Comparison of high performance liquid chromatography and three titrimetric methods for the determination of ceftazidime in pharmaceutical formulations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Enantioselective determination of ondansetron and 8-hydroxyondansetron in human plasma from recovered surgery patients by liquid chromatography-tandem mass spectrometry

A liquid chromatographic-mass spectrometric assay with atmospheric pressure chemical ionization for quantification of ondansetron and its main metabolite 8-hydroxyondansetron in human plasma was presented. The enantiomeric separation was achieved on a Chiralcel OD-R column containing cellulose tris-(3,5-dimethylphenylcarbamate). The validation data were within the required limits. The assay was successfully applied to authentic plasma samples. Quantitative results from postoperative patients receiving ondansetron demonstrated a great interindividual variability in postoperative plasma drug concentrations, the metabolites were not detected in their unconjugated form. A wide variation in the S-(+)-/R-(-)-ondansetron concentration ratio between 0.14 and 7.18 is indicative for a stereoselective disposition or metabolism. In further studies CYP2D6 and CYP3A4 genotype dependent metabolism of ondansetron enantiomers as well as of co-administered drugs and clinical efficacy of the medication should be tested.

Determination of sexual hormones in liquid cosmetics by polymer monolith microextraction coupled with high performance liquid chromatography

A method was presented for the determination of testosterone, methyltestosterone and progesterone in liquid cosmetics by coupling polymer monolith microextraction (PMME) to high performance liquid chromatography with UV detection. A poly (methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column was selected as the extraction medium, which showed high extraction capacity towards these compounds. To achieve optimum extraction performance, several parameters relating to PMME were investigated, including extraction flow rate and pH value, inorganic salt and organic phase concentration of the sample matrix. By simple dilution with phosphate solution and filtering, the sample solution then could be directly injected into the device for extraction. The limits of detection of testosterone, methyltestosterone and progesterone were calculated to be 2, 3, 2, 8 and 4.6 mu g/L. Good linearity was achieved in the range of 10 to 1000 mu g/L with a linear coefficient. r value above 0. 996. Excellent method reproducibility was found by intra- and inter-day precisions, yielding the relative standard deviations of < 7. 7 % and < 7. 5 %, respectively. Recovery for them in the real samples was between 83% and 119%.

Determination of trace level microcystins in water using solid-phase extraction and high performance liquid chromatography

This paper presents a method for trace level analysis of microcystins in water using solid-phase extraction and high performance liquid chromatography. The optimized condition enabled the determination of common microcystins at levels as low as 0.02 similar to 0.05 mug/L, and the liner range is from 0.1 mug/L to 50 mug/L. The method has been applied to the analysis of field sample from Dianchi lake.

Binding of metal ions with protein studied by a combined technique of microdialysis with liquid chromatography

A method has been developed for the determination of interactions of metal ions and protein by using microdialysis sampling technique combined with pre-column derivation and reversed-phase ion-pair liquid chromatographic (HPLC analysis. Cu(II), Zn(II) and human serum albumin (HSA) were chosen as model metal ions and protein, respectively. The mixed solutions of metal ions and HSA with different molar ratios buffered with 0.1 M Tris-HCl containing 0.1 M NaCl at pH 7.43 were sampled with a mirodialysis probe by keeping perfusion rate at 1 mul/min and the temperature at 37 degreesC. The free concentrations of metal ions in microdialysates were assayed by precolumn derivatization with meso-tetra(4-sulfophenyl)-porphyrin (TPPS4) followed ion-pair HPLC analysis. The recovery (R) of microdialysis sampling was measured in vitro under similar conditions as 65.74% for Cu(II), 70.45% for Zn(II) with R.S.D. below 3.2%. The primary binding constants and number of binding site estimated by the Scatchard plot analysis are 5.04 x 10(6) M-1 and 0.85 for Cu(II), and 9.87 x 10(6) M-1 and 1.10 for Zn(II), respectively. The competition of Cu(II) and Zn(II) at the second binding site on HSA was investigated, and it was observed that there is a second site on HSA to bind Cu(II) and Zn(II), the affinity of Cu(II) is stronger than that of Zn(II) to this second site of HSA. (C) 2001 Elsevier Science B.V. All rights reserved.

Determination of 30 free fatty acids in two famous Tibetan medicines by HPLC with fluorescence detection and mass spectrometric identification

A simple and sensitive high-performance liquid chromatographic (HPLC) method with fluorescence detection and mass spectrometric identification has been developed for analysis of 30 long-chain and short-chain free Fatty acids (FFAs). The fatty acids were derivatized to their esters with 1-[2-(p-toluenesulfonate)ethyl]-2-phenylimidazole-[4,5-f]-9,10-phenanthrene (TSPP) in N,N-dimethylformamide (DMF) at 90 degrees C with anhydrous K2CO3 as catalyst. A mixture Of C-1-C-30 fatty acids was completely separated within 60 min by gradient elution on a reversed-phase C-8 column. Qualitative identification of the acids was performed by atmospheric-pressure chemical ionization mass spectrometry (APCI-MS) in positive-ion mode. The fluorescence excitation and emission wavelengths were 260 and 380 nm, respectively. Quantitative determination of the 30 acids in two Tibetan medicines Gentiana straminea and G. dahurica was performed. The results indicated that the medicines contained many FFAs. Linear correlation coefficients for the FFA derivatives were > 0.9991. Relative standard deviations (RSDs, n = 6) for the fatty acid derivatives were < 3%. Detection limits (at a signal-to-noise ratio of 3:1) were 3.1-38 fmol. When the fatty acid derivatives were determined in the two real samples results were satisfactory and the sensitivity and reproducibility of the method were good.

Determination of free formaldehyde in foundry resins as its 2,4-dinitrophenylhydrazone by liquid chromatography

Formaldehyde is a toxic component that is present in foundry resins. Its quantification is important to the characterisation of the resin (kind and degradation) as well as for the evaluation of free contaminants present in wastes generated by the foundry industry. The complexity of the matrices considered suggests the need for separative techniques. The method developed for the identification and quantification of formaldehyde in foundry resins is based on the determination of free carbonyl compounds by derivatization with 2,4-dinitrophenylhydrazine (DNPH), being adapted to the considered matrices using liquid chromatography (LC) with UV detection. Formaldehyde determinations in several foundry resins gave precise results. Mean recovery and R.S.D. were, respectively, >95 and 5%. Analyses by the hydroxylamine reference method gave comparable results. Results showed that hydroxylamine reference method is applicable just for a specific kind of resin, while the developed method has good performance for all studied resins.

Determination of lipophilic and hydrophilic antioxidants in Solanum Lycopersicum L. tomato from Gordal variety by LL-USAE combined with UHPLC-PDA/FLR

Tomato (Lycopersicon esculentum L.) is one of the main constituents of the Mediterranean diet. Its consumption has been proposed to reduce the risk of cardiovascular diseases and certain types of cancer. It is therefore one of the most popular and extensively consumed vegetable crop worldwide. To gain insights on the potential of Lycopersicon esculentum L. as bioactive food, two analytical methodologies were developed to determine the levels of the lipophilic -tocopherol, α-tocopherol, β-carotene, lycopene; and hydrophilic antioxidants ascorbic acid. The quantification of total carotenoids (β-carotene and lycopene) was assessed through a liquid–liquid ultrasound assisted extraction (LL-USAE) in combination with ultraviolet-visible spectroscopy (UV-Vis), according to method of mean, for total carotenoids (λmáx = 450 nm. The ultra-high performance liquid chromatographic using both photodiode array and fluorescence detection (UHPLC-PDA/FLR), allows the identification and quantification of the target lipophilic and hydrophilic antioxidants. This methodology UHPLC-PDA/FLR is fast, simple and revealed a high sensitivity for the compounds under study. The limits of detection (LODs) and quantification (LOQs) obtained were much lower (about 10 times) than the reported in literature. The method LL-USAE/UV-Vis was validated and applied to different tomato foodstuffs. The results reveal a small increase of carotenoids content during maturation, reaching the maximum level when ripe. These results complement those obtained by the ORAC and TBARS assays that show an increase of antioxidant capacity during maturation. The LODs ans LOQs obtained were also about 10 times lower than reported in literature. The carotenoid content was also evaluated by LL-USAE/UV-Vis in different tomatoes varieties. Regional variety present the high carotenoid level, followed by campari and gordal, and at last grape. This methodology was also applied to different processed food samples containing tomatoes derivatives. Highest carotenoids content were obtained in concentrated tomato foodstuffs.

Development and validation of a new and rapid HPLC for determination of lyophilized teicoplanin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Method development for the simultaneous determination of carboxylic acids, phenolic compounds, and sorbic acid in white wines

A reversed phase liquid chromatographic method was developed for the simultaneous determination of carboxylic acids and phenolics in white wines. The samples, diluted, were injected onto a Spherisorb ODS-2 column with a gradient of sulfuric acid (pH 2.5)/methanol as mobile phase. A diode array detector was used which was set at 210nm for carboxylic acids and altered to 278nm, during the run, far phenolics and sorbic acid. The identification of compounds was based on retention time, co-chromatography and UV spectrum. Some clean-up methods (sep-pak C-18 and an ion exchange column) mere tested and did not improve the results.The analysis was simple, with no sample preparation. Application of this method was illustrated by analyses of Brazilian Welchriesling wines.

Development and Validation of A Stability-Indicating Mekc Method for Determination of Flucloxacillin Sodium in Capsules

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Development and validation of a simple, rapid and stability-indicating high performance liquid chromatography method for quantification of norfloxacin in a pharmaceutical product

A stability-indication high performance liquid chromatographic method has been developed for the determination of norfloxacin in tablet dosage forms. Optimum separation was achieved in less than 7 minutes using Eclipse Plus Zorbax C18 Agilent, 150 mm×4.6 mm i.d., 5 μm particle size column. The analyte was resolved by using a mobile phase 5% acetic acid aqueous solution and methanol (80:20, v/v) at a flow rate 1.0 ml/min on an isocratic high performance liquid chromatographic system at a wavelength of 277 nm. Linearity, system suitability, precision, sensitivity, selectivity, specific, and robustness were established by International Conference Harmonization guidelines. For stress studies the drug was subjected to photolysis, oxidation, acid, alkaline and neutral conditions. The analytical conditions and the solvent developed provided good resolution within a short analysis time and economic advantages. The proposed method not required sophisticated and expensive instrumentation.

Validation of a stability-indicating HPLC method with diode array detection for the determination of beclomethasone dipropionate in aqueous suspension for nebulizer

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Determination of appropriate condition on replacing methane from hydrate with carbon dioxide

This paper is intended to determine the appropriate conditions for replacing CH4 from NGH with CO2. By analyzing the hydration equilibrium graphs and geotherms, the HSZs of NGH and CO2 hydrate, both in permafrost and under deep sea, were determined. Based on the above analysis and experimental results, it is found that to replace CH4 from NGH with gaseous CO2, the appropriate experimental condition should be in the area surrounded by four curves: the geotherm, (H-V)(CO2), (L-V)(CO2) and (H-V)(CH4), and to replace CH4 from NGH with liquid CO2, the condition should be in the area surrounded by three curves: (L-V)(CO2), (H-L)(CO2) and (H-V)CH4. For conditions in other areas, either CO2 can not form a hydrate or CH4 can release little from its hydrate, which are not desirable results.

Prediction of soil organic carbon partition coefficients by soil column liquid chromatography

To avoid the limitation of the widely used prediction methods of soil organic carbon partition coefficients (K-OC) from hydrophobic parameters, e.g., the n-octanol/water partition coefficients (K-OW) and the reversed phase high performance liquid chromatographic (RP-HPLC) retention factors, the soil column liquid chromatographic (SCLC) method was developed for K-OC prediction. The real soils were used as the packing materials of RP-HPLC columns, and the correlations between the retention factors of organic compounds on soil columns (k(soil)) and K-OC measured by batch equilibrium method were studied. Good correlations were achieved between k(soil) and K-OC for three types of soils with different properties. All the square of the correlation coefficients (R-2) of the linear regression between log k(soi) and log K-OC were higher than 0.89 with standard deviations of less than 0.21. In addition, the prediction of K-OC from K-OW and the RP-HPLC retention factors on cyanopropyl (CN) stationary phase (k(CN)) was comparatively evaluated for the three types of soils. The results show that the prediction of K-OC from k(CN) and K-OW is only applicable to some specific types of soils. The results obtained in the present study proved that the SCLC method is appropriate for the K-OC prediction for different types of soils, however the applicability of using hydrophobic parameters to predict K-OC largely depends on the properties of soil concerned. (C) 2004 Elsevier B.V. All rights reserved.