Sequential injection analysis for the determination of fluoroquinolone antibacterial drug residues by using eosin Y as complexing agent.

A sequential injection analysis (SIA) method was developed for the rapid and sensitive determination of fluoroquinolone residues, including norfloxacin, ciprofloxacin and enrofloxacin, in fish samples. The method is based on the reaction between fluoroquinolone drug and eosin Y in Britton-Robinson buffer (pH 2.0), forming pink colored complexs with maximum absorptions at 522, 525 and 527 nm for norfloxacin, ciprofloxacin and enrofloxacin, respectively. Linearity ranges were found to be 0.05-10.0 mg L-1 (r2 = 0.9996), 0.1-10.0 mg L-1 (r2 = 0.9995) and 0.05-10.0 mg L-1 (r2 = 0.9997) for norfloxacin, ciprofloxacin and enrofloxacin, respectively. The detection limit was found to be in the range of 0.013-0.019 mg L-1. The method was tested and validated for various parameters according to main guidelines. The proposed SIA method was successfully applied for the determination of fluoroquinolone drug residues in fish samples with the sampling rate of 47 h-1. The results demonstrated that the method is accurate, precise and reproducible, while being simple, rapid, economical and less time consuming. It can be suitably applied for the estimation of fluoroquinolone drug residues in routine quality control.

The Isolation of Lutein and Lutein 3'-methyl ether from Peristrophe lanceolaria.

Two carotenoids, lutein (1) and lutein 3'-methyl ether (2), have been isolated from the EtOAc fraction othe MeOH extract of Peristrophe lanceolaria, growing in Thailand. The structures of these compounds were elucidated from their ID and 2D NMR spectroscopic data and from comparisons made with the literature data. This is the first report of the isolation of lutein-3'-methyl ether as a natural product.

A new 1,6-benzoxazocine-5-one alkaloid isolated from the aerial parts of Peristrophe lanceolaria.

A new 1,6-benzoxazocine-5-one alkaloid has been isolated as its butyl acetal derivative (1) along with peristrophine from the n-BuOH and EtOAc fractions of the crude MeOH extract of the aerial parts of Peristrophe lanceolaria growing in Thailand. The structures of these compounds were elucidated on the basis of their spectroscopic data. These compounds were isolated for the first time from P. lanceolaria. The EtOAc and n-BuOH fractions also possessed significant antioxidant activity with IC50 values of 57 and 50 µg/mL, respectively (DPPH method), whereas 1 had an IC50 value of 23 µg/mL.

Determination of gallic acid with rhodanine by reverse flow injection analysis using simplex optimization.

A reversed flow injection (rFI) system was designed and constructed for gallic acid determination. Gallic acid was determined based on the formation of chromogen between gallic acid and rhodanine, resulting in a colored product with a λmax at 520 nm. The optimum conditions for determining gallic acid were also investigated. Optimizations of the experimental conditions were carried out based on the so-call univariate method. The conditions obtained were 0.6% (w/v) rhodanine, 70% (v/v) ethanol, 0.9 mol L(-1) NaOH, 2.0 mL min(-1) flow rate, 75 µL injection loop and 600 cm mixing tubing length, respectively. Comparative optimizations of the experimental conditions were also carried out by multivariate or simplex optimization method. The conditions obtained were 1.2% (w/v) rhodanine, 70% (v/v) ethanol, 1.2 mol L(-1) NaOH, flow rate 2.5 mL min(-1), 75 µL injection loop and 600 cm mixing tubing length, respectively. It was found that the optimum conditions obtained by the former optimization method were mostly similar to those obtained by the latter method. The linear relationship between peak height and the concentration of gallic acid was obtained over the range of 0.1-35.0 mg L(-1) with the detection limit 0.081 mg L(-1). The relative standard deviations were found to be in the ranges 0.46-1.96% for 1, 10, 30 mg L(-1) of gallic acid (n=11). The method has the advantages of simplicity extremely high selectivity and high precision. The proposed method was successfully applied to the determination of gallic acid in longan samples without interferent effects from other common phenolic compounds that might be present in the longan samples collected in northern Thailand.

Concise synthesis of (-)-steviamine and analogues and their glycosidase inhibitory activities.

A concise synthesis of (-)-steviamine is reported along with the synthesis of its analogues 10-nor-steviamine, 10-nor-ent-steviamine and 5-epi-ent-steviamine. These compounds were tested against twelve glycosidases (at 143 µg mL(-1) concentrations) and were found to have in general poor inhibitory activity against most enzymes. The 10-nor analogues however, showed 50-54% inhibition of α-L-rhamnosidase from Penicillium decumbens while one of these, 10-nor-steviamine, showed 51% inhibition of N-acetyl-ß-D-glucosaminidase (from Jack bean) at the same concentration (760 µM).

Chemical constituents and antioxidant and biological activities of the essential oil from leaves of Solanum spirale.

The essential oil of the leaves Solanium spirale Roxb. was isolated by hydrodistillation and analyzed for the first time using GC and GC-MS. Thirty-nine constituents were identified, constituting 73.36% of the total chromatographical oil components. (E)-Phytol (48.10%), n-hexadecanoic acid (7.34%), beta-selinene (3.67%), alpha-selinene (2.74%), octadecanoic acid (2.12%) and hexahydrofarnesyl acetone (2.00%) were the major components of this oil. The antioxidant activity of the essential oil was evaluated by using the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging assay. The oil exhibited week antioxidant activity with an IC50 of 41.89 mg/mL. The essential oil showed significant antibacterial activity against both Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus with MIC values of 43.0 microg/mL and 21.5 microg/mL, respectively. It also showed significant cytotoxicity against KB (oral cancer), MCF-7 (breast cancer) and NCI-H187 (small cell lung cancer) with the IC50 values of 26.42, 19.69, and 24.02 microg/mL, respectively.

The isolation of bioactive flavonoids from Jacaranda obtusifolia H. B. K. ssp. rhombifolia (G. F. W. Meijer) Gentry.

The paper describes the bioassay-guided isolation, structure elucidation and anticancer evaluation of five flavonoids (-)-liquiritigenin (1), (-)-neoliquiritin (2), isoliquiritigenin (3), isoliquiritin (4) and formononetin (5) from the twigs of Jacaranda obtusifolia H. B. K. ssp. rhombifolia (G. F. W. Meijer) Gentry. The structures were elucidated based on ¹H, ¹³C NMR, comprehensive 2D NMR, MS analyses and comparison with previously reported spectral data. Compounds 1 and 3 were demonstrated to be inhibitory in vitro against NCI-H187 (small cell lung cancer) with IC50 values of 30.1 and 16.6 µg mL⁻¹, respectively. The isolates were non-cytotoxic to Vero cells (African green monkey kidney).

Sequential injection spectrophotometric determination of tetracycline antibiotics in pharmaceutical preparations and their residues in honey and milk samples using yttrium (III) and cationic surfactant.

A sequential injection analysis (SIA) spectrophotometric method for determining tetracycline (TC), chlortetracycline (CTC) and oxytetracycline (OTC) in different sample matrices were described. The method was based on the reaction between tetracyclines and yttrium (III) in weak basic micellar medium, yielding the light yellow complexes, which were monitored at 390, 392 and 395 nm, respectively. A cationic surfactant, cetyltrimethylammonium bromide (CTAB) was used to obtain the micellar system. The linear ranges of calibration graphs were between 1.0 × 10(-5) and 4 × 10(-4) mol L(-1), respectively. The molar absorptivities were 5.24 × 10(5), 4.98 × 10(4) and 4.78 × 10(4) L mol(-1)cm(-1). The detection limits (3σ) were between 4.9 × 10(-6) and 7.8 × 10(-6) mol L(-1) whereas the limit of quantitations (10σ) were between 1.63 × 10(-5) and 2.60 × 10(-5) mol L(-1) the interday and intraday precisions within a weak revealed as the relative standard deviations (R.S.D., n=11) were less than 4%. The method was rapid with a sampling rate of over 60 samples h(-1) for the three drugs. The proposed method has been satisfactorily applied for the determination of tetracycline and its derivatives in pharmaceutical preparations together with their residues in milk and honey samples collected in Chiang Mai Province. The accuracy was found to be high as the Student's t-values were found to be less than the theoretical ones. The results were compared favorably with those obtained by the conventional spectrophotometric method.

High-performance thin-layer chromatographic determination of ketoconazole in pharmaceutical formulations.

A high-performance thin-layer chromatographic method was developed for the determination of ketoconazole. The sample was separated on a silica gel 60 F254 plate and developed in ethanol-acetone-1.0 mol l-1 H2SO4 by means of an automatic multiple-development system. The area of the spot was quantified by a TLC scanner at a wavelength of 298 nm. A linear calibration curve was established over the range of 3-20 µg/ml of ketoconazole, with a correlation coefficient of 0.9992. The relative standard deviations for intraday and interday precisions, for three replicate determinations, were found to be 1.72% and 0.69% for 5 µg/ml and 2.18% and 0.94% for 10 µg/ml of ketoconazole, respectively. The average percentage recoveries of ketoconazole shampoos (Nora, Kenalyn, and Nizoral) and ketoconazole creams (Nizoral, Fungasin, and Ketazon) were found to be 96.10, 97.06, and 99.58, and 96.77, 97.26, and 95.74, respectively. This method has been applied to the determination of ketoconazole in various pharmaceutical dosage forms. Common excipients in formulations do not interfere. This method is simple, precise, accurate, and inexpensive. It should be used for routine analysis.

Sequential injection analysis with lab-at-valve (SI-LAV) for the determination of solasodine in Solanum species.

The development of sequential injection analysis with lab-at-valve (LAV) semi-automated system on-line liquid-liquid extraction is demonstrated for spectrophotometric determination of solasodine in various Solanum species fruits. The main proposed is semi-automated extractive determination of solasodine using methyl orange as colorimetric reagent. After optimization of the system, sample, reagent and organic solvent were sequentially aspirated into an extraction coil connected to the center of a selection valve, where extraction took place by flow reversal. The aqueous and organic phases were separated in a lab-at-valve unit attracted to one of the ports of the selection valve. The absorption of ion-pair solasodine-methyl orange complex in the organic phase was measured spectrophotometrically at 420 nm. The method performances, including reproducibility, linearity, sensitivity and accuracy, were also evaluated. The proposed method is simple, reproducible and accurate. It was successfully applied to the determination of solasodine in Solanum aculeatissimum Jacq., Solanum violaceum Ortega., Solanum melongena Linn. and Solanum indicum Linn. fruits in Solanaceae family. Results obtained were in good agreement with those obtained by batch wise spectrophotometric method. It is also suitable and useful for determination of solasodine in other medicinal plants.

A cleaner and simple spectrophotometric micro-fluidic procedure for copper determination using nitroso-R salt as chromogenic agent.

A cleaner and simple spectrophotometric method using microflow analysis (muFA) was performed. It consisted of a T-junction with microcoil on a polymethyl methacrylate (PMMA) chip which was fabricated by laser ablation and a molded polydimethylsiloxane (PDMS) as top plate. The fabricated PMMA chip was integrated with light emitting diode (LED) as light source and spectrometer as detector. The proposed device was applied to determining copper in water samples using nitroso-R salt as chromogenic reagent at 495 nm. It was found that the proposed muFA system was with less reagents and samples consumption with tiny waste generation. The relative standard deviation (R.S.D.) was less than 2% (n=11) with the percentage recovery of 98.0+/-1.7% (n=7). The linear range for determination of copper in water samples was over the range of 0.05-3.0 microg mL(-1) with a correlation coefficient (r2) of 0.999. The limit of detection (3sigma) was 47 ng mL(-1) with a sample throughput of 30 h(-1).

High-performance liquid chromatographic determination of arbutin in skin-whitening creams and medicinal plant extracts.

A high-performance liquid chromatographic method was developed for quantitative analysis of arbutin. The arbutin was separated on an ODS Hypersil C(18) column with a mobile phase of water:methanol:0.1 M hydrochloric acid (89:10:1, v/v/v). The level of arbutin was measured by means of UV detection at 222 nm. The optimum conditions for arbutin quantitative analysis were investigated. The calibration curve was found to be linear up to 1,000 microg/ml(-1) of arbutin concentration, and the working calibration curve for arbutin determination over the range 0.5-30.0 microg/ml(-1) of arbutin (r(2)=0.9999) was established. The relative standard deviations for intraday and interday were found to be 0.98% and 1.15%, respectively. A detection limit (3sigma) and quantitation limit (10sigma) of 0.02 microg/ml(-1) and 0.2 microg/ml(-1), respectively, and a mean percentage recovery of the spiked arbutin of 99.88 +/- 1.12% were obtained. The proposed method has been applied to the determination of arbutin in commercial skin-whitening creams (Arbuwhite cream, Super Whitening cream, and Shiseido cream) with average contents of 7.60, 5.30, and 57.90 mg/g(-1), respectively. It was also applied to the determination of arbutin in medicinal plant extracts from Betula alnoides Buch. Ham., Clerodendrum petasites S. Moore, Curculigo latifolia Dryand. Var. latifolia, and Hesperethusa crenulata (Roxb.) Roem, levels of which were found to be 3.50, 1.50, 1.10, and 0.12 microg/g(-1), respectively (no article reported in the literature about arbutin analysis). The proposed HPLC method is rapid, simple, and selective for routine analysis.

Simultaneous determination of neomycin sulfate and polymyxin B sulfate by capillary electrophoresis with indirect UV detection.

A simple and rapid capillary electrophoresis method, with indirect UV detection, for the simultaneous determination of neomycin sulfate and polymyxin B sulfate in pharmaceutical formulations was developed. Critical parameters such as pH, buffer composition and concentration, voltage and injection time have been studied to evaluate, how they affect responses, such as resolution and migration times. Separation was performed on a fused silica capillary with 50 microm i.d. and 27 cm total length at an applied voltage of 6 kV with a 15 mM phosphate run buffer (pH 5.0) containing 40 mM N-(4-hydroxy-phenyl)acetamide and 50 mM tetradecylammonium bromide (TTAB). The detection wavelength was set at 280 nm. Quantitative analysis was validated by testing the reproducibility of the method, giving a relative standard deviation less than 0.4 and 2.4% for the repeatability of migration time and corrected peak area, respectively. Accuracy was tested by spiking eye-ear formulations with standards and the recoveries of neomycin sulfate and polymyxin B sulfate were found to be between 97.44-103.18% and 96.85-101.68%, respectively. Linearity of neomycin sulfate and polymyxin B sulfate were obtained in the ranges of 17-682 and 24-608 microg/mL, respectively, with r(2) values above 0.999. The established TLC-densitometric method was applied to evaluate the proposed CE method, and comparable results were obtained by using CE with much shorter analysis time and a small quantity of solvents consumed. The developed method is also the first report on the simultaneous determination of neomycin sulfate and polymyxin B sulfate in pharmaceutical preparations by CE.

Sequential injection spectrophotometric determination of zinc(II) in pharmaceuticals based on zinc(II)-PAN in non-ionic surfactant medium.

A sequential injection analysis (SIA) spectrophotometric method for the determination of trace amounts of zinc(II) with 1-(2-pyridylazo)-2-naphthol (PAN) is described. The method is based on the measurement of absorbance of the zinc(II)-PAN chelate solubilized with a non-ionic surfactant, Triton X-100, no extraction procedure is required in the proposed method, yielding a pink colored complex at pH 9.5 with absorption maximum at 553nm. The SIA parameters that affect the signal response have been optimized in order to get the better sensitivity and minimum reagent consumption. A linear relationship between the relative peak height and concentration was obtained in the concentration range of 0.1-1.0microg ml(-1). The limit of detection (LOD, defined as 3sigma) and limit of quantification (LOQ, defined as 10sigma) were 0.02 and 0.06microg ml(-1), respectively. The sample throughput about 40 samples/h was obtained. The repeatability were 1.32 and 1.24% (n=10) for 0.1 and 0.5microg ml(-1), respectively. The proposed method was successfully applied to the assay of zinc(II) in three samples of multivitamin tablets. The results were found to be in good agreement with those obtained by flame atomic absorption spectrophotometric method and with the claimed values by the manufactures. The t-test showed no significant difference at 95% confidence level.

Greener analytical method for the determination of copper(II) in wastewater by micro flow system with optical sensor.

Greener analytical method using micro flow system for the determination of Cu(II) in wastewater samples was designed and investigated. The micro flow system consisted of a planar glass chip with poly(dimethylsiloxane) (PDMS) top plate and fixed with fiber optic probe as optical sensor for monitoring of Cu(II) that reacted with 2-carboxy-2'-hydroxy-5'-sulfoformazyl benzene (zincon) on the chip at 605nm. This design gave a satisfied sensitivity with a linear calibration graph over the range of 0.1-3.0mugmL(-1) of Cu(II) and correlation coefficient 0.9991. The percentage relative standard deviation was 2.5 for 10-replicate measurements and the limit of detection (LOD) was 0.1mugmL(-1). This system has been successfully applied to the determination of Cu(II) in wastewaters from electroplating industry with less reagents and samples consumption and diminutive waste generation.

Determination of cefadroxil by sequential injection with spectrophotometric detector.

A sequential injection analysis (SIA) spectrophotometric procedure for cefadroxil determination has been developed. The SIA instrumentation was modified to achieve the desired function and operations by using the software developed to interface the PC with the conventional SIA system. The method is based on the measurement of a red, water-soluble product formed by the reaction between cefadroxil and 4-aminoantipyrine in the presence of alkaline potassium hexacyanoferrate(III) at 510 nm. Optimum conditions for determining the drug were investigated. Beer's law was obeyed over the concentration ranges of 1 - 10 mg L(-1) and 10 - 50 mg L(-1) with a detection limit (3 sigma) of 0.17 mg L(-1) and a limit of quantification (10 sigma) of 0.56 mg L(-1). The relative standard deviations of 1.98% and 1.93% for 5 mg L(-1) and 30 mg L(-1) of the drug, respectively (n = 11) are obtained. The proposed method has been applied satisfactorily to the determination of cefadroxil in commercial pharmaceutical formulations with a sampling rate of 100 h(-1). Results obtained were in good agreement with those obtained by the official HPLC method at the 95% confidence level.

Flow injection chemiluminescence determination of cefadroxil using potassium permanganate and formaldehyde system.

A simple, rapid and precise flow injection chemiluminescence (FI-CL) method is proposed for the determination of cefadroxil and is suitable for application to other antibiotics containing phenolic hydroxyl groups. A possible mechanism for this selectivity is suggested. The method is based on the CL-emitting reaction between cefadroxil and potassium permanganate in sulfuric acid medium, enhanced by formaldehyde (HCHO). Under the optimum conditions, calibration graphs over the ranges of 0.05-0.8 and 1.0-10.0 microg ml(-1) were obtained. The proposed method was successfully applied to the determination of cefadroxil in pharmaceutical formulations with no evidence of interference from common excipients. The detection limit (3sigma) of this method is 25 ng ml(-1) (6.9 x 10(-8) mol l(-1)). The relative standard deviation was less than 2% for 0.4 and 4.0 microg ml(-1) cefadroxil (n = 20). The sample throughput was found to be 120 h(-1).

Flow injection spectrophotometric determination of tetracycline in a pharmaceutical preparation by complexation with aluminium(III).

A rapid flow injection (FI) spectrophotometric procedure for tetracycline determination is described. It is based on the injection of a 100 microl sample solution containing tetracycline into merged streams of aluminium(III) chloride (0.01 mol 1(-1)) and Tris-buffer in the presence of KCl (0.06 mol l(-1)), pH 7.0, with the same optimum flow rate of 3.2 ml min(-1). A yellow Al(III)-tetracycline complex was monitored at 376 nm. The flow injection system and the experimental conditions were optimized by means of the univariate method. The procedure was applied to the determination of tetracycline in pharmaceutical preparations with a high sampling rate of at least 165 h(-1). A high precision with a relative standard deviation was obtained less than 0.72 and 0.30% of 5.0 and 10 microg ml(-1) (n=11), respectively. The detection limit (3sigma) and the quantification limit (10sigma) were 0.07 and 0.72 mg l(-1), respectively. There were no interference effects from traditional excipients in the dosage forms when the method was applied to pharmaceutical preparations. The matrix effect could be reduced by the standard addition method.

The development of sequential injection analysis coupled with lab-on-valve for copper determination.

A sequential injection analysis (SIA) using lab-on-valve with air segmentation and spectrophotometric detection was designed for copper(II) determination. It is based on the reaction of copper(II) and 2-carboxy-2'-hydroxy-5'-sulfoformazyl benzene (Zincon) in a weak alkaline solution between the air zones. Beer's Law was obeyed over the range of 0.1-2.0mgL(-1) copper(II) with a correlation coefficient 0.9985 and a slope of 0.2893 absorbance unit/mgL(-1). The relative standard deviation was 2.0% for a series of 10 measurements of 0.5mgL(-1) copper(II) solution. The detection limit (3 S/N) and the limit of quantification (LOQ) were 0.05 and 0.17mgL(-1) respectively. This method has been successfully applied to determination of copper(II) in wastewater with a sample throughput of 120h(-1). The method is superior to the batchwise method in that it provides fully automation, rapidity, less reagents and sample consumption with little waste generation.

Simultaneous micellar LC determination of lidocaine and tolperisone.

A micellar liquid chromatography (MLC) procedure was developed for the simultaneous separation and determination of lidocaine hydrochloride (LD HCl) and tolperisone hydrochloride (TP HCl) using a short-column C18 (12.5 mm x 4.6 mm, 5 microm), sodium dodecyl sulfate (SDS) with a small amount of isopropanol, and diode array detector. The optimum conditions for the simultaneous determination of both drugs were 0.075 mol l(-1) SDS-7.5% (v/v) isopropanol with a flow rate of 0.7 ml min(-1) and detection at 210 nm. The LOD (2S/N) of LD HCl was 0.73 ng 20 microl(-1), whereas that of TP HCl was 1.43 ng 20 microl(-1). The calibration curves for LD HCl and TP HCl were linear over the ranges 0.125-500 microg ml(-1) (r(2)=0.9999) and 1.00-500 microg ml(-1) (r(2)=0.9997), respectively. The %recoveries of both drugs were in the range 98-103% and the %RSD values were less than 2. The proposed method has been successfully applied to the simultaneous determination of TP HCl and LD HCl in various pharmaceutical preparations.