Spectrophotometric determination of Cu(II) with sequential injection analysis

A sensitive method is presented for the determination of the synthetic antioxidant butylated hydroxyanisole (BHA) based on the dansylation process of the phenolic hydroxy group. The fluorescence developed can be measured directly without previous extraction or chromatographic separation of the labelled fluorescent compound. It is shown the effect of numerous experimental variables affecting the fluorescence intensity and the signal-to-noise ratio of the dansyl derivative. The compound was determined over the range 0.05-5 mug ml(-1), with a relative S.D. of 3.8% (300 ng ml(-1)) and a detection limit of 52 ng ml(-1). The selectivity conferred by the dansylation reaction has permitted to avoid the interference of normally accompanying antioxidants, such as butylated hydroxytoluene (BHT), due to steric impediment. The stability of the DNS-derivative is well suited for the analysis of different foodstuffs.     

Simultaneous determination of the camptothecin analogue CPT-11 and its active metabolite SN-38 by high-performance liquid chromatography: application to plasma pharmacokinetic studies in cancer patients.

CPT-11 (I; 7-ethyl-10-[4-(1-piperidino)-1- piperidino]carbonyloxycamptothecin) is a new anticancer agent currently under clinical development. A sensitive high-performance liquid chromatographic assay suitable for the simultaneous determination of I and its active metabolite SN-38 (II) in human plasma, and their preliminary clinical pharmacokinetics, are described. Plasma samples were processed using a solid-phase (C18) extraction step allowing mean recoveries of I, II and the internal standard camptothecin (III) of 84, 99 and 72%, respectively. The extracts were chromatographed on a C18 reversed-phase column with a mobile phase composed of acetonitrile, phosphate buffer and heptanesulphonic acid, with fluorescence detection. The calibration graphs were linear over a wide range of concentrations (1 ng/ml-10 micrograms/ml), and the lower limit of determination was 1 ng/ml for both I and II. The method showed good precision: the within-day relative standard deviation (R.S.D.) (5-1000 ng/ml) was 13.0% (range 4.9-19.4%) for I and 12.8% (6.7-19.1%) for II; the between-day R.S.D. (5-10,000 ng/ml was 7.9% (5.4-17.5%) for I and 9.7% (3.5-15.1%) for II. Using this assay, plasma pharmacokinetics of both I and II were simultaneously determined in three patients receiving 100 mg/m2 I as a 30-min intravenous infusion. The mean peak plasma concentration of I at the end of the intravenous infusion was 2400 +/- 285 ng/ml (mean +/- standard error of the mean). Plasma decay was triphasic with half-lives alpha, beta and gamma of 5.4 +/- 1.8 min, 2.5 +/- 0.5 h and 20.2 +/- 4.6 h, respectively. The volume of distribution at steady state was 105 +/- 15 l/m2, and the total body clearance was 12.5 +/- 1.9 l/h.m2. The maximum concentrations of the active metabolite II reached 36 +/- 11 ng/ml.     

Determination of thyreostatics in animal feed by micellar electrokinetic chromatography

The determination of the thyreostatics 2-thiouracil, its derivatives (4-methyl-2-thiouracil, 4-propyl-2-thiouracil and 4-phenyl-2-thiouracil) and methimazole in manufactured dried animal feed by micellar electrokinetic chromatography (MEKC) is described. A 99 +/- 5% extraction yield at the 20 micrograms g-1 level (n = 8) was achieved by shaking the milled fodder with methanol-1 M NaOH (80 + 20). Aliquots of the supernatant were injected in a 75 microns x 33.5 cm uncoated silica capillary using pressure; separation was performed at 23 degrees C with 15 kV (positive polarity) in a background electrolyte (BGE) containing 40 mM sodium dihydrogenphosphate, 50 mM sodium dodecyl sulfate and 15 mM Tween 20 at pH 9. When the surfactants were added to the BGE, all the thyreostatics were well resolved and the fodder extracts showed lower backgrounds. The peaks appeared within the 2.25-5.2 min range with efficiencies in the 2.5 x 10(4)-8 x 10(4) range; methimazole appeared in the vicinity of the electroosmotic migration time. Calibration curves were linear within the studied range (20-200 micrograms ml-1, r2 > 0.998). Limits of detection in the extracts of spiked fodder samples ranged from 0.25 to 0.4 microgram ml-1, which corresponded to 0.6-1.0 microgram of drug per gram of fodder. Peak area repeatabilities were about 4% at the 20 micrograms ml-1 level.     

Phytochemical investigation and hepatoprotective activity of Cupressus sempervirens L. leaves growing in Egypt

Three phenolic compounds cosmosiin, caffeic acid, and p-coumaric acid were isolated for the first time from the leaves of Cupressus sempervirens L., together with cupressuflavone, amentoflavone, rutin, quercitrin, quercetin, myricitrin. The isolated compounds were identified using (1)H- and (13)C-NMR spectra. The hepatoprotective activity of the MeOH extract was carried out in liver homogenate of normal and CCl(4)-treated rats; a significant decrease in glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, cholesterol level, and triglycerides, while a significant increase in the total protein level, was observed after the oral administration of MeOH extract. The free radical scavenging activity against stable 2,2-diphenyl-2-picrylhydrazyl (DPPH*) was measured for MeOH extract and some of the isolated phenolic compounds in comparison with alpha-tocopherol and butylated hydroxy toluene as standard antioxidants using ESR technique, showed high antioxidant activity for quercetin, rutin, caffeic acid, and p-coumaric acid.     

The hypotensive effect and antifungal activity of 3,3'-dihydroxy-alpha,beta-diethyldiphenylethane

3,3'-Dihydroxy-alpha,beta-diethyldiphenylethane (I), an isomer of hexestrol (II), was found to have a strong hypotensive effect on rats and antifungal activity against some pathogenic fungi. The hypotensive effect of I (-100.0 +/- 21.0 mmHg, 10 mg/kg, i.v.) was stronger than that of II (-40.0 +/- 2.60 mmHg, i.v.). Unlike II and other oxystilbene-related compounds already tested, I showed prolonged hypotensive action at the dose of 10 mg/kg, and the blood pressure was not restored to the original level within 20 min. I also showed antifungal activity against all fungi tested (minimal inhibitory concentration: 10-120 micrograms/ml). It should be emphasized that by shifting the phenolic hydroxyl group to m-position, the biological activities were greatly increased.     

In vitro antioxidant activities of five cultivars of daylily flowers from China

Different cultivars of functional vegetables may vary in an antioxidant capacity. In this study, the antioxidant activity of methanol extracts from five cultivars of daylily flower grown in China was compared by various antioxidant assays, including total antioxidant activity, free radical scavenging, superoxide anion radical scavenging and reducing power, the various antioxidant activities were compared to standard antioxidants such as butylated hydroxyanisole and ascorbic acid. All the extracts showed strong antioxidant activity in all tested methods. Among the five cultivars, Panlonghua and Mengzihua were found to possess the highest antioxidant activity, while Xiye and Baihua were found to possess the lowest antioxidant activity. In addition, a high positive correlation between the antioxidant properties and total phenolic content was observed.     

Determination of nickel in biological samples spectrophotometrically involving complexation reaction with some thiazolylazo compounds

Three new heterocyclic azo compounds, 5(2-benzothiazolylazo)2,2 biphenyldiol (I), 5-(2-benzothiazolylazo)-8-hydroxyquinolene (II) and 4-(2-benzothiazo-lylazo)3-hydroxy-2-naphthoic acid (III) were synthesized. The formation constant of the reagents in 30% (v/v) ethanol and reactions with various metal ions were studied. These reagents (I-III) reacts with nickel ion to form (2:1) colored complex with an absorption band at 604, 635 and 643 nm. The apparent stability constants and the optimum conditions for complete color development were investigated. Beer's law is obeyed over the concentration ranges of 0.05-3.50, 0.05-4.00 and 0.05-3.10 micrograms ml-1. For more accurate analyses, Ringbom optimum concentration ranges were found to be 0.20-3.25, 0.20-3.80 and 0.20-2.90 micrograms ml-1 using reagents I, II and III, respectively. The apparent molar absorptivity and Sandell sensitivity were also calculated. The interference of various foreign ions on the determination of nickel was investigated. The proposed method has been successfully applied to the determination of nickel substrates in various biological samples.     

Identification of major phenolic compounds of Chinese water chestnut and their antioxidant activity

Chinese water chestnut (CWC) is one of the most popular foods among Asian people due to its special taste and medical function. Experiments were conducted to test the antioxidant activity and then determine the major phenolic compound components present in CWC. CWC phenolic extract strongly inhibited linoleic acid oxidation and exhibited a dose-dependent free-radical scavenging activity against alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radicals, superoxide anions and hydroxyl radicals, which was superior to ascorbic acid and butylated hydroxytoluene (BHT), two commercial used antioxidants. Furthermore, the CWC extract was found to have a relatively higher reducing power, compared with BHT. The major phenolic compounds present in CWC tissues were extracted, purified and identified by high-performance liquid chromatograph (HPLC) as (-)-gallocatechin gallate, (-)-epicatechin gallate and (+)-catechin gallate. This study suggests that CWC tissues exhibit great potential for antioxidant activity and may be useful for their nutritional and medicinal functions.     

Determination of beta-lactams and their biosynthetic intermediates in fermentation media by pre-column derivatisation followed by fluorescence detection

This paper describes a novel and sensitive pre-column derivatisation method for the detection and quantitation of beta-lactams and their biosynthetic precursors at trace levels in fermentation media. Filtered broths from fermentations of strains of Penicillium chrysogenum and Cephalosporium acremonium, after deproteination and centrifugation, were incubated with 9-fluorenylmethylchloroformate for 5 min at 20 degrees C in 0.2 M borate buffer at pH 7.7. Following two-fold pentane extraction of the reagent hydrolysis product, the aqueous layer was injected directly onto a C18 reversed-phase column, and products were detected spectrofluorimetrically with excitation and emission wavelengths of 260 and 313 nm, respectively. Detection limits of 0.01 and 0.05 micrograms ml-1 were achieved for both 6-aminopenicillanic acid (6-APA) and isopenicillin N in borate buffer and filtered fermentation broths, respectively, using a 10-microliter injection volume. A linear calibration for 6-APA in fermentation broth was obtained for a very wide concentration range (0.05-100 micrograms ml-1). Detection limits for solutions of cephalosporin C, deacetylcephalosporin C and deacetoxycephalosporin C in broth were all 0.25 micrograms ml-1. The detection limit for the beta-lactam precursor delta-(L-aminoadipyl)-L-alpha-cysteinyl-D-valine (ACV) dimer in borate buffer was 0.5 microgram ml-1. The cephalosporins and ACV dimer gave linear plots in the ranges 3-25 and 1-100 micrograms ml-1, respectively. Repeated analysis of 6-APA at a concentration of 10 micrograms ml-1 in filtered broth gave a mean peak area of 2.5.10(6) with a standard deviation of 2.6.10(5) using a 10-microliter injection volume. Ampicillin spiked into deproteinated blood serum gave a linear calibration in the concentration range 2-100 micrograms ml-1.     

Spectrophotometric determination of certain cephalosporins using molybdophosphoric acid. Part II. Determination of cefadroxil, cefapirin, ceforanide and cefuroxime

The use of molybdophosphoric acid as an oxidising agent for the spectrophotometric determination of four cephalosporin derivatives, viz., cefadroxil monohydrate (I), cefapirin sodium (II), ceforanide L-lysine (III) and cefuroxime sodium (IV), either in the pure form or in pharmaceutical formulations is described. Beer's law is obeyed up to 100 micrograms ml-1 for I, up to 60 micrograms ml-1 for II and IV and up to 80 micrograms ml-1 for III. The molar absorptivities were 4.58 X 10(3), 11.3 X 10(3), 9.8 X 10(3) and 10.9 X 10(3) l mol-1 cm-1 and the Sandell sensitivities were 83.3, 39.3, 53.0 and 41.0 ng cm-2 for I, II, III and IV, respectively. The slopes and intercepts of the equations of the regression line were calculated for each of these drugs with the following correlation coefficients: I, 0.9993; II, 0.9999; III, 1.000; and IV, 0.9999. These antibiotics were determined successfully both in the pure form and in pharmaceutical preparations. The results demonstrated that the proposed procedure is at least as accurate, precise and reproducible as the official methods, while being simpler and less time consuming. A statistical analysis indicated that there was no significant difference between the results obtained by the proposed procedure and those of the official methods.     

Purification and Identification of Novel Antioxidant Peptides from Enzymatically Hydrolysed Samia ricini Pupae

The emergence of excessive free radicals leads to the destruction of various systems within the body. These free radicals also affect nutritional values, color, taste, and emit an odor akin to rancid food. Most food industries use synthetic antioxidants, such as BHT (butylated hydroxytoluene) or BHA (butylated hydroxy anisole). However, high doses of these can be harmful to our health. Therefore, an antioxidant compounds, such as bioactive peptides from edible animals or plants, have emerged to be a very promising alternative as they reduce potential side effects. This study focused on the purification and identification of antioxidant peptides from protein hydrolysates of wild silkworm pupae (Samia ricini). Antioxidant peptides were purified from the hydrolysate by ultrafiltration and RP-HPLC. The results showed that protein hydrolysate from S. ricini pupae by trypsin with a molecular weight lower than 3 kDa and highly hydrophobic property, exhibited strong DPPH radical scavenging activity and chelating activity. Further identification of peptides from the fraction with the highest antioxidant activity was carried out using LC-MS/MS. Three novel peptides, i.e., Met-Ley-Ile-Ile-Ile-Met-Arg, Leu-Asn-Lys-Asp-Leu-Met-Arg, and Glu-Asn-Ile-Ile-Leu-Phe-Arg, were identified. The results of this study indicated that the protein hydrolysate from S. ricini pupae possessed potent biological activity, and the novel antioxidant peptides could be utilized to develop health-related antioxidants in food industry.     

A highly sensitive spectrophotometric method for the determination of some phenothiazine antipsychotics using chloramine-T and indigocarmine.

A new simple, accurate and precise spectrophotometric method for the determination of six phenothiazine drugs in pure form and in dosage forms is described. The method is based on the oxidation of the studied drugs by a known excess of Chloramine-T in hydrochloric acid medium and subsequent determination of the unreacted oxidant by reacting it with indigocarmine in the same acid medium. The reacted oxidant corresponds to the drug content. The colored species exhibits maximum absorption at 610 nm. The apparent molar absorptivity values and Sandell sensitivity values are in the range 1.53 x 10(4)-2.96 x 10(4) l mol-1 cm-1 and 13.75-37.15 ng cm-2, respectively. The method is highly sensitive and suitable for 1-15 micrograms ml-1 concentrations with the detection limits being in the range, 0.0651-0.1724 microgram ml-1. The method was successfully applied to the studied drugs in their dosage forms. The results are reproducible within +/- 1% and compare favorably with those obtained by the procedures of the British Pharmacopeia.     

Evaluation of antioxidant activity of isoferulic acid in vitro

Isoferulic acid (3-hydroxy-4-methoxycinnamic acid, IFA), the isomer of ferulic acid (4-hydroxy-3-methoxycinnamic acid), is a rare phenolic acid occurring in Rhizoma Cimicifugae. Unlike ferulic acid, which has been well investigated, the antioxidant activity of IFA has not been measured. In this study, IFA was systematically evaluated for its in vitro antioxidant activity for the first time. IC50 values were calculated of 7.30 +/- 0.57, 4.58 +/- 0.17, 1.08 +/- 0.01, 8.84 +/- 0.43, 7.69 +/- 0.39, 1.57 +/- 0.2, 13.33 +/- 0.49 microg/mL, respectively, for lipid peroxidation, DPPH (1,1-diphenyl-2-picrylhydrazyl radical) and ABTS (3-ethylbenzthiazoline-6-sulfonic acid diammonium salt) radical scavenging, reducing power on Fe3+ and CU2+ ions, and hydroxyl and superoxide anion radical scavenging. Comparison with the IC50 values with those of the positive controls, Trolox and butylated hydroxyanisole (BHA), it can be concluded that isoferulic acid is an effective natural antioxidant in both lipid and aqueous media.     

Determination of Synthetic Phenolic Antioxidants in Vegetable Oil and Oil-Enriched Foods by High-Performance Liquid Chromatography with Electrochemical Detection

Three synthetic phenolic antioxidants, tertiary butyl hydroquinone, butylated hydroxytoluene, and butylated hydroxyanisole, were determined in vegetable oil and oil-enriched food by high-performance liquid chromatography (HPLC) with electrochemical detection. The separation was achieved using a reverse-phase column and gradient elution with methanol and 1% acetic acid. The limits of detection and quantification of the analytes were 2–120 lower, higher than those obtained by diode-array detection. The recoveries were 103.3% for tertiary butyl hydroquinone, 97.3% for butylated hydroxyanisole, and 95.2% for butylated hydroxytoluene. The results showed that HPLC with electrochemical detection is suitable for the quantification of low concentrations of phenolic antioxidants in vegetable oil and oil-enriched food with high sensitivity and accuracy.     

Photoprotective Potential of Baccharis antioquensis (Asteraceae) as Natural Sunscreen

In the quest for new natural agents of photoprotection, we evaluated the photoprotective and antioxidant activity of B. antioquensis leaf extracts as well as its phenolic composition. The methanolic extract treated with activated carbon showed the highest absorption coefficients for UVA‐UVB radiation, as well as an antioxidant capacity comparable to butylated hydroxy toluene. Furthermore, the formulation containing this extract showed suitable sensorial and photostable characteristics for topical use, and significant values of UVAPF, critical wavelength (λ_c), UVA/UVB ratio and sun protection factor (5.3, 378 nm, 0.78 and 9.1 ± 0.1, respectively). In addition, three glycoside derivatives of quercetin, a kaempferol glycoside and a derivative of caffeic acid were the main polyphenolic compounds identified. These results demonstrate the potential of B. antioquensis extracts to be used as active components of novel, natural sunscreens.     

Determination of antioxidants by a novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) assay with post-column detection

A novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) method was developed for the selective determination of polyphenols (flavonoids, simple phenolic and hydroxycinnamic acids) in complex plant matrices. The method combines chromatographic separation, constituent analysis, and post-column identification of antioxidants in plant extracts. The separation of polyphenols was performed on a C18 column using gradient elution with two different mobile phase solutions, i.e., MeOH and 0.2% o-phosphoric acid. The HPLC-separated antioxidant polyphenols in the extracts react with copper(II)-neocuproine (Cu(II)-Nc) reagent in a post-column reaction coil to form a derivative. The reagent is reduced by antioxidants to the copper(I)-neocuproine (Cu(I)-Nc) chelate having maximum absorption at 450 nm. The negative peaks of antioxidant constituents were monitored by measuring the increase in absorbance due to Cu(I)-Nc. The detection limits of polyphenols at 450 nm (in the range of 0.17-3.46 μM) after post-column derivatization were comparable to those at 280 nm UV detection without derivatization. The developed method was successfully applied to the identification of antioxidant compounds in crude extracts of Camellia sinensis, Origanum marjorana and Mentha. The method is rapid, inexpensive, versatile, non-laborious, uses stable reagents, and enables the on-line qualitative and quantitative estimation of antioxidant constituents of complex plant samples.     

Electrochemical oxidation at carbon paste electrode of tacrine and 1-hydroxytacrine and differential pulse voltammetric determination of tacrine in pharmaceuticals and human urine

The electrochemical oxidation of tacrine and its 1-OH-metabolite, has been studied by cyclic voltammetry and differential pulse voltammetry by using carbon paste electrodes. The peak current-concentration relationship was found to be linear up to 20 micrograms ml-1 with detection limits of 0.06 microgram ml-1 for tacrine and 0.18 microgram ml-1 for 1-OH-tacrine and quantitation limits of 0.20 microgram ml-1 for tacrine and 0.37 microgram ml-1 for 1-OH-tacrine. A method for determining tacrine by differential pulse voltammetry in pharmaceuticals and human urine, in the presence of 1-OH-tacrine, has been developed.     

Simultaneous determination of benzoic acid and saccharin in soft drinks by using lanthanide-sensitized luminescence

A simple and fast approach is used for the first time to develop a time resolved lanthanide-sensitized luminescence method for the simultaneous determination of a preservative and a sweetener, namely benzoic acid (BZ) and saccharin (SC), respectively, in food samples. The method involves the formation of the corresponding ternary chelates with terbium(III) and trioctylphosphine oxide (TOPO) in the presence of Triton X-100, and the measurement of the initial rate and equilibrium signal of this system, which were obtained in 0.1 and 5 s, respectively. The dynamic ranges of the calibration graphs, obtained by using kinetic and equilibrium measurements, were 0.2-36 micrograms ml-1 and 0.15-30 micrograms ml-1, respectively, for BZ, and 3.3-24 micrograms ml-1 and 4-36 micrograms ml-1 for SC and the detection limits were 0.07 and 0.04 microgram ml-1, respectively, for BZ, and 1.1 and 1.2 micrograms ml-1, respectively, for sodium SC. The relative standard deviation ranged between 2.3 and 3.0%. Both compounds were determined simultaneously by using a system of two equations which were resolved by using the calibration data obtained individually for each analyte and by multiple linear regression. Mixtures of BZ and SC in ratios between 3:1 and 1:9 were satisfactorily resolved by using both approaches. The method was applied to the direct analysis of several soft drinks. Analytical recoveries ranged between 89.3 and 108.5%.     

Synthesis,Characterization, and Antioxidant Evaluations of New 2‐Oxochromene and Benzofuran Derivatives Catalyzed by KF/CP

An eco‐friendly one‐pot procedure has been developed for the preparation of novel 2‐oxochromene and benzofuran derivatives through the sequential addition/intramolecular cyclization reactions of 1‐(6‐hydroxy‐2‐isopropenyl‐1‐benzofuran‐5‐yl)‐1‐ethanone, activated acetylenic compounds, and triphenylphosphine in the presence of catalytic amount of potassium fluoride impregnated clinoptilolite nanocatalyst in water at room temperature. The antioxidant activities of the newly synthesized derivatives ( 4a , 4b , and 5a ) were also screened by diphenyl‐2‐picrylhydrazyl radical scavenging and ferric ion reducing potential assays compared with the synthetic antioxidants (2‐tert‐butylhydroquinone and butylated hydroxytoluene). Compounds 4a and 4b were shown good antioxidant activities.     

Kinetic determination of tellurium based on its inhibitory effect on the palladium(II)-catalysed reaction between pyronine G and hypophosphite ion

A kinetic method for the determination of Te based on its inhibitory effect on the PdII-catalysed reaction between pyronine G and H2PO2- is described. The influence of experimental variables on the rate of the process and the potential interfering effect of a large number of ions has been studied. Under the selected experimental conditions: 6 x 10(-5) M pyronine G; 0.6 M H2PO2-; pH 2.6, adjusted with Britton-Robinson buffer; 0.80 microgram ml-1 of PdII; and a temperature of 22 +/- 0.2 degrees C, Te was determined in the concentration range 0.08-0.85 microgram ml-1. The method was applied to the determination of Te in waters and lead concentrates.