Evaluation of antioxidant activity of hydrophilic and lipophilic compounds in edible oils by a novel fluorimetric method.

A novel fluorimetric method is described for the evaluation of the antioxidant activity of hydrophilic and lipophilic compounds and complex natural products such as edible oils. The method is based on the measurement of fluorescence emission intensity of N-methylacridone produced during the reaction of lucigenin with hydrogen peroxide. The presence of antioxidants in the sample inhibits the concentration of N-methylacridone and reduces the fluorescence intensity. The method was fully validated and applied to a variety of hydrophilic and lipophilic compounds as well as to various types of edible oils and their corresponding hydrophilic and lipophilic extracts. Results were compared to those derived from a lucigenin based chemiluminescent method and the Folin-Ciocalteau method for total phenols. The differences in total antioxidant activity of edible oils of various origins and the effect of heating on total antioxidant activity was further studied and discussed.

Development and validation of a chemiluminogenic method for the evaluation of antioxidant activity of hydrophilic and hydrophobic antioxidants.

A sensitive and simple procedure is described for evaluating the antioxidant activity of 21 known hydrophilic and hydrophobic compounds by using the chemiluminogenic reaction of lucigenin with hydrogen peroxide. The method was validated for linearity, limit of detection, precision, additivity and ruggedness and compared to the DPPH method. Furthermore, the IC(50) values of all compounds were calculated and compared by both methods and results are discussed on the basis of structure-antioxidant activity relationship. Among hydrophilic antioxidants, phenolic compounds with adjacent hydroxyl groups (gallic acid, caffeic acid, pyrocatechol, (+/-)-catechin, oleuropein) were found to be efficient antioxidants in chemiluminescence method with IC(50) values at the sub-micromolar range, while phenolic compounds without adjacent hydroxyl groups (p-coumaric acid, BHT, BHA, resorcinol) exhibited weak activity with IC(50) values>10microM. Alpha-tocopherol was the most efficient hydrophobic antioxidant (IC(50)=6.7microM) while oleic and linoleic acids were found to be very poor antioxidants (IC(50)>300microM). Finally the method was applied to the estimation of antioxidant activity of complex samples such as olive oils. To the best of our knowledge, this is the first time that total antioxidant activity can be determined directly in olive oil and not in aqueous extracts.

Two-step silylation procedure for the unified analysis of 190 doping control substances in human urine samples by GC-MS.

BACKGROUND: While a number of different derivatization procedures for screening GC-MS analysis of prohibited substances are followed by doping control laboratories, a unified derivatization procedure for the GC-MS analysis of 190 different doping agents was developed. RESULTS: Following preliminary experiments, a two-step derivatization procedure was selected. The evaluation of various silylation parameters, such as reagent composition, reaction time, reaction temperature, catalysts and microwave oven reaction time, for this procedure was carried out. CONCLUSION: The suitability of the developed procedure was demonstrated through application on urine samples at concentration levels of the minimum required performance limit for all tested substances. This new derivatization procedure, which significantly decreases time and cost, is suitable for a routine basis application.

Analytical applications of flow injection with chemiluminescence detection--a review.

This paper reviews the literature on analytical applications of flow injection (FI) techniques with chemiluminescence (CL) detection from 1995-1999. The focus is on the application of FI-CL to the quantitative determination of specific analytes in real sample matrices. Therefore, entries have been tabulated under the most appropriate application area, ie pharmaceutical, environmental, foods and beverages and biomedical, as defined by the matrix that has been analysed. Each table lists analytes alphabetically and gives details of the exact sample matrix, the limit of detection (as reported in the original paper) and comments on the CL reaction used.

Chemiluminometric determination of reserpine and related alkaloids.

The determination of the alkaloids reserpine, rescinnamine and yohimbine based on a chemiluminogenic reaction with potassium permanganate in the presence of polyphosphoric acid is described. The investigation was carried out using a batch and a flow injection chemiluminometer. Both approaches were accurate and precise, allowing the measurement of reserpine within the ranges 0.100-3.00 and 0.050-3.00 micrograms ml-1 with RSD values for 1.00 microgram ml-1 of 1.91 and 0.33% (n = 8) with the batch and the flow injection manifold, respectively. The procedure was successfully applied to formulations after extraction of reserpine with chloroform, with recoveries from commercial formulations within the range 95.2-99.0%.

Detection in the liquid phase applying chemiluminescence.

Several chemiluminescence-based reactions are applicable to the determination of various bio-pharmaceutically important analytes, and they can be applied for monitoring chemiluminescence emission using flow injection, liquid chromatographic and capillary electrophoretic analysis, as well as for the development of chemiluminescence-based sensors or in immunoassays. As in general the emission intensity is linearly proportional to the concentration of any of the reagents, the technique allows the analysis of different species involved in the light-producing reaction, amongst which are the chemiluminescent reagent, oxidants, inhibitors, cofactors, catalysts, some fluorophore, etc. The present overview illustrates some important applications of the last decade on this rather unfamiliar luminescence technique to detectional challenges in the liquid phase. The required instrumentation is limited as no external light source is needed. Also, the technique opens perspectives for increasing detection sensitivity in miniaturized flowing streams. On the other hand, several drawbacks still limit full application, eg dependence of the emission signal upon a number of environmental factors forcing the analyst to make a compromise between separating and measuring conditions, a lack of selectivity in specific cases, the critical detection of the signal at strictly defined periods, especially in the case of sharp emission vs time profiles, and the development of detection devices in capillary electrophoresis.

Chemiluminescence-based detection: principles and analytical applications in flowing streams and in immunoassays.

The present paper provides the principles of chemiluminescence (CL) and its powerful applications in analytical chemistry, mainly in the area of flow injection analysis, column liquid chromatographic and capillary electrophoretic separating systems, and its potential in immunoassays. CL is light produced by a chemical reaction. The most common advantages of chemiluminescent reactions are the relatively simple instrumentation required, the very low detection limits and wide dynamic ranges, which have contributed to the interest of CL detection in flow injection analysis, high performance liquid chromatography, including miniaturized systems, and, most recently, the exploding area of capillary electrophoresis. The latter powerful microanalytical separation technique offers high numbers of theoretical plates and relatively short analysis times requiring only small sample volumes, the migrating system comprising aqueous buffer solutions. In non-isotopic immunoassays, covering a great variety of applications in human and veterinary medicine, forensic medicine, agriculture and food industry, the radioisotope is replaced by a fluorescence or chemiluminescent label. The use of CL as a detection principle permits quantitative determination of various compounds at low concentrations. Disadvantages of the CL-based technique may include lack of sufficient selectivity and sensitivity to various physicochemical factors.

Cerium (IV)-based chemiluminescence analysis of hydrochlorothiazide.

A flow-injection analytical method for the determination of hydrochlorothiazide is presented. The method is based on the chemiluminescence reaction of hydrochlorothiazide with cerium(IV) in sulphuric acid, sensitized by the fluorescent dye rhodamine 6G. The proposed procedure allows quantitation of hydrochlorothiazide in the concentration range of 0.33-130 mumol l(-1) with a detection limit of 0.15 mumol l(-1), an RSD of 2.4% at 10 mumol l(-1) and a sample measurement frequency of 200 h(-1). The method was successfully applied to the determination of hydrochlorothiazide in pharmaceutical preparations containing, amongst others, lactose, maize starch, calcium phosphate, magnesium stearate, potassium chloride and E 110 (disodium-6-hydroxy-5-(4-sulphonatophenylazo) naphthalene-2-sulphonate) as the concomitant species. Apart from the single formulation, hydrochlorothiazide was also determined in tablets combined with the antihypertensive lisinopril.

Chemiluminescence determination of tiopronin by flow injection analysis based on cerium(IV) oxidation sensitized by quinine.

A flow injection analysis method is proposed for the determination of tiopronin based upon the oxidation by cerium(IV) in dilute sulfuric acid medium and sensitized by quinine. With the peak height as a quantitative parameter applying optimum working conditions, tiopronin is determined over the 1-400 microM range (150 microliters per injection, n = 10, r = 0.9994) with a detection limit of 0.34 microM and an RSD (n = 10) less than 2% at 20 and 50 microM. The proposed method, combining the advantages of speed and sensitivity, was applied to the routine determination of tiopronin in a pharmaceutical preparation.

Chemiluminescence in drug assay.

The phenomenon of chemiluminescence allows the development of simple analytical procedures using low cost instrumentation which can be easily controlled by computers. The sensitivity of the methods is excellent, and selectivity is improved by high-performance liquid chromatographic separation. A broad area of chemiluminometric applications is drug analysis, which is reviewed in this paper.

Chemiluminescence analysis of captopril: comparison between luminol and rhodamine B-sensitized cerium(IV) methods.

Two chemiluminescence methods for the determination of captopril are compared in the present paper. The first method is based on the reaction of captopril with the luminol-hydrogen peroxide-copper(II) system, the copper(II) ion acting as a key species in the light-emitting process. The catalytic activity of copper(II) decreases due to complex formation with the sulphhydrylic captopril analyte. Application of this indirect method allows captopril determinations in the 8.0-1.0 microM range as starting concentrations. Similarly, captopril was capable of generating chemiluminescence from acid cerium(IV) solutions, a reaction that could be analytically exploited by the inclusion of the rhodamine B fluorophore yielding a sensitized-type of chemiluminescence emission that allowed quantitation of captopril concentrations in the 0.1-6.0 microM range with a detection limit of 0.037 microM (original concentration). For both types of reaction the experimental conditions were optimized and a direct application was carried out on a commercial drug formulation.

Flow injection chemiluminometric determination of epinephrine, norepinephrine, dopamine and L-dopa.

A method is proposed for the determination of 0.0500-1.00 microgram ml-1 of epinephrine and L-dopa and 0.100-1.00 micrograms ml-1 of norepinephrine and dopamine by their chemiluminogenic oxidation with potassium permanganate in acidic medium, in the presence of formaldehyde, which greatly improves the sensitivity. Flow injection allows the measurement of 80 solutions per hour. The method was also optimized for a continuous-flow system. Comparative results from numerous organic compounds proved the necessity for electron-donating groups on the benzene ring for sensitive chemiluminescent characteristics.

Continuous-flow chemiluminometric determination of tetracyclines in pharmaceutical preparations and honey by oxidation with N-bromosuccinimide.

A continuous flow chemiluminometric method for determining 0.050-3.00 micrograms cm-3 of tetracycline, 0.50-5.00 micrograms cm-3 of oxytetracycline, 0.50-7.00 micrograms cm-3 of doxycycline and chlorotetracycline and 0.30-3.00 micrograms cm-3 of demeclocycline in pharmaceutical preparations and honey is described. The method is based on the chemiluminescence produced by the action of N-bromosuccinimide on tetracyclines in alkaline solution. The emission intensity is greatly enhanced by the presence of ammonia. The procedure is automated and solutions can be analysed at a rate of 130 h-1 with a relative error of about 2%. During evaluation of possible interferences of the method, recoveries from solutions with common excipients and other concomitant compounds were in the ranges 97.8-104.0 and 96.6-109.4%, respectively. Recoveries of various tetracyclines from commerical formulations and honey samples were in the ranges 95.2-103.7 and 89.2-106.6%, respectively. The results obtained for the assay of commerical pharmaceutical preparations compared well with those obtained by an official method and demonstrated accuracy and precision (< or = 5%).