestion and sinusitis are one of the most common health problems and have a major effect on the quality of life. Several medications are used to improve the symptoms of such diseases in humans. Pharmaceutical pomade form containing Ephedrine (EPD) HCl, Naphazoline (NPZ) HCl, Antazoline (ANT) HCl, and Chlorobutanol (CLO) is one of them. OBJECTIVE: For these reasons, this study includes the development of spectrophotometric and chromatographic methods for the determination of EPD HCl, NPZ HCl, ANT HCl, and CLO active agents in the pharmaceutical pomade. METHOD: In the spectrophotometric method, third-order derivative of the amplitudes at 218 nm n=5 and the first-order derivative of the amplitudes 254 nm n=13 was selected for the determination of EPD, ANT, respectively while NPZ was determined by the second derivative at 234 nm and n=21. Colorimetric detection was applied for assay analysis of CLO at 540 nm. Furthermore, a reverse phase high performance liquid chromatographic (RP- HPLC) method has been developed and optimized by using Agilent Zorbax Eclipse XDB C18 (75 mm x 3.0 mm, 3.5µm) column. The column temperature was 40°C, binary gradient elution was used and the mobile phase consisted of 15 mM phosphate buffer in distilled water (pH 3.0) and methanol, and the flow rate was 0.6 mL min-1 and the UV detector was detected at 210 nm. The linear operating range was obtained as 11.97-70, 0.59-3, 2.79-30, and 2.92-200 µg mL-1 for EPD HCl, NPZ HCl, ANT HCl, and CLO respectively. RESULTS: The LOD values were found to be 3.95, 0.19, 0.92 and 0.96 µg mL-1 for EPD HCl, NPZ HCl, ANT HCl, and CLO in the spectrophotometric method, respectively. The linear ranges in the RP-HPLC method were 8.2-24.36 µg mL-1, 0.083 - 0.75 µg/mL, 2.01-7.5 µg mL-1 and 2.89-24.4 µg mL-1 for EPD HCl, NPZ HCl, ANT HCl, and CLO, respectively. The LOD values in the validation studies were 2.7, 0.025, 0.66 and 0.86 µg mL-1 for EPD HCl, NPZ HCl, ANT HCl, and CLO in RP-HPLC method respectively. CONCLUSION: The results of the spectrophotometric and chromatographic methods were compared and no differences were found between the two methods.

A sub-minute electrophoretic method for simultaneous determination of naphazoline and zinc.

This paper reports for the first time, a method for simultaneous determination of naphazoline (NPZ) and zinc (Zn) using an analytical separation technique (capillary electrophoresis with capacitively coupled contactless conductivity detection -CE-C4D). A single run is possible every 55s (sampling rate=65h-1). The separation by CE-C4D was achieved on a fused silica capillary (50cm length - 10cm effective, 50µm i.d.) with a background electrolyte (BGE) composed by 20mmolL-1 of 2-(morpholin-4-yl)ethane-1-sulfonic acid (MES) and 20mmolL-1 of histidine (HIS) (pH 6.0). Detection limits were estimated at 20 and 30µmolL-1 and recovery values for spiked samples were 98 and 102% for NPZ and Zn, respectively. The developed procedure was compared to HPLC (NPZ) and FAAS (Zn) and no statistically significant differences were observed (95% confidence level).

Simultaneous determination of chloramphenicol, dexamethasone and naphazoline in ternary and quaternary mixtures by RP-HPLC, derivative and wavelet transforms of UV ratio spectra.

The application of chemometrics-assisted UV spectrophotometry and RP-HPLC to the simultaneous determination of chloramphenicol, dexamethasone and naphazoline in ternary and quaternary mixtures is presented. The spectrophotometric procedure is based on the first-order derivative and wavelet transforms of ratio spectra using single, double and successive divisors. The ratio spectra were differentiated and smoothed using Savitzky-Golay filter; whereas wavelet transform realized with wavelet functions (i.e. db6, gaus5 and coif3) to obtain highest spectral recoveries. For the RP-HPLC procedure, the separation was achieved on a ZORBAX SB-C18 (150×4.6 mm; 5 µm) column at ambient temperature and the total run time was less than 7 min. A mixture of acetonitrile - 25 mM phosphate buffer pH 3 (27:73, v/v) was used as the mobile phase at a flow rate of 1.0 mL/min and the effluent monitored by measuring absorbance at 220 nm. Calibration graphs were established in the range 20-70 mg/L for chloramphenicol, 6-14 mg/L for dexamethasone and 3-8 mg/L for naphazoline (R(2)>0.990). The RP-HPLC and ratio spectra transformed by a combination of derivative-wavelet algorithms proved to be able to successfully determine all analytes in commercial eye drop formulations without sample matrix interference (mean percent recoveries, 97.4-104.3%).

Determination of naphazoline hydrochloride in biological and pharmaceutical samples by a quantum dot-assisted chemiluminescence system using response-surface methodology.

A simple and highly sensitive chemiluminescence (CL) method is reported for the determination of naphazoline hydrochloride (NH). It was found that the weak CL from the reaction of luminol and KIO4 in an alkaline medium could be highly amplified by cysteine-capped cadmium telluride quantum dots (QDs) and the enhanced CL was effectively quenched by NH and this finding was utilized as a basis for the determination of NH. The QDs were synthesized in aqueous medium and characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), and UV-vis and photoluminescence spectroscopy. A possible mechanism was proposed for the CL system based on radical identification experiments, along with CL spectrum of the system. The experimental parameters were optimized by the reliable response surface methodology (RSM). Under the optimized experimental conditions, the proposed method allowed the determination of NH over the range of 5.0 × 10(-10) -2.0 × 10(-7) mol/L (r(2) = 0.9993, n = 10). The precision (RSD%) of the method, obtained from five replicate determinations of 2.0 and 150 nmol/L NH, was found to be 1.0% and 1.3%, respectively. The method was successfully applied to the determination of NH in pharmaceutical formulations and human urine and serum samples with results corroborated with the aid of those obtained from a standard method.

Flow injection chemiluminescence determination of naphazoline hydrochloride in pharmaceuticals.

A simple and sensitive flow injection chemiluminescence (FI-CL) method was developed for the determination of naphazoline hydrochloride (NPZ). The method is based on the enhancing effect of NPZ on the weak CL signal from the reaction of KIO4 with H2 O2 . Experimental parameters that affected the CL signal, including the pH of the KIO4 solution, concentrations of KIO4 , H2 O2 and disodium-EDTA and flow rate were optimized. Under the optimum conditions, the increment of CL intensity was linearly proportional to the concentration of NPZ in the range 5.0 × 10(-6) to 70 × 10(-6) mol/L. The detection limit was 1.0 × 10(-6) mol/L and the relative standard deviation for 50 × 10(-6) mol/L NPZ solution was 2.8% (n = 11). In addition, a high throughput of 120 samples/h was achieved. The utility of this method was demonstrated by determining NPZ in pharmaceuticals.

Report: Simultaneous determination of naphazoline hydrochloride, chlorpheniramine maleate and methylene blue in their ternary mixture.

Validated spectrophotometric and chemometric methods were developed for determination of Naphazoline Hydrochloride (NAP), Chlorpheniramine maleate (CLO) and Methylene blue (MB) in their ternary mixture. Method A was a spectrophotometric method, where NAP and MB were determined using second derivative (D²) spectrophoto metric method using the peak amplitudes at 299 nm and 337 nm for NAP and MB respectively , while CLO was determined using second derivative ratio (DD²) spectrophotometric method using the peak amplitude at 276.6 nm. Method B used the chemometric techniques; principal component regression (PCR) and partial least squares (PLS) for determination of NAP, CLO and MB using the information contained in the absorption spectra of their ternary mixture solutions. The proposed methods have been successfully applied for the analysis of NAP, CLO and MB in their pharmaceutical formulation and the obtained results were statistically compared with the reported methods.

Ionic liquids as mobile phase additives for feasible assay of naphazoline in pharmaceutical formulation by HPTLC-UV-densitometric method.

A specific and reliable high-performance thin layer chromatography method with densitometry detection has been developed for the determination of naphazoline nitrate in nasal drops. The best separation of the basic analyte, without spot tailing, was achieved by using a mobile phase composed of acetonitrile-water (60:40, v/v), adding 1.5 % (v/v) imidazolium-class ionic liquid and covering the plates with a stationary phase based on RP-18 with F254S (10 × 20 cm). The presented results confirm that imidazolium tetrafluoroborate ionic liquids are efficient suppressors of free silanols, which are considered to be responsible for troublesome and irreproducible chromatographic determinations of basic compounds. The developed chromatographic system was found to be convenient in use and to provide a repeatable assay of naphazoline nitrate in nasal drops, which could not be obtained with the use of standard silanol suppressing mobile phase additives such as triethylamine or dimethyloctylamine.

Construction and performance characterization of screen printed and carbon paste ion selective electrodes for potentiometric determination of naphazoline hydrochloride in pharmaceutical preparations.

This paper describes the development of screen-printed (SPE) and carbon paste (CPE) sensors for the rapid and sensitive quantification of naphazoline hydrochloride (NPZ) in pharmaceutical formulations. This work compares the electroactivity of conventional carbon paste and screen-printed carbon paste electrodes towards potentiometric titration of NPZ. The repeatability and accuracy of measurements performed in the analysis of these pharmaceutical matrices using new screen printed sensors were evaluated. The influence of the electrode composition, conditioning time of the electrode and pH of the test solution, on the electrode performance were investigated. The drug electrode showed Nernstain responses in the concentration range from 1 × 10(-6) to 1 × 10(-2) mol L(-1) with slopes of 57.5 ± 1.3 and 55.9 ± 1.6 mV per decade for SPE and CPE, respectively, and was found to be very precise and usable within the pH range 3-8. These sensors exhibited a fast response time (about 3 s for both SPE and CPE, respectively), a low detection limit (3.5 × 10(-6) and 1.5 × 10(-6) M for SPE and CPE, respectively), a long lifetime (3 and 2 months for SPE and CPE, respectively) and good stability. The selectivity of the electrode toward a large number of inorganic cations, sugars and amino acids was tested. It was applied to potentiometric determination of NPZ in pure state and pharmaceutical preparation under batch conditions. The percentage recovery values for the assay of NPZ in tablets (relative standard deviations ≤0.3% for n = 4) were compared well with those obtained by the official method.

Simultaneous determination of antazoline and naphazoline by the net analyte signal standard addition method and spectrophotometric technique.

A novel net analyte signal standard addition method (NASSAM) was used for simultaneous determination of the drugs anthazoline and naphazoline. The NASSAM can be applied for determination of analytes in the presence of known interferents. The proposed method is used to eliminate the calibration and prediction steps of multivariate calibration methods; the determination is carried out in a single step for each analyte. The accuracy of the predictions against the H-point standard addition method is independent of the shape of the analyte and interferent spectra. The net analyte signal concept was also used to calculate multivariate analytical figures of merit, such as LOD, selectivity, and sensitivity. The method was successfully applied to the simultaneous determination of anthazoline and naphazoline in a commercial eye drop sample.

Flow-injection chemiluminescence method for the determination of naphazoline hydrochloride and oxymetazoline hydrochloride.

A sensitive and simple flow-injection chemiluminescence (FI-CL) method, which was based on the CL intensity generated from the redoxreaction of potassium permanganate (KMnO4)-formaldehyde in vitriol (H2SO4) medium, has been developed, validated and applied for the determination of naphazoline hydrochloride and oxymetazoline hydrochloride. Besides oxidants and sensitizers, the effect of the concentration of H(2)SO(4), KMnO4 and formaldehyde was investigated. Under the optimum conditions, the linear range was 1.0 x 10(-2)-7.0 mg/L for naphazoline hydrochloride and 5.0 x 10(-2)-10.0 mg/L for oxymetazoline hydrochloride. During seven repeated inter-day and intra-day precision tests of 0.1, 1.0 and 10.0 mg/L samples, the relative standard deviations all corresponded to reference values. The detection limit was 8.69 x 10(-3) mg/L for naphazoline hydrochloride and 3.47 x 10(-2) mg/L for oxymetazoline hydrochloride (signal-to-noise ratio < or = 3). This method has been successfully implemented for the determination of naphazoline hydrochloride and oxymetazoline hydrochloride in pharmaceuticals.

A rapid derivative spectrophotometric method for simultaneous determination of naphazoline and antazoline in eye drops.

A zero-crossing first-derivative spectrophotometric method is applied for the simultaneous determination of naphazoline hydrochloride and antazoline phosphate in eye drops. The measurements were carried out at wavelengths of 225 and 252 nm for naphazoline hydrochloride and antazoline phosphate, respectively. The method was found to be linear (r2>0.999) in the range of 0.2-1 microg/ml for naphazoline hydrochloride in the presence of 5 microg/ml antazoline phosphate at 225 nm. The same linear correlation (r2>0.999) was obtained in the range of 1-10 microg/ml of antazoline phosphate in the presence of 0.5 microg/ml of naphazoline hydrochloride at 252 nm. The limit of determination was 0.2 microg/ml and 1 microg/ml for naphazoline hydrochloride and antazoline phosphate, respectively. The method was successfully used for simultaneous analysis of naphazoline hydrochloride and antazoline phosphate in eye drops without any interference from excipients and prior separation before analysis.

A fluorescence optosensor for analyzing naphazoline in pharmaceutical preparations. Comparison with other sensors.

We have developed an optical sensor for determining and quantifying naphazoline (NPZ) based on its inherent fluorescence property. We have placed a non-ionic-exchanger solid support (Amberlite XAD-7) in a flow cell in the light path of the excitation beam and the fluorescence signal for NPZ is continuously monitored at lambda(exc/nm)=294/306 nm. The response time for this sensor is acceptably fast, 80s, obtaining a detection limit of 2.6 ng mL(-1) with standard deviations of 2.0% at 125 ng mL(-1). This device has been satisfactorily applied to two commercial formulations and its selectivity has been demonstrated with an interference study. The advantages have been compared with the only published sensor for determining NPZ in pharmaceutical preparations and with other analytical methods in the literature.

Comparison of three different phosphorescent methodologies in solution for the analysis of naphazoline in pharmaceutical preparations.

We present results from a comparative study of three proposed phosphorimetric methods for determination of naphazoline (NPZ) in solution. The first method is based on use of micelles to stabilize phosphorescence signals in solutions at room temperature (MS-RTP). The second is based on the use of a heavy atom salt and sodium sulfite as an oxygen scavenger to obtain room-temperature phosphorescence (HAI-RTP) in solution. The last method employs an optical sensor for NPZ based on the phosphorescent properties of the analyte on a solid sensor phase. The aim of this work was to compare time consumption, simplicity, sensitivity, selectivity, detection, and quantification limits for use of these three phosphorimetric methods to determine naphazoline in pharmaceutical preparations. The most simple, sensitive, and reproducible of the three methods for naphazoline analysis is the HAI-RTP method. Detection limits are 4.9, 1.7, and 9.4 ng mL(-1), respectively, for the MS-RTP, HAI-RTP, and optosensor methods.

Simultaneous determination of naphazoline, diphenhydramine and phenylephrine in nasal solutions by capillary electrophoresis.

A capillary zone electrophoresis (CZE) method has been developed to separate and quantitate naphazoline (NAPH), dyphenhydramine (DIP) and phenylephrine (PHE) in nasal solutions. Samples were diluted 1:25 in ultrapure water and injected at the anodic end. A central composite design has been used to optimise the experimental conditions for a complete and fast separation of the active ingredients studied. Critical parameters such as voltage, pH and buffer concentration have been studied to evaluate how they affect responses such as resolution and migration times. Separation was performed on a silica capillary with 75 microm I.D. and 70 cm total length at an applied voltage of 17.7 kV with a phosphate run buffer of pH 3.72 and 0.063 mol l(-1). Calibration curves were prepared for NAPH, DIP and PHE. For each analyte, the correlation coefficients were >0.999 (n=15). The RSD% of six replicate injections for each analyte were reasonably good. The method was applied to the quantitation of the three components in a commercial dosage form. The proposed method has the advantage of needing a very simple sample pretreatment and being faster than a typical HPLC chromatographic method.

Complementary use of partial least-squares and artificial neural networks for the non-linear spectrophotometric analysis of pharmaceutical samples.

The complementary use of partial least-squares (PLS) multivariate calibration and artificial neural networks (ANNs) for the simultaneous spectrophotometric determination of three active components in a pharmaceutical formulation has been explored. The presence of non-linearities caused by chemical interactions was confirmed by a recently discussed methodology based on Mallows augmented partial residual plots. Ternary mixtures of chlorpheniramine, naphazoline and dexamethasone in a matrix of excipients have been resolved by using PLS for the two major analytes (chlorpheniramine and naphazoline) and ANNs for the minor one (dexamethasone). Notwithstanding the large number of constituents, their high degree of spectral overlap and the occurrence of non-linearities, rapid and simultaneous analysis has been achieved, with reasonably good accuracy and precision. No extraction procedures using non-aqueous solvents are required.

[Applications of phosphorimetry in pharmaceutical analysis].

Applications of phosphorimetry including solid substrate phosphorescence, liquid medium phosphorescence, low temperature phosphorescence and phosphorescence sensors were reviewed in pharmaceutical analysis. The drugs involved here included the varieties of alkaloid, Chinese traditional medicine, tetracyclines, quinolone, riboflavin, anticancer medicine, naphazoline, naproxen, nafronyl dipyridamole and so on. Solid surface phosphorimetry is characterized by sample volume of microliter grade, simple and fast operation procedures in pharmaceutical analysis. The combination of liquid phosphorescence with flow injection analysis and chemosensing technique has good advantages in fast, continuous and on-line monitoring of medicines. Modified low temperature phosphorimetry still remains its high sensitivity and overcomes some disadvantages in the procedures. Phosphorimetry will be more widely applied to pharmaceutical analysis as the development of sensitive and quenching, energy transfer, derivative and immunization luminescence.

Two examples of rapid and simple drug analysis in pharmaceutical formulations using capillary electrophoresis: naphazoline, dexamethasone and benzalkonium in nose drops and nystatin in an oily suspension.

Capillary electrophoresis is a versatile tool in pharmaceutical analysis. In the course of a revision of the "Standardrezepturen", a German formula of standard dispensings for preparation in pharmacies, this technique has been applied to drug analysis in pharmaceutical formulations. The present paper deals with two different examples. First, naphazoline, dexamethasone and the preservative benzalkonium are quantified in nose drops without any sample preparation. Second, the antifungal antibiotic nystatin is quantified using nonaqueous capillary electrophoresis in methanol after sample preparation from an oily suspension.

Room-temperature phosphorimetric method for the determination of the drug naphazoline in pharmaceutical preparations.

A selective and sensitive micelle-stabilized room-temperature phosphorimetric (MS-RTP) method for the determination of naphazoline in pharmaceutical preparations is described. The method is based on obtaining a phosphorescence signal from naphazoline using a micellar agent (sodium dodecyl sulfate), a heavy atom salt (TINO3) and a deoxygenation agent (Na2SO3). Optimization of the various conditions permitted the establishment of an MS-RTP method for naphazoline determination with a detection limit of 64.2 ng ml-1 and a relative standard deviation of 3.74% at the 500 ng ml-1 level. The method was applied to the analysis of pharmaceutical preparations.