HPLC-integrated solid-phase extraction with photochemical post-column derivatization for the determination of oxacillin, cloxacillin and dicloxacillin in raw milk

 An automated reversed-phase high-performance liquid chromatography (HPLC) method with UV detection at 300 nm after on-line coupled solid-phase extraction (SPE) and photochemical post-column derivatization was developed for the residue analysis of oxacillin, cloxacillin and dicloxacillin in raw milk. After a centrifugation step for defatting and ultrafiltration, a HPLC-integrated SPE using a restricted access sorbent, is performed. By operating the system in parallel mode, SPE and HPLC run are performed simultaneously. The procedure allows the analysis of about 30 samples within 24 h and was used to investigate the milk of cows which had been treated with oxacillin. The detection limit was 3 μg/kg for oxacillin and cloxacillin and 5 μg/kg for dicloxacillin. The recoveries were between 66% and 81% with coefficients of variation between 3.3% and 7.7%. Using an acetonitrile extraction procedure better recoveries of ≥90% were obtained with fortified samples, but more or less identical results were obtained for the samples with incurred residues after correction for recovery. Received: 16 February 1998     

Analytical methods for the determination of pesticide residues using gas chromatograghy with nitrogen-phosphorus detector

The analysis methods for 3 pesticides (ametryn, cyanophos, and fonofos) were developed and validated. One 1,3,5-triazine herbicide (ametryn), and 2 organophosphorus insecticides (cyanophos and fonofos) were tested. Three kinds of residual pesticides were classified into 2 groups according to the chemical structure and separation method. Solid-phase extraction (SPE) cartridge was used for sample purification and isolation of analytes. Final solution was analyzed with GC with nitrogen-phosphorus detection (NPD). Confirmation analysis of pesticides was carried out by GC-MS in the selected ion monitoring (SIM) mode. Correlation coefficient was ranged from 0.999 to 1.0, depending on sample analytes. Rice, apple, and soybean samples were selected for recovery experiment. Overall recovery rates from blank samples spiked at 3 fortification levels ranged from 76.29 to 107.4% with relative standard deviations lower than 20.2%. The limit of quantification was <3–5 μg/L depending on the analytes, and the reporting level of the method, defined as lower than the maximum residue levels established by Korea Food & Drug Administration (KFDA).     

Simultaneous Determination of 69 Pesticide Residues in Coffee by Gas Chromatography?CMass Spectrometry

A method using gel permeation chromatography (GPC) combined with solid-phase extraction (SPE) cleanup followed by gas chromatography–mass spectrometry (GC-MS) has been established for quantitative determination of 69 pesticide residues in coffee. Based on an appraisal of the characteristics of GC-MS, validation experiments were conducted for 69 pesticides. In the method, 2.0 g samples were mixed with 5 ml water and 1 g sodium chloride and extracted with 5 ml of ethyl acetate by blender homogenization, centrifugation, and filtration. Evaporation was conducted and the sample was injected into a 250 mm × 10 mm S-X3 GPC column, with ethyl acetate–n-hexane (1:2 v/v) as the mobile phase at a flow rate of 3 ml/min. The 4–15 min fraction was collected for the SPE cleanup, which was Envi-Carb SPE cartridge coupled with NH₂-LC SPE cartridge with acetone–ethyl acetate (2:5 v/v) as the eluted solvent. The eluents were collected and then evaporated to dryness, which was redissolved in 0.5 ml ethyl acetate for GC-MS analysis. For the 69 pesticides determined by GC-MS, the portions collected from GPC were concentrated to 0.5 ml and exchanged with 5 ml n-hexane. In the linear range of each pesticide, the correlation coefficient was R ² ≥ 0.99. At the low, medium, and high fortification levels of 0.05–1.0 mg/kg, recoveries fell within 60–120%. The relative standard deviation was between 1.3% and 22.3% for all 69 pesticides. The limits of detection for the method were 10 μg/kg to 150 μg/kg, depending on each pesticide.     

SPE and MSPD as pre-separation techniques for HPLC of tetracyclines in meat, milk and cheese

 Solid phase extraction (SPE) and Matrix solid phase dispersion (MSPD) have been tested as pre-separation procedures for high-performance liquid chromatography (HPLC) determination of tetracycline antibiotics in milk, meat and cheese. The extraction recoveries ranged from 48% to 86% for SPE and from 89% to 93% for MSPD at concentration levels of the maximal residual limits (MRL) recommended by the European Union to be 100 ng/g for the tetracyclines oxytetracycline (OTC), tetracycline (TC) and chlortetracycline (CTC). The detection limits were 15 – 22 ng/g for SPE and 30 ng/g for MSPD. Following the relatively simple SPE procedure we used a Lichrosorb RP-18 column and a diode array detector for HPLC. Results dealing with analysis of tetracyclines HPLC in cheese are published for the first time. Received: 7 March 1997     

动态微波辅助萃取法快速萃取苹果中8种 氨基甲酸酯类农药

目的 建立一种绿色的动态微波辅助快速萃取法结合高效液相色谱-串联质谱法测定苹果中八种氨基甲酸酯类农药的方法。方法 首先,以水为萃取溶剂,利用动态微波辅助萃取法对苹果中氨基甲酸酯类农药进行快速萃取;之后通过两步固相萃取法对所获得的萃取物进行净化和富集;最后,采用高效液相色谱-串联质谱法对目标化合物进行分析。结果 实验中对影响动态微波辅助萃取和固相萃取效率的实验条件进行了优化,最优的实验条件为：萃取剂为纯水,微波萃取功率为600 W;微波萃取时间为10 min;萃取剂流速为1.0 mL min-1;固相萃取洗脱剂为2 mL的乙醇。八种氨基甲酸酯类农药在8~800 ng g-1范围内成线性关系,相关系数为0.994~0.999;检出限为1.1~4.2 ng g-1;方法的加标(8 ng g-1)回收率为71%~90%。结论 该方法可以用于苹果中氨基甲酸酯类农药的快速检测。     

Analysis of patulin in apple products by liquid–liquid extraction,solid phase extraction and matrix solid-phase dispersion methods: a comparative study

Patulin is a marker of quality in the apple and apple juice industry and due to the potential risk for human health, reliable and potential methods for extracting patulin from a sample are therefore needed. In this study, the three methods with liquid–liquid extraction, matrix solid-phase dispersion (MSPD) and with solid-phase extraction (SPE) were studied for extracting patulin from different apple products. Result showed that for AJC and apple sample, MSPD method is most suitable for extracting patulin among the three methods. The recovery rates of AJC and apple sample were 80.35–114.46 and 79.68–94.32%, respectively, the coefficient variations were 3.18–4.90%; For dilute juice, SPE procedure is suitable for analysis of patulin and the recovery rates were and 85.35–90.14%.     

Analysis of pesticide residues in sugarcane juice using QuEChERS sample preparation and gas chromatography with electron capture detection

QuEChERS sample preparation was used for the determination of 7 pesticides residues in 80 samples of sugarcane juice collected from two Brazilian cities, in two different periods. The method involved extraction with acetonitrile, liquid–liquid partition with addition of MgSO₄ and NaCl followed by dispersive SPE cleanup with PSA sorbent and the analyses were carried out with a GC–ECD equipment. The method was validated using sugarcane juice spiked at 0.025, 0.10 and 0.20 mg/L and the average recovery by the method varied from 62.9% to 107.5% with RSDs < 18%. The method showed good linearity and the LODs for the pesticides studied ranged from 0.003 to 0.04 mg/L. No pesticide residue was detected (>LOD) amongst the 80 samples analysed.     

Solid Phase Extraction Combined Direct Spectrophotometric Determination of Brilliant Blue in Food Using β-Cyclodextrin Polymer

A new and sensitive solid phase extraction (SPE) combined direct spectrophotometry for determination of brilliant blue (BB) in food has been developed, in which β-cyclodextrin polymer (β-CDP) is used as a SPE adsorbent to extract BB from aqueous solutions to form a solid supramolecular complex with a 1:3 molar ratio of BB to β-CDP, and BB in β-CDP was directly determined by a solid phase spectrophotometry. SPE conditions were pH 7.0, temperature 20 °C, ionic strength 0.1 mol/l, and shaking time 40 min. The calibration curve is linearity in the concentration range of 0.05–12.0 μg/ml. The proposed method was applied to the determination of BB in food samples at 627 nm maximum absorption wavelength with satisfactory results.     

A new quick solid-phase extraction method for the quantification of heterocyclic aromatic amines

A new solid-phase extraction (SPE) method using only one SPE cartridge is described as a clean-up procedure for the determination of heterocyclic aromatic amines (HAAs). In particular, the polar HAAs imidazoquinolines and imidazoquinoxalines, which are well-known toxic compounds in thermally treated food, can be determined by this quick and simple method. For validation of the method meat extracts were analyzed. For determination of the percentage recovery and standard deviation the beef extract was spiked (40 ng/g of each HAA) and analyzed 10 times. The polar HAAs were determined in this meat extract with a recoveries of 62 % to 95% and standard deviations of 3% to 5%. The recovery rates of the less polar HAAs Harman and Norharman were much lower (25±5% to 32±5%). The limit of detection for polar and less polar HAAs was between 3 ng/g and 9 ng/g in the meat extract matrix. The method comprises extraction with methanolic NaOH, centrifugation and SPE using a commercially available polystyrene copolymer cartridge. After different washing steps the eluate was analyzed by high-performance liquid chromatography with diode-array detection. The advantages of this new method are the reduced amounts of time and organic solvents required. Received: 29 November 1999 / Revised version: 21 February 2000     

Rapid multiresidue determination of pesticides in livestock muscle and liver tissue via modified QuEChERS sample preparation and LC-MS/MS

ABSTRACT

Many multiresidue methods for the determination of pesticides in vegetables and fruits have been reported to date. However, few such methods have been employed to investigate pesticide residues in animal tissue. In this study, an LC-MS/MS multiresidue method coupled with modified QuEChERS extraction was developed and validated for the investigation of eight pesticide residues: prallethrin (PR), resmethrin (RMT), imidacloprid (IMC), diflubenzuron (DFB), cyromazine (CYR), etofenprox (EFP), dinotefuran (DNT) and phthalthrin (PTLT). This method involves initial extraction in a water/acetone system, the addition of salts and a subsequent extraction/partitioning step and, finally, a clean-up step utilising dispersive solid-phase extraction (SPE). The mean recoveries of seven of the pesticides (the exception being CYR) ranged between 74.7% and 113.5%, and the CVs of the livestock tissue – bovine, swine, and chicken muscle and liver tissue spiked at 10 ng g^–1 (50 ng g^–1 for RMT and DNT) and 100 ng g^–1 – were < 13.8%. The recoveries of CYR in all muscle and liver spiked samples ranged from 56.9% to 78.3%, while those of RMT in swine liver were > 120%. Therefore, this method was considered as being unsuitable for the investigation of these samples. The limits of quantitation (LOQs) of seven of the investigated pesticides (the exception being swine liver) in the tissue samples ranged from 0.9 to 15.2 ng g^–1. We therefore concluded that this LC-MS/MS multiresidue method is a valid and suitable for the investigation of seven pesticides in animal tissue, but it is unsuitable for the analysis of CYR in all animal tissues and RMT in swine liver tissue.     

固相萃取-液相色谱法测定水果中的氨基甲酸酯与有机磷农药

建立了固相萃取-高效液相色谱同时测定水果中4种氨基甲酸酯与一种有机磷的方法.对提取溶剂、固相萃取柱种类、洗脱剂类型和用量及检测的色谱条件进行筛选和优化.样品经乙腈超声提取,弗罗里硅土固相萃取柱净化,净化后的样品采用液相色谱柱分离,以甲醇:水(体积比70∶ 30)为流动相,紫外检测波长为210nm,流速1.0mL/min...     

Concentration dependence of recovery rates of an exogenous short linear DNA fragment from soya flour

Recovery rates of an exogenous short linear DNA fragment from soya flour by two methods for isolation of DNA—cetyltrimethylammonium bromide solubilization with liquid-liquid extraction (CTAB-LLE) and chaotropic solid-phase extraction (SPE)—were determined using quantitative real-time polymerase chain reaction. Recovery rates varied from 3 to 54% for CTAB-LLE and from 8 to 66% for chaotropic SPE in a concentration-dependent manner, the highest values being achieved only at high concentrations of the DNA fragment (400ng(100mg matrix for CTAB-LLE and =100ng(100mg matrix for chaotropic SPE). These results suggest that if an exogenous DNA fragment is intended to be used as an internal standard, its working concentrations have to be selected very carefully. Presented results also underline the fact that recovery rates of the currently used methods for DNA isolation from food are relatively low and represent an important problem in the development of quantitative DNA-based methods for food analysis.     

Gas chromatographic determination of pesticide residues using electron-capture detector and mass spectrometry

The sensitive and reliable method for determination of 6 pesticides, cyanazine (triazine), uniconazole (azole), diflufenican (nicotinilide), vernolate (thiocarbamate), bromoxynil (hydroxyl benzonitrile), and asulam (sulfanilyl carbamate) were examined and validated. The methods based on solid-phase extraction (SPE) and gas chromatograph (GC) with electron capture detection (ECD). Confirmation analysis of pesticides was carried out by GC-MS in the selected ion monitoring (SIM) mode using 2 target ions. Method validation was performed at 5 fortification levels (0.005, 0.01, 0.05, 0.1, and 0.5 mg/L). The linearity of the calibration curves was excellent in matrix matched standards, and yielded the coefficients of determination (R ²)≥0.9982 for all target analytes. The limit of quantification (LOQ) was 0.05 mg/L for 6 pesticide compounds and the reporting level of the method, defined as lower than the maximum residue levels established by Korea Food & Drug Administration (KFDA). Average recoveries of the pesticides spiked at 0.05, 0.1, and 0.5 mg/L into 3 agricultural products, rice, apple, and soybean were in the range 76.40–120% with relative standard deviation (RSD) values ≤11.07% for all compounds, respectively.     

Assessment of pesticide residues in fresh peach samples produced under integrated crop management in an agricultural region of northern Greece

A multi-residue method using selected ion monitoring mode GC/MS has been developed for the quantitative analysis of residue levels of 23 widely used pesticides in fresh peaches produced under integrated crop management process (ICM). The proposed methodology involved a sample extraction procedure using liquid–liquid partition with acetonitrile followed by a clean-up step based on solid-phase extraction (SPE). Fortification studies were performed at different concentration levels for various types of peaches that differ in properties, such as appearance, flavor and pit. The data showed that the different peach matrices had no significant effect on recoveries. Recoveries were greater than 80% for most of the pesticides with a RSD below 18%. The limits of quantification (LOQs) were in the range 0.002–0.050 mg kg^?1, depending on the compound. To assess method performance with real samples and determine whether pesticide concentrations in peaches exceed their maximum residue levels (MRLs), the proposed method was successfully applied to the analysis of 104 fruit samples collected under integrated pest management (IPM) production during the 2006 cultivation period. Residues detected were lower than those established by legislation for all pesticides, except diazinon, where one positive sample was detected at a level of 0.03 mg kg^?1.     

Sample preparation for photometric determination of histamine in foodstuffs compared with capillary electrophoresis

 To determine levels of histamine, two methods were used, photometry in conjunction with two sample clean-up procedures, and capillary zone electrophoresis (CZE). The two sample clean-up procedures used were liquid liquid extraction (LLE) with n-butanol and solid phase extraction (SPE). Using CZE, the separation of histamine from the matrix was good. The other method, photometry, represents a classic and simple method, that can be employed for in situ measurement of histamine. We found that it was necessary to clean up the samples prior to photometry; if this was not done, the recorded levels of histamine were higher than those determined by CZE. In order to determine levels of histamine, both of these rapid tests were applied to ten different foodstuffs. The levels of histamine measured using photometry following either LLE or SPE were compared. The results indicated that photometry is a suitable method for the measurement of histamine, although the sample solutions have to be purified by either LLE or SPE. Samples do not need to be cleaned up before CZE because there is no interference between histamine and attendant material. Both sample clean-up procedures were applied to the following foodstuffs: tomatoes, sauerkraut, tuna, leaf spinach, cream spinach, white wine and mackerel. The differences of the measured values vary between 3% and 18% for LLE and 6% and 27% for SPE. For the other foodstuffs, such as beef, beer and non-alcoholic beer, only one sample clean-up procedure is suitable. LLE used for beef and beer leads to differences in measured levels of histamine between 18% and 50%, respectively, whereas SPE used for non-alcoholic beer leads to differences of 20%. Received: 3 September 1997 / Revised version: 23 October 1997     

Sample preparation for photometric determination of histamine in foodstuffs compared with capillary electrophoresis

 To determine levels of histamine, two methods were used, photometry in conjunction with two sample clean-up procedures, and capillary zone electrophoresis (CZE). The two sample clean-up procedures used were liquid liquid extraction (LLE) with n-butanol and solid phase extraction (SPE). Using CZE, the separation of histamine from the matrix was good. The other method, photometry, represents a classic and simple method, that can be employed for in situ measurement of histamine. We found that it was necessary to clean up the samples prior to photometry; if this was not done, the recorded levels of histamine were higher than those determined by CZE. In order to determine levels of histamine, both of these rapid tests were applied to ten different foodstuffs. The levels of histamine measured using photometry following either LLE or SPE were compared. The results indicated that photometry is a suitable method for the measurement of histamine, although the sample solutions have to be purified by either LLE or SPE. Samples do not need to be cleaned up before CZE because there is no interference between histamine and attendant material. Both sample clean-up procedures were applied to the following foodstuffs: tomatoes, sauerkraut, tuna, leaf spinach, cream spinach, white wine and mackerel. The differences of the measured values vary between 3% and 18% for LLE and 6% and 27% for SPE. For the other foodstuffs, such as beef, beer and non-alcoholic beer, only one sample clean-up procedure is suitable. LLE used for beef and beer leads to differences in measured levels of histamine between 18% and 50%, respectively, whereas SPE used for non-alcoholic beer leads to differences of 20%. Received: 3 September 1997 / Revised version: 23 October 1997     

HPLC determination of stevioside in plant material and food samples

 An HPLC method for the determination of sweet-tasting stevioside (STS) in the leaves of the plant Stevia rebaudiana and in some beverages (e.g. tea, orange juice) was developed. The pre-separation procedure consisted of extraction of STS from the plant material using boiling water and a solid-phase extraction (SPE). Recovery rates of the SPE for the analysed matrices ranged from 92.8% to 97.8% (for concentrations of STS of 105, 210 and 300 μg/ml; Relative Standard Deviation (RSD)≤3.3%). The chromatographic separations were realized using a C₁₈ column, the mobile phase consisting of methanol and water, and with UV detection at 210 nm. The limits of determination of STS were 5 μg/ml for the leaf extracts and the tea sample and 8 μg/ml for the juice sample. Received: 7 April 1998     

HPLC determination of stevioside in plant material and food samples

 An HPLC method for the determination of sweet-tasting stevioside (STS) in the leaves of the plant Stevia rebaudiana and in some beverages (e.g. tea, orange juice) was developed. The pre-separation procedure consisted of extraction of STS from the plant material using boiling water and a solid-phase extraction (SPE). Recovery rates of the SPE for the analysed matrices ranged from 92.8% to 97.8% (for concentrations of STS of 105, 210 and 300 μg/ml; Relative Standard Deviation (RSD)≤3.3%). The chromatographic separations were realized using a C₁₈ column, the mobile phase consisting of methanol and water, and with UV detection at 210 nm. The limits of determination of STS were 5 μg/ml for the leaf extracts and the tea sample and 8 μg/ml for the juice sample. Received: 7 April 1998     

Comparison of Simple and Rapid Extraction Procedures for the Determination of Pesticide Residues in Fruit Juices by Fast Gas Chromatography–Mass Spectrometry

Three sample treatment methods, based on QuEChERS, solid-phase extraction (SPE) and solid-phase microextraction (SPME), were compared and evaluated in order to obtain the best conditions to determine pesticide residues in fruit juice by fast gas chromatography–mass spectrometry (single quadrupole GC-MS). Analysis were performed under selected ion monitoring, acquiring the three most abundant and/or specific ions for each analyte and using their relative intensity ratios as a confirmatory parameter. The 3 methodologies (QuEChERS, SPE and SPME) were validated taking 15 selected pesticides as model compounds, using commercial apple juice. QuEChERS procedure was based on the AOAC Official Method 2007.01, using acetonitrile (containing 1 % acetic acid) as extraction solvent and primary–secondary amine during the dispersive solid-phase extraction. Oasis hydrophilic–lipophilic balance cartridges were used for SPE, and polyacrylate fibers were used for direct immersion SPME procedure. Three isotopically labeled standards were added to the samples before extraction and used as surrogate standards. Validation parameters as recoveries, limits of detection, and limits of quantification (LOQ), as well as matrix effects and sample throughput, were obtained and compared for the three extraction procedures. QuEChERS was considered faster and led to the best quantitative results. In this way, validation was extended to up to 56 pesticides by applying QuEChERS in multi-fruit juice samples, obtaining LOQs ranging from 2 to 20 μg/L for most compounds. Accuracy and precision were evaluated by means of recovery experiments at two concentration levels (10 and 100 μg/L), obtaining recoveries between 70 and 120 % in most cases and relative standard deviations below 15 %. Finally, the QuEChERS method was applied to the analysis of commercial juices, including mango–apple, pineapple, grapefruit and orange.     

Determination of ochratoxin A in wheat after clean-up through a DNA aptamer-based solid phase extraction column

A DNA aptamer with high affinity and specificity to ochratoxin A (OTA) was conjugated to a coupling gel and used as sorbent for the preparation of solid phase extraction (SPE) columns. The SPE columns packed with 300 μl oligosorbent (24 nmol DNA) showed a linear (r = 0.999) behaviour in the range of 0.4–500 ng OTA. After optimisation of the extraction step, SPE columns were used for clean-up of OTA from wheat prior to liquid chromatographic (HPLC) analysis with fluorescence detection (FLD). Average recoveries from wheat samples spiked at levels of 0.5–50 ng/g ranged from 74% to 88% (relative standard deviation <6%) with limits of detection and of quantification of 23 and 77 pg/g, respectively. The comparative HPLC/FLD analyses of 33 naturally contaminated durum wheat samples cleaned-up on both aptamer-SPE and immunoaffinity (IMA) columns showed a good correlation (r = 0.990). Aptamer-SPE columns could be re-used up to five times without any loss of performance.