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Diode Array Detection (diode + array_detection)
Selected AbstractsEvaluation of lignans and free and linked hydroxy-tyrosol and tyrosol in extra virgin olive oil after hydrolysis processesJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 5 2006Nadia Mulinacci Abstract We describe chemical hydrolytic procedures to evaluate the total amount of tyrosol and hydroxy-tyrosol free and/or linked to secoiridoidic molecules (acid hydrolysis). At the same time a rapid determination of the lignans in complex minor polar compound (MPC) extracts is proposed (alkaline hydrolysis). High-performance liquid chromatography/diode array detection (HPLC/DAD) and HPLC/MS were applied as reference methods to evaluate the quantitative results from the hydrolysis experiments. The optimized acid hydrolysis procedures were first applied to an oleuropein standard and then to MPC fractions extracted from several commercial extra virgin olive oils. The results confirm the applicability of the method, consisting in the acid hydrolysis of complex mixtures of secoiridoidic derivatives, to determine the antioxidant potential in terms of MPC. These data can contribute to forecasting the potential ageing resistance of an extra virgin olive oil in terms of antioxidant potency. Finally, alkaline hydrolysis allows confirmation and easy determination of the amount of lignans, especially in those MPC fractions which are particularly complex. Copyright © 2006 Society of Chemical Industry [source] Comparative study of six pear cultivars in terms of their phenolic and vitamin C contents and antioxidant capacityJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2003Andrea C Galvis Sánchez Abstract The main phenolic compounds in six pear cultivars were identified and quantified using high-performance liquid chromatography/diode array detection (HPLC/DAD) and HPLC/electrospray ionisation mass spectrometry (HPLC/ESIMS). Major quantitative differences were found in the phenolic profiles. The peel contained higher concentrations of chlorogenic acid, flavonols and arbutin than the flesh, where only chlorogenic acid was detected. Total phenolics ranged from 1235 to 2005 mg kg,1 in the peel and from 28 to 81 mg k g,1 in the flesh. Ascorbic acid and dehydroascorbic acid were detected in the peel, whereas only dehydroascorbic acid was present in the flesh. The ranges of vitamin C content were from 116 to 228 mg kg,1 in the peel and from 28 to 53 mg kg,1 in the flesh. The antioxidant capacity was correlated with the content of chlorogenic acid (r = 0.46), while ascorbic acid made only a small contribution to the total antioxidant capacity of the fruit. Copyright © 2003 Society of Chemical Industry [source] In vivo Skin Irritation Potential of a Castanea sativa (Chestnut) Leaf Extract, a Putative Natural Antioxidant for Topical ApplicationBASIC AND CLINICAL PHARMACOLOGY & TOXICOLOGY, Issue 5 2008Isabel F. Almeida However, natural products can provoke skin adverse effects, such as allergic and irritant contact dermatitis. Skin irritation potential of Castanea sativa leaf ethanol:water (7:3) extract was investigated by performing an in vivo patch test in 20 volunteers. Before performing the irritation test, the selection of the solvent and extraction method was guided by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) free radical scavenging test and polyphenols extraction (measured by the Folin Ciocalteu assay). Iron-chelating activity and the phenolic composition (high performance liquid chromatography/diode array detection) were evaluated for the extract obtained under optimized conditions. The extraction method adopted consisted in 5 short extractions (10 min.) with ethanol:water (7:3), performed at 40°. The IC50 found for the iron chelation and DPPH scavenging assays were 132.94 ± 9.72 and 12.58 ± 0.54 µg/ml (mean ± S.E.M.), respectively. The total phenolic content was found to be 283.8 ± 8.74 mg GAE/g extract (mean ± S.E.M.). Five phenolic compounds were identified in the extract, namely, chlorogenic acid, ellagic acid, rutin, isoquercitrin and hyperoside. The patch test carried out showed that, with respect to irritant effects, this extract can be regarded as safe for topical application. [source] Data processing in metabolic fingerprinting by CE-UV: Application to urine samples from autistic childrenELECTROPHORESIS, Issue 6 2007Ana C. Soria Abstract Metabolic fingerprinting of biofluids such as urine can be used to detect and analyse differences between individuals. However, before pattern recognition methods can be utilised for classification, preprocessing techniques for the denoising, baseline removal, normalisation and alignment of electropherograms must be applied. Here a MEKC method using diode array detection has been used for high-resolution separation of both charged and neutral metabolites. Novel and generic algorithms have been developed for use prior to multivariate data analysis. Alignment is achieved by combining the use of reference peaks with a method that uses information from multiple wavelengths to align electropherograms to a reference signal. This metabolic fingerprinting approach by MEKC has been applied for the first time to urine samples from autistic and control children in a nontargeted and unbiased search for markers for autism. Although no biomarkers for autism could be determined using MEKC data here, the general approach presented could also be applied to the processing of other data collected by CE with UV,Vis detection. [source] Metabolism of fluoranthene in different plant cell cultures and intact plantsENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 5 2000Marit Kolb Abstract The metabolism of fluoranthene was investigated in 11 cell cultures of different plant species using a [14C]-labeled standard. Most species metabolized less than 5% of fluoranthene to soluble metabolites and formed less than 5% nonextractable residues during the standardized 48-h test procedure. Higher metabolic rates were observed in lettuce (Lactuca sativa, 6%), wheat (Tricitum aestivum, 9%), and tomato (Lycopersicon esculentum, 15%). A special high metabolic rate of nearly 50% was determined for the rose species Paul's Scarlet. Chromatographic analysis of metabolites extracted from aseptically grown tomato plants proved that the metabolites detected in the cell cultures were also formed in the intact plants. Metabolites produced in tomato and rose cells from [14C]-fluoranthene were conjugated with glucose, glucuronic acid, and other cell components. After acid hydrolyses, the main metabolite of both species was 1-hydroxyfluoranthene as identified by gas chromatography,mass spectrometry and high-performance liquid chromatography with diode array detection. The second metabolite formed by both species was 8-hydroxy-fluoranthene. A third metabolite in tomatoes was 3-hydroxyfluoranthene. [source] Hard-modelled trilinear decomposition (HTD) for an enhanced kinetic multicomponent analysisJOURNAL OF CHEMOMETRICS, Issue 5 2002Yorck-Michael Neuhold Abstract We present a novel approach for kinetic, spectral and chromatographic resolution of trilinear data sets acquired from slow chemical reaction processes via repeated chromatographic analysis with diode array detection. The method is based on fitting rate constants of distinct chemical model reactions (hard-modelled, integrated rate laws) by a Newton,Gauss,Levenberg/Marquardt (NGL/M) optimization in combination with principal component analysis (PCA) and/or evolving factor analysis (EFA), both known as powerful methods from bilinear data analysis. We call our method hard-modelled trilinear decomposition (HTD). Compared with classical bilinear hard-modelled kinetic data analysis, the additional chromatographic resolution leads to two major advantages: (1) the differentiation of indistinguishable rate laws, as they can occur in consecutive first-order reactions; and (2) the circumvention of many problems due to rank deficiencies in the kinetic concentration profiles. In this paper we present the theoretical background of the algorithm and discuss selected chemical rate laws. Copyright © 2002 John Wiley & Sons, Ltd. [source] EFFECTS OF DIFFERENT MACERATION TIMES AND PECTOLYTIC ENZYME ADDITION ON THE ANTHOCYANIN COMPOSITION OF VITIS VINIFERA CV. KALECIK KARASI WINESJOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 3 2009HASIM KELEBEK ABSTRACT Kalecik karasi is an important red grape cultivar for winemaking in Turkey. The effect of three different maceration times (3, 6 and 12 days) and addition of pectolytic enzyme (2 and 4 g/hL) on the anthocyanin and chemical composition of Kalecik karasi wines were studied. High performance liquid chromatography-mass spectrometry coupled with diode array detection was used for analysis. Fourteen anthocyanin compounds were detected in wines. Major anthocyanins in all wines are malvidin-3-glucoside and its acylated esters. The results showed that increasing maceration time, especially with addition of enzymes, gives significant increases in anthocyanin contents. Moreover, the wines treated with enzymes had higher values in total phenolics, tannins, and color intensity than the control wines. PRACTICAL APPLICATIONS Anthocyanins are the most important polyphenols in red grapes and red wines with potential health benefits. Therefore, the first analysis of the anthocyanins contents of wine obtained from important turkish cv. Kalecik karasi using liquid-chromatography-mass spectrometry and the influence of different maceration times and addition of pectolytic enzyme on these important phenolic compounds are of interest for scientific literature, the wine industry as well as for the wine consumer. [source] Direct characterization of aqueous extract of Hibiscus sabdariffa using HPLC with diode array detection coupled to ESI and ion trap MSJOURNAL OF SEPARATION SCIENCE, JSS, Issue 20 2009Inmaculada C. Rodríguez-Medina Abstract The phenolic fraction and other polar compounds of the Hibiscus sabdariffa were separated and identified by HPLC with diode array detection coupled to electrospray TOF and IT tandem MS (DAD-HPLC-ESI-TOF-MS and IT-MS). The H. sabdariffa aqueous extract was filtered and directly injected into the LC system. The analysis of the compounds was carried out by RP HPLC coupled to DAD and TOF-MS in order to obtain molecular formula and exact mass. Posterior analyses with IT-MS were performed and the fragmentation pattern and confirmation of the structures were achieved. The H. sabdariffa samples were successfully analyzed in positive and negative ionization modes with two optimized linear gradients. In positive mode, the two most representative anthocyanins and other compounds were identified whereas the phenolic fraction, hydroxycitric acid and its lactone were identified using the negative ionization mode. [source] Determination of uric acid in plasma and allantoic fluid of chicken embryos by capillary electrophoresisJOURNAL OF SEPARATION SCIENCE, JSS, Issue 12 2007Jana Mat, ková Abstract Capillary electrophoresis with diode array detection (DAD) was used to determine uric acid (UA) in chicken plasma and the allantoic fluid of chicken embryos. Complete separation of uric and ascorbic acids was attained in less than 10 min in the optimized BGE containing 60 mM MES + 30 mM Tris + 0.001% (w/v) polybrene (pH 6.1). The limit of UA detection (0.2 mg/L) was found to be low enough for sensitive analysis of native plasma and allantoic fluid samples. Range of linearity (1,200 mg/L), repeatability for peak area (CV <4.1%) and migration time (CV <2.5%), as well as recovery of UA from biological samples (97,100%), were found to be satisfactory. The method was applied to detect the elevated UA concentrations (hyperuricemia) in chicken embryos with induced unilateral renal agenesis. CE/DAD analysis of the chicken plasma can be carried out with a relatively small volume of samples (1 ,L). [source] Compositional changes induced by UV-B radiation treatment of common bean and soybean seedlings monitored by capillary electrophoresis with diode array detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 4 2007Giovanni Dinelli Abstract In this work, a new CE method with diode array detection (DAD) was developed for the monitoring and quantitation of flavonoids in different beans treated and untreated with UV-B radiation. Flavonoid concentration was monitored in UV-B-treated and untreated sprouts of three common beans (Zolfino ecotype, cv. Verdone, cv. Lingua di Fuoco) and one soybean (cv. Pacific). After acid hydrolysis of extracts, the CE-DAD method provides reproducible quantitative determinations of daidzein, glycitein, genistein, and kaempferol at ppm level in these natural matrices within a relatively short time (less than 16 min). Total flavonoid content determined by CE-DAD was 159 ± 8, 26 ± 2, 13 ± 1, and 1.3 ± 0.3 ,g/g fresh weight for untreated sprouts of Pacific soybean, Verdone bean, Zolfino bean, and Lingua di Fuoco bean, respectively. UV-B treatment caused no significant quantitative effect on Pacific soybean sprouts, whereas it enhanced the total isoflavone content by 1.5, 1.8, and 3.2-fold in Verdone, Zolfino, and Lingua di Fuoco beans, respectively. The proposed method shows (i) the potentialities of bean sprouts as a natural source of bioactive compounds (antioxidants); (ii) the technological role of UV-B treatment for sprout isoflavone enrichment; and (iii) the good capabilities of CE-DAD to monitor this process. [source] Simultaneous analysis of nine active components in Gegen Qinlian preparations by high-performance liquid chromatography with diode array detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2006Lihong Chen Abstract HPLC with diode array detection (HPLC/DAD) was employed to determine the quantities of puerarin, daidzin, daidzein, berberine, palmatine, coptisine, baicalin, baicalein, and glycyrrhizin in Gegen Qinlian preparations of three different pharmaceutical forms including decoction, dispensing granule and pill. The calibration curves for the nine bioactive components were linear in the given concentration ranges. The precision of the method was in the range of 0.2,5.0% (RSD), and the recoveries of this method were between 96.5 and 104.1%. The proposed method was applicable to analyze Gegen Qinlian preparations. [source] Simultaneous determination of carotenoids, tocopherols, and ,-oryzanol in crude rice bran oil by liquid chromatography coupled to diode array and mass spectrometric detection employing silica C30 stationary phasesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2005Wolfgang Stöggl Abstract Crude rice bran oil contains tocopherols (vitamin E), carotenoids (vitamin A), and phytosterols, which possess antioxidant activities and show promising effects as preventive and therapeutic agents. The aim of this work was to establish methods and to compare C18 and C30 silica stationary phases in order to separate and detect tocopherols, carotenoids, and ,-oryzanol in one single run. Comparing RP-LC on silica C18 and C30, higher resolution between all target compounds was obtained using the C30 stationary phase. Methanol was used as eluent and the elution strength was increased by the addition of tert -butyl methyl ether for highly hydrophobic analytes such as ,-oryzanol. Detection was accomplished by diode array detection from 200 to 500 nm. Absorbance maxima were found at 295 nm for tocopherols, 324 nm for ,-oryzanol, and 450 nm for carotenoids. Furthermore, compounds were characterized and identified on the basis of their UV-spectra. Both RP systems were coupled to MS (LC-MS) by using an atmospheric pressure chemical ionization interface. [source] Characterization via liquid chromatography coupled to diode array detector and tandem mass spectrometry of supercritical fluid antioxidant extracts of Spirulina platensis microalgaJOURNAL OF SEPARATION SCIENCE, JSS, Issue 9-10 2005Jose A. Mendiola Abstract Spirulina platensis microalga has been extracted on a pilot scale plant using supercritical fluid extraction (SFE) under various extraction conditions. The extraction yield and the antioxidant activity of the extracts were evaluated in order to select those extracts with both the highest antioxidant capacity and a good extraction yield. These extracts were characterized using LC coupled to diode array detection (DAD) and LC coupled to mass spectrometry (MS) with two different interfaces, atmospheric pressure chemical ionization (APCI) and electrospray (ESI) which allowed us to perform tandem MS by using an ion trap analyzer. The best extraction conditions were as follows: CO2 with 10% of modifier (ethanol) as extraction solvent, 55°C (extraction temperature) and 220 bar (extraction pressure). Fractionation was achieved by cascade depressurization providing two extracts with different activity and chemical composition. Several compounds have been identified in the extracts, corresponding to different carotenoids previously identified in Spirulina platensis microalga along with chlorophyll a and some degradation products. Also, the structure of some phenolic compounds could be tentatively identified. The antioxidant activity of the extracts could be attributed to some of the above mentioned compounds. [source] A comparative study of several HPLC methods for determining free amino acid profiles in honeyJOURNAL OF SEPARATION SCIENCE, JSS, Issue 9-10 2005José Luis Bernal Abstract A study of the viability of three derivatizing reagents for obtaining amino acid profiles in honey through high performance liquid chromatography (HPLC) is presented. A method using diode array detection based on a reaction with diethyl ethoxymethylene malonate (DEMM) and two other methods using fluorescence detection based on derivatization with fluorenylmethyl chloroformate (FMOC-Cl) and 6-aminoquinolyl- N -hydroxysuccinimidyl carbamate (AQC) have been developed. The three methods yield detection limits close to the ppb level, but vary in relation to other analytical characteristics. The use of methyl chloroformate derivatives allows the profile to be obtained with the greatest sensitivity within a short time frame. On applying such methods to honey samples of diverse botanical origin, we observe that the proline values obtained are always lower than those found using the official spectrophotometric method, thereby underlining the advisability of using HPLC methods to reduce uncertainty in these results. [source] Determination of cyprodinil and fludioxonil in the fermentative process of must by high-performance liquid chromatography,diode array detectionJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 11 2008Luis Vaquero-Fernández Abstract BACKGROUND: A quantitative, selective and sensitive high-performance liquid chromatographic method is described for the analysis of new fungicides cyprodinil, fludioxonil and their commercial formulation Switch in model solutions of must and wine, as well as samples during alcoholic fermentation. A study of the dissipation of residues was carried out. RESULTS: The proposed method is based on liquid,liquid extraction (LLE) followed by high-performance liquid chromatography and diode array detection. Dichloromethane was the most appropriate solvent for extracting cyprodinil and fludioxonil in samples. Quality parameters of the proposed method presented good recovery (ca. 97% for almost all compounds) and precision (between 4.8% and 5.4%), and limits of quantification were lower than maximum residue limits (MRLs) in grapes. CONCLUSIONS: There is no matrix effect in the analysis of cyprodinil and fludioxonil. The application of the fermentative process on cyprodinil and fludioxonil fungicides causes a decrease in the concentrations of these compounds. This decrease is slightly higher, the higher the initial concentration, without observing the appearance of any product in degradation. Fludioxonil shows a higher reduction when the compounds are presented together in Switch. Copyright © 2008 Society of Chemical Industry [source] High-performance liquid chromatography with diode array detection coupled to electrospray time-of-flight and ion-trap tandem mass spectrometry to identify phenolic compounds from a Cistus ladanifer aqueous extractPHYTOCHEMICAL ANALYSIS, Issue 4 2010S. Fernández-Arroyo Abstract Introduction , Cistus ladanifer is an aromatic shrub that is widespread in the Mediterranean region. The labdanum exudate is used in the fragrance industry and has been characterised. However, there is not enough information about the phenolic content of the raw plant, the aerial part of it being a very rich source of bioactive compounds. Objective , Characterisation of the bioactive compounds of the raw plant and its aerial parts. Methodology , High-performance liquid chromatography with diode array and electrospray ionisation mass spectrometric detection was used to carry out the comprehensive characterisation of a Cistus ladanifer shrub aqueous extract. Two different MS techniques were coupled to HPLC: time-of-flight mass spectrometry and tandem mass spectrometry. Results , Many well-known compounds present in Cistus ladanifer were characterised, such as flavonoids, phenolic acids, ellagitanins, hexahydroxydiphenoyl and derivatives, and other compounds. Conclusion , The method described simultaneously separated a wide range of phenolic compounds and the proposed characterisation of the major compounds of this extract was carried out. It is important to highlight that, to our knowledge, this is the first time that a Cistus ladanifer aqueous extract from the raw plant has been characterised. Copyright © 2009 John Wiley & Sons, Ltd. [source] High-performance liquid chromatography with electrospray ionisation mass spectrometry and diode array detection in the identification and quantification of the degradation products of calix[4]arene crown-6 under radiolysisRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 13 2004C. Lamouroux The extraction of 135Cs from high-activity liquid waste, arising from reprocessing of spent nuclear fuel, can be achieved by using calix[4]arene crown-6 compounds. The radiolytic degradation of di(n-octyloxy)calix[4]arene crown-6 (octMC6), in aliphatic or aromatic solvent in contact with 3 M nitric acid, was studied by high-performance liquid chromatography directly coupled to electrospray ionisation mass spectrometry (LC/ESI-MS). More than 50 distinct degradation products were observed, and about 30 of these were identified. These compounds can be assigned to three categories, namely, products of reactions involving radical cleavage or addition, of oxidation reactions, or of aromatic substitution reactions. The major product, corresponding to substitution by an NO2 group, was quantified by external standard calibration using a purified synthetic sample. Despite the observation of all these degradation compounds, octMC6 appears to be remarkably stable under these drastic conditions, combining hydrolysis (HNO3 3,M) and an extreme exposure to radiolysis (106,Gy). Less than 35% degradation of octMC6 was observed in aromatic solvent under these conditions. Copyright © 2004 John Wiley & Sons, Ltd. [source] Fragmentation study of iridoid glycosides and phenylpropanoid glycosides in Radix Scrophulariae by rapid resolution liquid chromatography with diode-array detection and electrospray ionization time-of-flight mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 8 2010Qian Wu Abstract Rapid resolution liquid chromatography (RRLC) coupled with diode array detection (DAD) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF MS) method was applied to the mass spectral study of a series of naturally occurring iridoid glycosides and phenylpropanoid glycosides in Radix Scrophulariae, which provides higher speed and increased sensitivity without loss of resolution. With dynamic adjustment as the key role of the fragmentor voltage and confirmed with authentic standards, valuable structural information regarding the nature of both the glycoside skeletons was thus obtained. Most compositions were found to possess organic acid moiety such as cinnamoyl, caffeoyl and ferulyol. Besides extensive fragmentation of the carbohydrate moiety, losses of the hydroxyl and glucose residue units showed in the spectra, permitting the exploration of the skeleton and the identity of substituents in the molecule. Ten major iridoid glycosides and 10 phenylpropanoid glycosides were identified or tentatively characterized based on their retention times, UV and TOF MS data. The major fragmentation pathways of PGs in Radix Scrophulariae obtained through the MS data was schemed systematically for the first time, which provides a reference for other PGs derivatives. Copyright © 2009 John Wiley & Sons, Ltd. [source] Quality assessment of Cortex Phellodendri by high-performance liquid chromatography coupled with electrospray ionization mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 4 2010Yong Mei Hu Abstract A simple method based on liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometry (LC-DAD-ESI-MS) was developed for the quality assessment of Cortex Phellodendri (CP), which was mainly derived from two species of Phellodendron chinense Schneid and Phellodendron amurense Rupr. Total 41 compounds, including 14 phenols, 24 alkaloids and three liminoidal triterpenes were identified or tentatively characterized from the 75% methanol extract of CP samples by online ESI-MSn fragmentation and UV spectra analysis. Among them, two phenols and six alkaloids were simultaneously quantified using HPLC-DAD method. The validated HPLC-DAD method showed a good linearity, precision, repeatability and accuracy for the quantification of eight marker compounds. Furthermore, the plausible fragmentation pathway of the representative compounds were proposed in the present study. The differences of the chemical constituents content and the comprehensive HPLC profiles between the two CP species using LC-DAD-ESI-MS method are reported for the first time, indicating that the CP drugs from different resources should be used separately in the clinic. Copyright © 2009 John Wiley & Sons, Ltd. [source] Simultaneous determination of eight major steroids from Polyporus umbellatus by high-performance liquid chromatography coupled with mass spectrometry detectionsBIOMEDICAL CHROMATOGRAPHY, Issue 2 2010Ying-yong Zhao Abstract Polyporus umbellatus is a widely used diuretic herbal medicine. In this study, a high-performance liquid chromatography coupled with atmospheric pressure chemical ionization,mass spectrometric detection (HPLC-APCI-MS) method was developed for qualitative and quantitative analysis of steroids, as well as for the quality control of Polyporus umbellatus. The selectivity, reproducibility and sensitivity were compared with HPLC with photodiode array detection and evaporative light scattering detection (ELSD). Selective ion monitoring in positive mode was used for qualitative and quantitative analysis of eight major components and ,-ecdysterone was used as the internal standard. Limits of detection and quantification fell in the ranges 7,21 and 18,63 ng/mL for the eight analytes with an injection of 10 µL samples, and all calibration curves showed good linear regression (r2 > 0.9919) within the test range. The quantitative results demonstrated that samples from different localities showed different qualities. Advantages, in comparison with conventional HPLC,diode array detection and HPLC-ELSD, are that reliable identification of target compounds could be achieved by accurate mass measurements along with characteristic retention time, and the great enhancement in selectivity and sensitivity allows identification and quantification of low levels of constituents in complex Polyporus umbellatus matrixes. Copyright © 2009 John Wiley & Sons, Ltd. [source] Determination of total retronecine esters-type hepatotoxic pyrrolizidine alkaloids in plant materials by pre-column derivatization high-performance liquid chromatographyBIOMEDICAL CHROMATOGRAPHY, Issue 6 2009Ai-zhen Xiong Abstract A pre-column derivatization high-performance liquid chromatography method with diode array detection was developed and validated to determine the total retronecine esters-type hepatotoxic pyrrolizidine alkaloids (RET-HPAs) in herbs. The RET-HPAs reacted with o -chloranil in methanolic solution heated for 3 h, and an oxidative derivative was produced that could be detected at a maximal absorption of 223 nm. The analysis was performed using a C18 column with an isocratic elution of methanol and aqueous 0.01% triethylamine (adjusted to pH 4 with formic acid), and the detection was carried out with DAD at 223 nm. The validation of the method included linearity, sensitivity, recovery and stability. It showed a good linear regression (r2 > 0.9900) in the range of 2.5,250 µm with a limit of detection (S/N = 3) of 0.5 µm. The method provided desirable repeatability with overall intra- and inter-day variations of less than 4.6%. The obtained recoveries for both of the extraction and derivatization process were between 94.6 and 100.7% (n = 3). Copyright © 2009 John Wiley & Sons, Ltd. [source] Preparative isolation and purification of alkannin/shikonin derivatives from natural products by high-speed counter-current chromatography,BIOMEDICAL CHROMATOGRAPHY, Issue 2 2009Andreana N. Assimopoulou Abstract Alkannin and shikonin (A/S) and their derivatives have been found in the roots of several Boraginaceous species and are also produced through plant tissue cultures. The chiral compounds A/S are potent pharmaceutical substances with a wide spectrum of biological and pharmacological activities like wound healing, antimicrobial, anti-inflammatory, anticancer and antioxidant activity. High-speed counter-current chromatography (HSCCC) was applied for the first time to the separation, preparative isolation and purification of A/S and their esters from extracts of Alkanna tinctoria roots, as well as commercial samples. The constituents of HSCCC fractions and their purity were determined by high-performance liquid chromatography,diode array detection,mass spectrometry (HPLC-DAD-MS), since DAD cannot detect oligomeric A/S derivatives that are present in most of the samples containing the respective monomeric derivatives. The purity of HSCCC fractions was compared with the one of fractions isolated by column chromatography (CC) using as stationary phases silica gel and Sephadex LH-20. As shown, the purity of monomeric alkannin/shikonin was greater by HSCCC than CC separation of commercial A/S samples. Copyright © 2008 John Wiley & Sons, Ltd. [source] Simultaneous determination of ten amphetamine designer drugs in human whole blood by capillary electrophoresis with diode array detectionBIOMEDICAL CHROMATOGRAPHY, Issue 10 2005Maria Nieddu Abstract In recent years, a number of new designer drugs have entered the illicit drug market. The methylenedioxyderivatives of amphetamine represent the largest group of designer drugs. This paper describes a method for screening for and simultaneously quantifying 10 2,5-methylenedioxy-derivatives of amphetamine and phenethylamine in human whole blood, using capillary electrophoresis (CE) with diode array detection (DAD). Using an aqueous pH 2.5 phosphate buffer, CE analysis gave peaks with good symmetry and reproducible migration times. Under these experimental conditions, the 10 amphetamines were resolved in 15 min and without interference from biological matrices (blood). Their identification by migration time was confirmed by their UV spectra recorded with a DAD (190,350 nm). The main advantages of the present method lie in its simplicity, clean and reliable extraction from human whole blood and simultaneous detection and quantification by CE-DAD. The applicability of the method was demonstrated by analysis of in vivo rat blood samples. The method was validated according to international guidelines. Copyright © 2005 John Wiley & Sons, Ltd. [source] Photohydroxylation of 1,4-Benzoquinone in Aqueous Solution RevisitedCHEMISTRY - A EUROPEAN JOURNAL, Issue 2 2004Justus von Sonntag Dr. Abstract In water, photolysis of 1,4-benzoquinone, Q gives rise to equal amounts of 2-hydroxy-1,4-benzoquinone HOQ and hydroquinone QH2 which are formed with a quantum yield of ,=0.42, independent of pH and Q concentration. By contrast, the rate of decay of the triplet (,max=282 and ,410 nm) which is the precursor of these products increases nonlinearly (k=(2,3.8)×106 s,1) with increasing Q concentration ((0.2,10) mM). The free-radical yield detected by laser flash photolysis after the decay of the triplet also increases with increasing Q concentration but follows a different functional form. These observations are explained by a rapid equilibrium of a monomeric triplet Q* and an exciplex Q2* (K=5500±1000,M,1). While Q* adds water and subsequent enolizes into 1,2,4-trihydroxybenzene Ph(OH)3, Q2* decays by electron transfer and water addition yielding benzosemiquinone .QH and . OH adduct radicals .QOH. The latter enolizes to the 2-hydroxy-1,4-semiquinone radical .Q(OH)H within the time scale of the triplet decay and is subsequently rapidly (microsecond time scale) oxidized by Q to HOQ with the concomitant formation of .QH. On the post-millisecond time scale, that is, when .QH has decayed, Ph(OH)3 is oxidized by Q yielding HOQ and QH2 as followed by laser flash photolysis with diode array detection. The rate of this pH- and Q concentration-dependent reaction was independently determined by stopped-flow. This shows that there are two pathways to photohydroxylation; a free-radical pathway at high and a non-radical one at low Q concentration. In agreement with this, the yield of Ph(OH)3 is most pronounced at low Q concentration. In the presence of phosphate buffer, Q* reacts with H2PO4, giving rise to an adduct which is subsequently oxidized by Q to 2-phosphato-1,4-benzoquinone QP. The current view that . OH is an intermediate in the photohydroxylation of Q has been overturned. This view had been based on the observation of the . OH adduct of DMPO when Q is photolyzed in the presence of this spin trap. It is now shown that Q*/Q2* oxidizes DMPO (k ,1×108,M,1,s,1) to its radical cation which subsequently reacts with water. Q*/Q2* react with alcohols by H abstraction (rates in units of M,1,s,1): methanol (4.2×107), ethanol (6.7×107), 2-propanol (13×107) and tertiary butyl alcohol (,0.2×107). DMSO (2.7×109) and O2 (,2×109) act as physical quenchers. [source] | |||||||||||||