高效液相色谱-二极管阵列检测法测定小麦粉及面粉改良剂中福美双

福美双是重要的二硫代氨基甲酸酯(DTC)杀菌剂,在小麦中使用限量以1 mg/kg二硫化碳(CS2)计.目前我国相关检测方法是针对二硫代氨基甲酸酯一类的化合物,二硫代氨基甲酸酯通过与酸反应生成CS2,采用光谱法或色谱法测定CS2,间接实现二硫代氨基甲酸酯测定.该方法无法特异性实现对福美双的检测,因此开展小麦粉中福美双检测...     

高效液相色谱-二极管阵列检测法及高效液相色谱-电喷雾串联质谱法测定植物源性蛋白中残留的三聚氰胺

建立了高效液相色谱-二极管阵列检测器(HPLC-DAD)及HPLC-电喷雾串联质谱(ESI-MS/MS)测定植物源性蛋白中残留的三聚氰胺的方法。利用HPLC-DAD进行样品中三聚氰胺的初筛,利用HPLC-MS/MS进行确证。采用三氯乙酸溶液沉淀样品中的蛋白,同时提取目标分析物,质谱检测时样品再经强阳离子固相萃取柱富集净化。HPLC-DAD的检测低限为10 mg/kg,HPLC-MS/MS的检测低限为0.5 mg/kg;HPLC-DA的添加回收率为76%～88%,HPLC-MS/MS的添加回收率为72%～82%(基质匹配曲线校正),两种方法的添加回收率的相对标准偏差(RSD)为3.4%～6.4%。     

Improved high-performance liquid chromatography-diode array detection method for the determination of phenolic compounds in leaves and peels from different apple varieties

Hydroxycinnamic acids derivatives, monomeric and oligomeric flavan-3-ols, flavonols, and dihydrocalcones are four of the major polyphenolic groups found in apples leaves and peels. A simple extraction with minimal pre-treatment and a high-performance liquid chromatography-diode array detection determination are optimized and validated, in order to identify and quantitate the polyphenolic profile of leaves and peels of four apples varieties (Gala, Topaz, Golden Delicious, and Florina). The improved chromatographic method has led to better separation of some known polyphenols in a single course, and diode-array detection has been used for the previsional identification of some polyphenolic compounds not available as standards. Because the mobile phase and the chromatographic column are compatible with a mass spectrometer, this method could investigate the unknown flavanols, flavonols, hydrocinnamic acid derivatives, and chalcone-related compounds found in apple leaves and peel extracts analyzed.     

Peak purity determination with principal component analysis of high-performance liquid chromatography-diode array detection data

A method is proposed for the determination of chromatographic peak purity by means of principal component analysis (PCA) of high-performance liquid chromatography with diode array detection (HPLC-DAD) data. The method is exemplified with analysis of binary mixtures of lidocaine and prilocaine with different levels of separation. Lidocaine and prilocaine have very similar spectra and the chromatograms used had substantial peak overlap. The samples analysed contained a constant amount of lidocaine and a minor amount of prilocaine (0.02-2 conc.%) and hence the focus was on determining the purity of the lidocaine peak in the presence of much smaller levels of prilocaine. The peak purity determination was made by examination of relative observation residuals, scores and loadings from the PCA decomposition of DAD data over a chromatographic peak. As a reference method, the functions for peak purity analysis in the chromatographic data system used (Chromeleon) were applied. The PCA method showed good results at the same level as the detection limit of baseline-separated prilocaine, outperforming the methods in Chromeleon by a factor of ten. There is a discussion of the interpretation of the result, with some comparisons with evolving factor analysis (EFA). The main advantage of the PCA method for determination of peak purity over methods like EFA lies in its simplicity, short time of calculation and ease of use.     

Determination of two antifouling booster biocides and their degradation products in marine sediments by high performance liquid chromatography-diode array detection

A method for the simultaneous determination of two antifouling booster biocides, diuron [1-(3,4-dichlorophenyl)-3,3-dimethylurea] and irgarol 1051 (2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine) and their degradation products, DCPMU [1-(3,4-dichlorophenyl)-3-methylurea], DCPU [1-(3,4-dichlorophenyl) urea], DCA [3,4-dichloroaniline] and M1 (2-methylthio-4-tert-butylamino-s-triazine), in marine sediments by high performance liquid chromatography-diode array detection (HPLC-DAD) was developed. The extraction of these compounds from sediment was achieved by means of methanolic ultrasonic extraction. The optimization of the extraction procedure included the variation of the volume of the extraction solvent, the amount of the extracted sediment, the duration and the temperature of sonication. C18 solid phase extraction (SPE) cartridges were used for the cleaning of the extracts. The relative standard deviations (R.S.Ds) of the developed procedure was <10% for all the tested compounds, except for DCA, where R.S.Ds up to 15% were obtained. Satisfactory recoveries were obtained (higher than 80% for all substances), except for DCA, which gave a recovery in the range of 35-50%. The limits of detection (LODs) of the substances varied between 1.7 (DCPU) and 4.0 (DCPMU) ng g⁻¹ of dry sediment.     

Development and validation of stability indicating method for the determination of exemestane by reverse phase high performance liquid chromatography

Exemestane is an aromatase inhibitor used in the treatment of breast cancer. A selective stability-indicating reversed-phase high performance liquid chromatography (RP-HPLC) method has been developed which can separate and accurately quantitate low levels of exemestane. The stability-indicating capability of the method was demonstrated by adequate separation of exemestane and all the degradation product peaks from exemestane peak and also from each other in stability samples of exemestane. Chromatographic separation of exemestane and its degraded products were achieved by using isocratic elution at a flow rate of 1.0 mL/min on a C18 reverse phase column (Phenomenex, size: 250 × 4.60 mm, particle size 5 μm) at ambient temperature. The mobile phase used for the analysis was acetonitrile-water (60:40, %v/v) with UV visible detection at 242 nm. The proposed method was used to study the degradation behavior of drug under various stress conditions as per ICH recommended guidelines.     

Development and validation of a high performance liquid chromatographic method for the simultaneous determination of potassium clavulanate and cefadroxil in synthetically prepared tablets

A simple, sensitive and rapid high performance liquid chromatographic method was developed and validated for the simultaneous determination of potassium clavulanate and cefadroxil in synthetically prepared tablets. Chromatographic separation and detection was carried out on a C-18 column using 0.05 M potassium dihydrogen phosphate buffer (pH 5.0) and acetonitrile in the ratio of 94: 06 (v/v) as mobile phase at wavelength of 225 nm. The method was linear in the concentration range of 3.75–22.5 μg/mL for potassium clavulanate and 15–90 μg/mL for cefadroxil. The flow rate was 1.0 mL/min and the total analysis time was less than 10 min. The mean recoveries was found to be greater than 99% with RSD less than 1.0%. The proposed method was validated by performing linearity, recovery, specificity, robustness, LOD/LOQ and within-day and between-day precision. The chromatographic results obtained from the synthetically prepared tablets show that the method is highly precise and accurate for the simultaneous quantitation of clavulanate potassium and cefadroxil.     

Development and validation of a simple stability-indicating high performance liquid chromatographic method for the determination of miconazole nitrate in bulk and cream formulations

A simple and stability-indicating high performance liquid chromatographic method was developed and validated for the determination of miconazole nitrate in bulk and cream preparations. The extraction step for cream samples consisted in a warming, cooling and centrifugation procedure that assures the elimination of the lipophilic matrix component, in order to avoid further precipitation in the chromatographic system. Separation was achieved on a ZORBAX Eclipse XDB - C18 (4.6 mm × 150 mm, 5 μm particle size) column, using a mobile phase consisting of water, methanol and acetonitrile, in a flow and solvent gradient elution for 15 min. The column was maintained at 25 °C and 10 μL of solutions were injected. UV detection was performed at 232 nm, although employment of a diode array detector allowed selectivity confirmation by peak purity evaluation. The method was validated reaching satisfactory results for selectivity, precision and accuracy. Degradation products in naturally aged samples could be simultaneously evaluated, without interferences in the quantitative analysis.     

Development of a method for the determination of vardenafil in human plasma by high performance liquid chromatography with UV detection

A simple high‐performance liquid chromatographic (HPLC) method with photometric detection is described for the determination of vardenafil hydrochloride, a phosphodiesterase V inhibitor, in human plasma. Chromatographic separation of the analyte and internal standard was achieved on an analytical 250 × 4.6 mm i.d. reversed‐phase Kromasil KR 100 C₁₈ (5 µm particle size) column using a mobile phase of acetonitrile–potassium dihydrogen phosphate (30:70 v/v). The run time was less than 15 min. Column eluate was monitored at 230 nm. The linearity over the concentration range of 10–1500 ng/mL for vardenafil was obtained and the limit of quantification (LOQ) was 10 ng/mL. The method has been applied to analysis of the vardenafil concentrations for application in pharmacokinetic studies. Copyright © 2009 John Wiley & Sons, Ltd.     

Dispersive liquid-liquid microextraction coupled with high-performance liquid chromatography-diode array detection for the determination of N-methyl carbamate pesticides in vegetables

This paper described a simple, rapid and efficient method for the determination of N-methyl carbamate pesticides in tomato, cucumber, carrot and lettuce samples by dispersive liquid-liquid microextraction coupled with HPLC-diode array detection. Some experimental parameters that influenced the extraction efficiency, such as types and volumes of extraction and disperser solvents, extraction time and salt effect were examined and optimized. Under optimum conditions, the LOD of the method were 0.5-3.0 μg/kg depending on the compounds and the kind of vegetables. The linearities of the method were obtained in the range of 10.0-300 μg/kg for aldicarb, MTMC, carbofuran and carbaryl, and 20.0-600 μg/kg for isoprocarb, with the correlation coefficients ranging from 0.9921 to 0.9993. The RSD varied from 2.9 to 7.5% (n=5). The recoveries of the method for the five carbamates from vegetable samples at two different spiking levels were ranged from 77.8 to 98.2%. Results showed that the method we proposed can meet the requirements for the determination of N-methyl carbamate in vegetable samples and was finally applied to the analysis of target pesticides in vegetable samples taken from local markets.     

Chemometric assisted solid-phase microextraction for the determination of anti-inflammatory and antiepileptic drugs in river water by liquid chromatography-diode array detection

In the present work, an analytical method for the simultaneous determination of seven non steroidal anti-inflammatory drugs (naproxen, ketoprofen, diclofenac, piroxicam, indomethacin, sulindac and diflunisal) and the anticonvulsant carbamazepine is reported. The method involves preconcentration and clean-up by solid-phase microextraction using polydimethylsiloxane/divinylbenzene fibers, followed by liquid chromatography with diode array detection analysis. Parameters that affect the efficiency of the solid-phase microextraction step such as soaking solvent, soaking period, desorption period, stirring rate, extraction time, sample pH, ionic strength, organic solvent and temperature were investigated using a Plackett-Burman screening design. Then, the factors presenting significant positive effects on the analytical response (soaking period, stirring rate, stirring time) were considered in a further central composite design to optimize the operational conditions for the solid phase microextraction procedure. Additionally, multiple response simultaneous optimization by using the desirability function was used to find the optimum experimental conditions for the on-line solid-phase microextraction of analytes in river water samples coupled to liquid chromatography and diode array detection. The best results were obtained using a soaking period of 5 min, stirring rate of 1400 rpm and stirring time of 44 min. The use of solid-phase microextraction technique avoided matrix effect and allowed to quantify the analytes in river water samples by using Milli-Q based calibration graphs. Recoveries ranging from 71.6% to 122.8% for all pharmaceuticals proved the accuracy of the proposed method in river water samples. Method detection limits were in the range of 0.5-3.0 microgL(-1) and limits of quantitation (LOQs) were between 1.0 and 4.0 microgL(-1) for pharmaceutical compounds in river water samples. The expanded uncertainty associated to the measurement of the concentration ranged between 8.5% and 29.0% for 20 microgL(-1) of each analyte and between 9.0% and 29.5% for the average of different concentration levels. The main source of uncertainty was the calibration step in both cases.     

One single standard substance for the determination of multiple anthraquinone derivatives in rhubarb using high-performance liquid chromatography-diode array detection

Due to the complexity of the chemical constitution of traditional Chinese medicines (TCMs), now there is a trend to establish methods, using multi-components analysis, for the effective quality control of TCMs. However, the limited availability of multiple reference substances hinders the wide popularization for routine quality control of TCMs. Using an easily available single component contained in the analyte as an internal standard to determine multiple analogues should be a practical option. In this study, we selected rhubarb rhizome as an example, and used emodin, the easily available active component, as the external standard to determine directly the content of emodin in rhubarb, and the same component as the internal standard to simultaneously determine the other six anthraquinones in rhubarb. Compared with the results obtained using the external standard method, this alternative method was found to have no statistically significant differences in our laboratory as verified by the F-test (p = 95%, n = 6). However, due to weak robustness caused by the fluctuation of relative response factors in different laboratories, such a method can only serve as a practical measure to solve the lack of so many chemical standard substances when relative response factors have been determined. This alternative method can then be used without reference substances. Once the corresponding chemical standards are available and are acceptable as well as cost-effective, then the external standard method for the simultaneous determination of multiple components (SDMC) in TCMs will prevail.     

Simultaneous determination of 3,4-dimethoxybenzaldehyde and 3,4-dimethoxyphenylacetone in industrial waste waters by high-performance liquid chromatography-diode array detection

The environmental pollutants 3,4-dimethoxybenzaldehyde (DMB) and 3,4-dimethoxyphenylacetone (DMPA) were separated and quantitatively determined in treated and untreated industrial waste waters on a porous graphitized carbon column using HPLC with diode array detection (DAD). It was established that the detector response is linear in a wide range of injected quantities for both pollutants, and the logarithm of the capacity factor depends linearly on the concentration of acetonitrile in the eluent. The relative standard deviation of the retention time, peak height and peak area was lower than 1% in the normal and lower than 3% in the lowest concentration range. Peak purity tests indicated that the method separates well DMB and DMPA from other impurities present in the waste waters. Both aerobic and anaerobic treatments markedly decreased the concentration of DMB and DMPA in industrial waters and HPLC combined with DAD proved to be an adequate analytical procedure to follow such changes.     

Chemometric analysis of high performance liquid chromatography-diode array detection-electrospray mass spectrometry of 2- and 3-hydroxypyridine

Triply coupled high performance liquid chromatography using diode array detection and positive ion electrospray mass spectrometry of 2- and 3-hydroxypyridine is presented. Considerations of the physical method for coupling the two detectors, the influence of pH on retention times, the cone voltage of the mass spectrometer and the linear concentration ranges are described. Data from both detectors are aligned and interpolated. The analyte mass spectra are reduced to 20 significant masses. Principal components plots on the raw, normalised and standardised data, derivatives to determine composition 1 regions, deconvolution and procrustes analysis to compare data from both detectors are discussed. Common trends in both mass spectral and diode array chromatograms are interpreted. This paper represents a new approach to common processing of chromatographic data from two detectors.     

Multiresidue method for fourteen fungicides in white grapes by liquid-liquid and solid-phase extraction followed by liquid chromatography-diode array detection

A quantitative, selective and sensitive HPLC method for the analysis of 14 fungicides in white grapes for vinification is described. The proposed method is based on liquid-liquid extraction (LLE) and solid-phase extraction (SPE) followed by liquid chromatography and diode array detection (HPLC-DAD). Dichloromethane-acetone (75:25, v/v) was the most appropriate solvent mix for extracting fungicides in white grapes. Silica cartridges resulted the most appropriate for extract purification purposes. Quality parameters of the proposed multiresidue method presented good recovery (ca. 85% for almost all target compounds) and precision (between 1.5 and 16%), and detection limits lower than maxima residual limits set by the 76/895/ECC and 90/642/ECC Directive. Five different white grapes for vinification produced in Rias Baixas area in Galicia (NW Spain) were analyzed in order to assess the performance of the method with real samples and to determine whether the concentration of the pesticides used exceed their maxima residue levels (MRLs). Results showed that grape concentrations for those identified fungicides were lower than those established by European legislation.     

Determination of porphyrins and metalloporphyrins using liquid chromatography-diode array detection and mass spectrometry

A HPLC procedure has been optimized and successfully applied to porphyrins of environmental interest, such as etio and octaethylporphyrins and their VO and Ni compounds. The use of UV-Vis diode array allowed the detection of the analytes within the 5-15 ng/ml range. In order to achieve greater specificity and some structural information, the coupling of liquid chromatography with mass spectrometry was investigated, and the particle beam interface conditions were optimized. Electron impact (EI) spectra, comparable to those reported in the literature were obtained. The entire procedure has been applied to a real marine sediment, previously spiked with porphyrins to resemble oil-contaminated samples. The results pointed out that the method is suitable for such levels of analytes (5-10 microg/ml), allowing their identification and quantification with no need for purification steps.     

Dispersive liquid-liquid microextraction followed by high-performance liquid chromatography-diode array detection as an efficient and sensitive technique for determination of antioxidants

Dispersive liquid-liquid microextraction (DLLME) and high performance liquid chromatography-diode array detection (HPLC-DAD) was presented for extraction and determination of Irganox 1010, Irganox 1076 and Irgafos 168 (antioxidants) in aqueous samples. Carbon tetrachloride at microliter volume level and acetonitrile were used as extraction and dispersive solvents, respectively. The main advantages of method are high speed, high enrichment factor, high recovery, good repeatability and extraction solvent volume at μL level. Limit of detection for analytes is between 3 and 7 ng mL⁻¹. One variable at a time optimization and response surface modeling were used to obtain optimum conditions for microextraction procedure and nearly same experimental conditions were obtained using both optimization methods. Recoveries in the ranges 78-86% and 84-110% were obtained by one variable at a time and response surface modeling, respectively. Using tap water and packed water as matrices do not show any detrimental effect on the extraction recoveries and enrichment factors of analytes.     

Development and validation of a solid-phase extraction method coupled to high-performance liquid chromatography with ultraviolet-diode array detection for the determination of sulfonylurea herbicide residues in bovine milk samples

This study proposes a fast, simple and sensitive liquid chromatography diode array detector (LC/UV-DAD)-based method for the simultaneous determination of eight sulfonylurea herbicides (bensulfuron methyl, chlorsulfuron, metsulfuron methyl, primisulfuron methyl, rimsulfuron, thifensulfuron methyl, triasulfuron and tribenuron methyl) in bovine whole milk at concentrations lower than the default limit of 0.01 mg kg(-1) allowed by current legislation (Regulation EC/396/2005 and following Annexes). An effective one-step solid phase extraction (SPE) and clean up procedure was defined with use of Chem Elut cartridges, providing good recoveries for all the analytes tested and with no matrix effects affecting method accuracy. Separation of herbicides was obtained on a C(18) column by acetonitrile- water gradient elution. Method validation has been performed according to European Commission Decision 2002/657/EC criteria, in terms of linearity, recovery, precision, specificity, decision limit (CC(α)) and detection capability (CC(β)). Typical recoveries ranged between 78.4% and 99.7%, at the maximum residue limits (MRLs) levels established by Regulation EC/396/2005, with relative standard deviations (RSD) no larger than 10%.     

Simultaneous determination of carbonyl compounds and polycyclic aromatic hydrocarbons in atmospheric particulate matter by liquid chromatography-diode array detection-fluorescence detection

Simultaneous analysis of 24 carbonyl compounds (alkanals, unsaturated, dicarbonylic and aromatic aldehydes and ketones) derivatized with 2,4-dinitrophenylhydrazine and 16 polycyclic aromatic hydrocarbons (PAHs) using a photodiode-array (PDA) and a fluorescence (FL) detector in series is proposed.The separation is carried out with a reversed-phase column and gradient elution using four solvents (acetonitrile, water, tetrahydrofuran and methanol) in less than 35 min. Several critical pairs of carbonyl compounds with 3 and 4 carbon atoms and different functional groups, isomers of tolualdehyde, aromatic and aliphatic aldehydes were conditional on the gradient elution. Common pre-treatment for two groups of compounds consists in a step of extraction and derivatization in aqueous medium and a further clean-up using a polymeric phase SPE and concentration in a mixture of dichloromethane:methanol. A pre-concentration factor of 50 was achieved by this procedure. Acetone and formaldehyde blanks were minimized and remain controlled with a specific cleaning of glass material and washing the SPE cartridge.The limits of detection (LOD) ranged from 0.006 to 0.18 ng mL⁻¹ for PAHs and from 2.4 to 10.1 ng mL⁻¹ for carbonyl compounds and method precision was ≤15% for all analysed compounds. Recoveries were within the range of 95-104% for PAHs except for more volatile compounds (acenaphthene and fluorene) and within the range of 72-113% for carbonyl compounds. The method was applied in water-soluble fraction of PM₁₀ (atmospheric particulate matter with an aerodynamic diameter less than 10 μm) and the spectral contrast technique was used in the identification of carbonyl compounds.