A sensitive and selective high performance liquid chromatography-peroxyoxalate chemiluminescence (PO-CL) method has been developed for the simultaneous determination of chlorpheniramine (CPA) and monodesmethyl chlorpheniramine (MDCPA) in human serum. The method combines fluorescent labeling with 4-(4,5-diphenyl-1H-imidazole-2-yl)phenyl boronic acid using Suzuki coupling reaction with PO-CL detection. CPA and MDCPA were extracted from human serum by liquid-liquid extraction with n-hexane. Excess labeling reagent, which interfered with trace level determination of analytes, was removed by solid-phase extraction using a C18 cartridge. Separation of derivatives of both analytes was achieved isocratically on a silica column with a mixture of acetonitrile and 60 mM imidazole-HNO(3) buffer (pH 7.2; 85:15, v/v) containing 0.015% triethylamine. The proposed method exhibited a good linearity with a correlation coefficient of 0.999 for CPA and MDCPA within the concentration range of 0.5-100 ng/mL. The limits of detection (S/N = 3) were 0.14 and 0.16 ng/mL for CPA and MDCPA, respectively. Using the proposed method, CPA could be selectively determined in human serum after oral administration.

A simple chemiluminometric method using flow injection has been developed for the determination of paracetamol (acetaminophen), based on the chemiluminescence produced by the reduction of tris(2,2'-bipyridyl)ruthenium(III). The latter is obtained by oxidation of tris(2,2'-bipyridyl)ruthenium(II) by potassium permanganate in dilute sulphuric acid in the presence of paracetamol. A standard or sample solution was injected into the ruthenium(II) stream (flow rate 1.5mlmin(-1)) which was then merged with potassium permanganate in dilute sulphuric acid stream (flow rate 0.5mlmin(-1)). The chemiluminescence intensity is enhanced by the presence of manganese(II) ions. Under the optimum conditions, a linear calibration graph was obtained over the range of 0.3-50.0mugml(-1) and the detection limit was 0.2mugml(-1)(s/n=3). The relative standard deviation of the proposed method calculated from 20 replicate injections of 5.0mugml(-1) paracetamol was 1.1%. The sample throughput was 90h(-1). The method was successfully applied to the determination of paracetamol in commercial pharmaceutical formulations.

A new flow-injection procedure for the determination of phenylpropanolamine and methoxamine is proposed. The method is based on the derivatization reaction of the primary amine group with o-phthalaldehyde in the presence of 2-mercaptoethanol using fluorimetric detection. The calibration graphs based on peak areas were linear in the ranges 5-200 ng ml(-1) for phenylpropanolamine and 0.2-6 ng ml(-1) for methoxamine. The detection limits were 3.8 and 0.13 ng ml(-1), respectively. The methods were applied to the determination of the drugs in commercial pharmaceutical preparations.

A simple and reliable high-performance liquid chromatographic method was developed for the simultaneous determination of mixture of phenylephrine hydrochloride (PHENYL), guaifenesin (GUAIF), and chlorpheniramine maleate (CHLO) either in pure form or in the presence of methylparaben and propylparaben in a commercial cough syrup dosage form. Separation was achieved on a C8 column using 0.005 M heptane sulfonic acid sodium salt (pH 3.4 +/- 0.1) and acetonitrile as a mobile phase by gradient elution at different flow rates, and detection was done spectrophotometrically at 210 nm. A linear relationship in the range of 30-180, 120-1800, and 10-60 microg/mL was obtained for PHENYL, GUAIF, and CHLO, respectively. The results were statistically analyzed and compared with those obtained by applying the British Pharmacopoeia (2002) method and showed that the proposed method is precise, accurate, and can be easily applied for the determination of the drugs under investigation in pure form and in cough syrup formulations.

Multivariate spectrophotometric calibration and liquid chromatography (LC) methods were used for the simultaneous determination of the active ingredients in 2 multicomponent mixtures containing chlorpheniramine maleate and phenylpropanolamine hydrochloride with ibuprofen and caffeine (mixture 1) or with propyphenazone (mixture 2). For the multivariate spectrophotometric calibration methods, principal component regression (PCR) and partial least squares (PLS-1), a calibration set of the mixtures consisting of the components of each mixture was prepared in distilled water. A leave-1-out cross-validation procedure was used to find the optimum numbers of latent variables. Analytical parameters such as sensitivity, selectivity, analytical sensitivity, limit of quantitation, and limit of detection were determined for both PLS-1 and PCR. The LC method depends on the use of a cyanopropyl column with the mobile phase acetonitrile-12 mM ammonium acetate, pH 5.0 (25 + 75, v/v), for mixture 1 or acetonitrile-10 mM potassium dihydrogen phosphate, pH 4.7 (45 + 55, v/v), for mixture 2; the UV detector was set at 212 nm. In spite of the presence of a high degree of spectral overlap of these components, they were rapidly and simultaneously determined with high accuracy and precision, with no interference from the matrix excipients. The proposed methods were successfully applied to the analysis of pharmaceutical formulations and laboratory-prepared mixtures containing the 2 multicomponent combinations.

Three methods are developed for the determination of two multicomponent mixtures containing guaiphenesine (GU) with salbutamol sulfate (SL), methylparaben (MP) and propylparaben (PP)[mixture 1]; and acephylline piperazine (AC) with bromhexine hydrochloride (BX), methylparaben (MP) and propylparaben (PP)[mixture 2]. The resolution of the two multicomponent mixtures has been accomplished by using numerical spectrophotometric methods such as partial least squares (PLS-1) and principal component regression (PCR) applied to UV absorption spectra of the two mixtures. In addition HPLC method was developed using a RP 18 column at ambient temperature with mobile phase consisting of acetonitrile-0.05 M potassium dihydrogen phosphate, pH 4.3 (60:40, v/v), with UV detection at 243 nm for mixture 1, and mobile phase consisting of acetonitrile-0.05 M potassium dihydrogen phosphate, pH 3 (50:50, v/v), with UV detection at 245 nm for mixture 2. The methods were validated in terms of accuracy, specificity, precision and linearity in the range of 20-60 microg ml(-1) for GU, 1-3 microg ml(-1) for SL, 20-80 microg ml(-1) for AC, 0.2-1.8 microgml(-1) for PP and 1-5 microg ml(-1) for BX and MP. The proposed methods were successfully applied for the determination of the two multicomponent combinations in laboratory prepared mixtures and commercial syrups.

Paracetamol is a common analgesic and antipyretic drug that is used for the relief of fever, headaches and other minor aches and pains. Their determination in pharmaceuticals is of paramount importance, since an overdose of paracetamol can cause fulminating hepatic necrosis and other toxic effects. Many analytical methodologies have been proposed for the determination of paracetamol. The aim of the present study is to evaluate the utility of different techniques for quantification of paracetamol content in pharmaceutical formulations and biological samples.

An accurate, simple, reproducible, and sensible liquid chromatographic method was developed and validated for the determination of chlorpheniramine maleate and dexamethasone in a tablet formulation. The analysis was performed at room temperature on a reversed-phase C18 column with UV detection at 254 nm. The mobile phase consisted of 7.5 mM monobasic potassium phosphate in methanol-water (62.5 + 37.5) at a constant flow rate of 1 mL/min. The method was validated in terms of linearity, precision, accuracy, and specificity by forced decomposition of chlorpheniramine maleate and dexamethasone initiated by using acid, base, water, hydrogen peroxide, heat, and light. The response was linear in the ranges of 0.04-0.12 and 0.006-0.016 mg/mL for chlorpheniramine maleate (r2 = 0.9999) and dexamethasone (r2 = 0.9994), respectively. The relative standard deviation values for intra- and interday precision studies were 2.39 and 2.02, respectively, for chlorpheniramine maleate and 2.39 and 1.25, respectively, for dexamethasone. Recoveries ranged from 95.07 to 101.95% for chlorpheniramine maleate and from 97.75 to 102.10% for dexamethasone.

A stability indicating high performance liquid chromatography procedure has been developed for the simultaneous determination of guaifenesin (GUA), methyl p-hydroxybenzoate (MHB) and propyl p-hydroxybenzoate (PHB) in a commercial cough syrup dosage form. The method was specific and stability indicating as chromatographic conditions were selected to provide adequate separation of GUA, MHB and PHB from the putative degradation products guaiacol (GUAI) and p-hydroxybenzoic acid (HBA) as well as from excipients. The isocratic separation and quantitation were achieved within 17min on a 150-mm column with an ether-linked phenyl stationary phase and a hydrophilic endcapping. The mobile phase was constituted of eluant A: aqueous phosphate buffer (pH 3.0, 10mM)/acetonitrile 25/75 (v/v) and eluant B:methanol; the A:B ratio was 85:15 (v/v) with a flow rate 1mlmin(-1) and detection of analytes at 254 and 276nm. The method showed good linearity for the GUA-MHB-PHB mixture in the 95-285, 4-12, and 1-3mugml(-1) ranges, respectively, being all the square of the correlation coefficients greater than 0.999. The interday R.S.D.s were 1.17, 1.14, and 0.91%, for GUA, MHB, and PHP, respectively. The method demonstrated also to be accurate; indeed the average recoveries, at 100% of the target assay concentration, were 100.5, 100.3, and 100.7% with relative standard deviations of 0.8, 0.7, and 0.4% for GUA, MHB, and PHB, respectively, from laboratory prepared samples. The applicability of the method was evaluated in commercial dosage form analysis as well as in stability studies.

A simple and rapid gas chromatographic (GC) method has been developed for the simultaneous determination of combinations of acetaminophen, phenylpropanolamine hydrochloride, guaifenesin, pseudoephedrine hydrochloride, caffeine, chlorpheniramine maleate, and dextromethorphan hydrobromide in cough and cold tablets and syrups. After extraction of the analyte with alkaline ethyl acetate, 2 microL extract was injected (splitting ratio of 50:1) into a gas chromatograph equipped with a CBP1-M25-025 fused silica capillary column (25 m x 0.22 mm; film thickness, 0.25 microm). The column temperature was held at 150 degrees C for 5 min, increased to 175 degrees C at 3 degrees C/min, and increased to 270 degreesC at 10 degrees C/min. The temperatures of the flame ionization detector and injector were maintained at 300 degrees C. The GC method is inexpensive, rapid, accurate, and precise, and thus it can be used for routine analysis of tablet and syrup preparations in quality control laboratories of pharmaceutical companies.

Koi herpesvirus (KHV) is the aetiological agent of an emerging disease (KHVD) associated with mass mortalities in koi and common carp and reported from at least 30 countries. We report the first isolation of KHV from koi and common carp in Indonesia and initial characterization of the isolates. Clinical signs, histopathology and virion morphology are similar to those of isolates from other countries. Phylogenetic analyses using the thymidine kinase gene amplified from each isolate and from carp tissue samples collected from KHVD outbreaks throughout Indonesia indicated that the Indonesian isolates are more closely related to the Asian than the European KHV lineage. Sequence analysis of two other variable regions between ORF29 and ORF31 (marker I) and near the start of ORF 133 (marker II) indicated that all Indonesian isolates displayed a marker I allele (I(++)) previously identified only in isolates of the Asian lineage. However, in the marker II region, all Indonesian isolates displayed the II(-) allele, which has been reported previously only amongst isolates of the European lineage, and nine of these displayed a mixed genotype (II(+)II(-)). The I(++)II(-) genotype has not been reported previously and appears to represent a new intermediate lineage that may have emerged in Indonesia.

In 2002, the suspected koi herpesvirus (KHV) outbreak in Indonesia was investigated by an International Emergency Disease Control Task Force organized by NACA immediately following a request for assistance by the Government of Indonesia. The Task Force gained immediate support from ACIAR, AAHRI, FAO, CENTEX-Thailand, INTERVET, Stirling University, and the University of California. The Task Force findings revealed the involvement of an infectious agent, an analogy with KHV outbreaks, its introduction through fish importation and its spread into other areas through fish movement. A number of actions taken by the Government of Indonesia included temporary restrictions, through a Ministry Circular, on the movement of koi and common carps, intensive information dissemination, emergency notification to the OIE, and early warning to neighbouring countries. FAO immediately responded by providing an emergency technical assistance project to improve Indonesia's national capacity to respond to the carp disease epizootic. Recognizing the significance of and necessity for:(a) enhancing regional and international cooperation;(b) improving awareness of emerging disease epizootics and improving diagnostic capabilities at both national and regional levels;(c) proactive reporting of serious disease outbreaks as a mechanism for early warning;(d) detailed documentation of outbreak scenarios;(e) emergency preparedness;(f) empowering farmers to manage disease and other risks; and (g) strong national commitment from concerned authorities are some of the important lessons learned. Despite the intense efforts, the spread of KHV did not stop and continued to affect other countries such as Japan, China, the Taiwan Province of the Republic of China, and most recently Thailand. All relevant stakeholders cannot afford to be complacent and we need innovative responses to current and future disease challenges. The lessons learned in managing the Indonesian KHV outbreak as well as lessons from past disease epizootics will hopefully assist us in improving preparedness and responses to similar events when they occur in the future.

3-O-[beta-D-glucopyranosyl-(1"--> 2')-beta-D-glucopyranosyl], 27-O-beta-D-glucopyranosyl-(25R)-spirost-5-ene-3beta,27-diol was isolated from cell suspension cultures of Costus speciosus, following incubation with diosgenin, and its structure was elucidated using a combination of one- and two-dimensional 1H and 13C NMR spectral data, and positive and negative ion ESMS spectral data.

A new biotransformation product, salicyl alcohol-7-O-beta-D-(beta-1,6-D-glucopyranosyl)-gluco-pyranoside was isolated from cell suspension cultures of Solanum laciniatum, following administration of salicyl alcohol, and its structure was elucidated using a combination of one- and two-dimensional 1H and 13C-NMR data, and positive and negative ion ESMS data.

A new biotransformation product, salicyl alcohol-7-O-beta-D-(beta-1,6-D-glucopyranosyl)-gluco-pyranoside was isolated from cell suspension cultures of Solanum laciniatum, following administration of salicyl alcohol, and its structure was elucidated using a combination of one- and two-dimensional 1H and 13C-NMR data, and positive and negative ion ESMS data.

Cell suspension cultures of Solanum laciniatum were able to transform exogenously inoculated salicyl alcohol into salicyl alcohol 7-O-beta-D-glucopyranoside (isosalicin). The highest level of isosalicin (54.6 mg/g dry weight) in the cells was formed within 2 days after inoculation with salicyl alcohol (37.5 mg/flask containing 50 ml of medium). The biotransformation capacity of the cell suspension cultures was about 31.1%.

Two new biotransformation products, N-acetyl-m-aminobenzoic acid and N-formyl-m-aminobenzoic acid were isolated from cell suspension cultures of Solanum laciniatum following administration of m-aminobenzoic acid, and their structures were elucidated using one- and two-dimensional 1H- and 13C-NMR data.

A simple and rapid gas chromatographic (GC) method has been developed for the simultaneous determination of combinations of acetaminophen, phenylpropanolamine hydrochloride, guaifenesin, pseudoephedrine hydrochloride, caffeine, chlorpheniramine maleate, and dextromethorphan hydrobromide in cough and cold tablets and syrups. After extraction of the analyte with alkaline ethyl acetate, 2 microL extract was injected (splitting ratio of 50:1) into a gas chromatograph equipped with a CBP1-M25-025 fused silica capillary column (25 m x 0.22 mm; film thickness, 0.25 microm). The column temperature was held at 150 degrees C for 5 min, increased to 175 degrees C at 3 degrees C/min, and increased to 270 degreesC at 10 degrees C/min. The temperatures of the flame ionization detector and injector were maintained at 300 degrees C. The GC method is inexpensive, rapid, accurate, and precise, and thus it can be used for routine analysis of tablet and syrup preparations in quality control laboratories of pharmaceutical companies.

The use of a UV double divisor-ratio spectra derivative calibration for the simultaneous analysis of synthetic samples and commercial tablet preparations without prior separation is proposed. The method was successfully applied to quantify three ternary mixtures, chlorpheniramine maleate and caffeine combined with paracetamol or acetylsalicylic acid and a mixture of acetylsalicylic acid combined with paracetamol and caffeine, using the information in the absorption spectra of appropriate solutions. Beer's law was obeyed in the concentration range of 0.84-4.21 microg/ml for chlorpheniramine maleate, 1.60-15.96 microg/ml for caffeine, 2.0-20.0 microg/ml for acetylsalicylic acid and 1.58-15.93 microg/ml for paracetamol. The whole procedure was applied to synthetic mixtures of pure drugs as well as to commercial preparations (Algon) by using content uniformity and dissolution tests (USP 24) and was found to be precise and reproducible. According to the dissolution profile test more than 84% of paracetamol and caffeine were dissolved within 20 min. Acetylsalicylic acid dissolved more slowly, taking about 45-60 min to dissolve completely. A chemometric method partial least squares (PLS) and a HPLC method were also employed to evaluate the same mixtures. The results of the proposed method were in excellent agreement with those obtained from PLS and HPLC methods and can be satisfactorily used for routine analysis of multicomponent dosage forms.

In this contribution, a new separation method for simultaneous determination of paracetamol, caffeine, acetylsalicylic acid, and internal standard benzoic acid was developed based on a novel reversed-phase sequential injection chromatography (SIC) technique with UV detection. A Chromolith Flash RP-18e, 25-4.6mm column (Merck, Germany) and a FIAlab 3000 system (USA) with an 8-port selection valve and a 5 mL syringe were used for sequential injection chromatographic separations in our study. The mobile phase used was acetonitrile-(0.01 M) phosphate buffer (10:90, v/v) pH 4.05, flow rate 0.6 mL min(-1). UV detection was at 210 and 230 nm. The validation parameters showed good results: linearity (r >0.999) for all compounds, detection limits in the range 0.3-0.8 microg mL(-1), repeatability (RSD) of peak heights between runs in the range 1.10-4.30% at three concentration levels and intra-day repeatability of the retention times in the range 0.28-0.43%. The analysis time was <6 min. The method was found to be applicable for the routine analysis of the active compounds paracetamol, caffeine, and acetylsalicylic acid in pharmaceutical tablets.

The optimization of RP-HPLC method defined the simultaneous influence of some important conditions, such as the mobile phase composition, pH of the mobile phase and temperature, on the separation and determination. The RP-HPLC method was done for the determination of paracetamol, caffeine and propyphenazone in a multicomponent pharmaceutical dosage form. The separation factor values define the optimal conditions, which were confirmed by analysing the appropriate mathematical models. The chromatographic system Hewlett Packard 1100 consisted of a HP 1100 pump, HP 1100 UV-VIS detector and HP integrator. Separations were performed on a Beckman Ultrasphere ODS 4.6 x 150 mm, 5 microns particle size column. Samples were introduced through a Rheodyne injector valve with a 20 microL sample loop. UV detection was performed at 265 nm and phenobarottons was used as an internal standard. The optimization was performed within the pH range from 2.5 to 6.0; temperature range from 20 degrees C to 55 degrees C and composition of the mobile phase methanol-water from (30:70 V/V) to (65:35 V/V). The three-D graphs, constructed with sixty-four experimental points, confirmed the optimal conditions for the determination of the investigated analgetic drugs.

A micellar electrokinetic chromatography (MEKC) method was established for determination of paracetamol (PARA) and chlorpheniramine maleate (CPM) in cold tablets. Separation of both drugs, as well as other seven cold remedy ingredients, was achieved in 25.5 min using a sodium dihydrogenphosphate-sodium tetraborate buffer (10 mM, pH 9.0) containing sodium dodecyl sulfate (SDS)(50 mM) and acetonitrile (26% v/v). The effective capillary length of 50 cm, the separating voltage of 15 kV and the temperature of 30 degrees C was optimized. Detection was by a diode array detector at 214 nm. Method linearity was excellent (r(2)>0.999) over the concentration tested (10-250 microg/ml) with good precision and accuracy. Recoveries were good (>99%) with limits of detection of 0.4 and 0.5 microg/ml and limits of quantitation of 2 (%R.S.D.=3.1%) and 4 (%R.S.D.=2.4%) microg/ml, for PARA and CPM, respectively. The developed method was applied to the determination of ingredients in cold tablets and was found to be simple, rapid and efficient.

A procedure was developed and validated for measuring chlorpheniramine maleate and tincture ipecac (as emetine hydrochloride) by reversed-phase liquid chromatography with methanol-10 mM sodium heptanesulfonate (20 + 30) as the mobile phase; the pH was adjusted to 4 with acetic acid, and the flow rate was at 1.5 mL/min, with ultraviolet detection at 254 nm. Propyl paraben was used as the internal standard. The standard curves were linear (r = 0.998 and 0.9998) for both chlorpheniramine maleate and emetine hydrochloride over the ranges of 5-100 and 0.1-40 microg/mL, respectively. The mean recoveries +/- standard deviation were 101.37 +/- 2.77% for chlorpheniramine maleate and 98.8 +/- 1.47% for emetine hydrochloride. The proposed method was applied to the determination of chlorpheniramine maleate alone in tablet and syrup dosage forms. The method also was applied to the determination of the emetine content of ipecac liquid extract and tincture ipecac; the results were compared with those of the method of the British Pharmacopoeia. The proposed method was applied successfully to the simultaneous determination of chlorpheniramine maleate and tincture ipecac, as emetine hydrochloride, in syrup dosage form. Both drugs and the internal standard were separated from all interfering components in < 5 min. The proposed method is simple, specific, and economical, when compared with other published methods that determine each component alone.

Compound amantadine tablets is a new drug for virus-common cold. It was consisted of paracetamol, amantadine hydrochloride and caffeine etc. Because of the disturbing of the caffeine, the paracetamol can not be determined by UV directly. So the paracetamol in compound amantadine tablets was determined by derivative spectrophotometry. This method is simple and rapid, no distillation and no separation, with average recovery of 100.0%, and RSD of 0.36%.

Principal component regression (PCR) method is used to analyse five components: acetaminophen, p-aminophenol, caffeine, chlorphenamine maleate and guaifenesin. The basic principle and the analytical step of the approach are described in detail. The computer program of LHG is based on VB language. The experimental result shows that the PCR method has no systematical error as compared to classical method. The experimental result shows that the average recovery of each component is all in the range from 96.43% to 107.14%. Each component obtains satisfactory result without any pre-separation. The approach is simple, rapid and suitable for the computer-aid analysis.

An HPLC method is described for the determination of chlorpheniramine, paracetamol and pseudoephedrine hydrochloride simultaneously in Nuonuo composite tablets. Chromatographic conditions were: column, YWG-C18 column (150 mm x 4.0 mm i.d., 10 microns); mobile phase, 25% methanol(0.05 mol/L sodium dihydrogen phosphate); flow rate, 1.0 mL/min; detection wavelength, 205 nm; injection volume, 20 microL. The detection limits of chlorpheniramine, paracetamol and pseudo-ephedrine hydrochloride were 1.16 mg/L, 0.15 mg/L and 1.82 mg/L respectively and the corresponding average recoveries were 98.35%(n = 5, RSD = 1.60%), 101.16%(n = 5, RSD = 1.50%) and 98.50%(n = 5, RSD = 1.59%) respectively. The proposed method is simple, rapid and accurate and has been satisfactorily used for the quantitative analysis of real samples.