Determination of catechins in green tea infusions by reduced flow micellar electrokinetic chromatography

A sulfated-β-cyclodextrin (s-β-CD) modified reduced flow micellar electrokinetic chromatography (RF-MEKC) method was developed and validated for the determination of catechins in green tea. The optimal electrolyte consisted of 0.2% triethylamine, 50 mmol/L SDS and 0.8% s-β-CD (pH = 2.9), allowing baseline separation of five catechins in 4 min. The samples and standards were injected at 0.6 psi for 5 s under constant voltage of −30 kV. Sample preparation simply involved extraction of 2 g of tea with 200 mL water at 95 °C under constant stirring for 5 min. The method demonstrated excellent performance, with limits of detection (LOD) and quantification (LOQ) of 0.02–0.1 and 0.1–0.5 μg/mL, respectively, and recovery percentages of 94–101%. The method was applied to six samples of Brazilian green tea infusions. Epigallocatechin gallate (23.4–112.4 μg/mL) was the major component, followed by epigallocatechin (18.4–78.9 μg/mL), epicatechin gallate (5.6–29.6 μg/mL), epicatechin (4.6–14.5 μg/mL) and catechin (3.2–8.2 μg/mL).     

Rapid determination of organophosphorous pesticides in leeks by gas chromatography–triple quadrupole mass spectrometry

A rapid multi-residue method was developed for the determination of 20 organophosphorous pesticide residues in leeks by gas chromatography coupled to triple quadrupole mass spectrometry (GC–QqQ-MS/MS). The method was based on the modified QuEChERS sample preparation method. After microwave pre-treatment, leek samples were extracted with acetonitrile containing acetic acid 0.1% and cleaned by dispersive solid phase extraction. The QqQ analyser acquired data in selected reaction monitoring (SRM) mode. Recoveries of 20 organophosphorous pesticides ranged from 81.0% to 109.4% with the relative standard deviations (RSD) below 10.4%. The limits of detection (LODs) were 0.07–1.5 μg/kg. The limits of quantitation (LOQs) ranged from 0.25 to 5 μg/kg. Ten leek samples were analysed for method application.     

A multi-residue method for fast determination of pesticides in tea by ultra performance liquid chromatography–electrospray tandem mass spectrometry combined with modified QuEChERS sample preparation procedure

A multi-residue method was developed for rapid determination of pesticide residues in tea by ultra performance liquid chromatography–electrospray tandem mass spectrometry (UPLC/MS/MS). The QuEChERS method was used for sample preparation. In order to minimise the matrix effects from tea, a solid phase extraction (SPE) cartridge layered with graphite carbon/aminopropylsilanized silica gel was applied as complementary to QuEChERS method. For accurate quantification, representative matrix-matched calibration curves were applied to compensate matrix effects. Limits of quantification varied with different pesticides but all can be measured at 0.01 mg kg⁻¹ level in a 5 g tea sample except dichlorvos (0.02 mg kg⁻¹). Recoveries ranged from 70% to 120% and relative standard deviation (RSD) met the European United Quality Control guideline. Efficiency and reliability of this method were investigated by the analysis of both fermented and unfermented Chinese tea samples.     

Fumonisin level in corn-based food and feed from Linxian County,a high-risk area for esophageal cancer in china

The level of mycotoxin fumonisins in corn-based food and feed collected from Linxian County, a high-risk area for esophageal cancer in China, has been analyzed using high-performance liquid chromatographic coupled with evaporative laser scattering detector (HPLC-ELSD). A total of 104 corn kernel samples were obtained from local households, granaries, wholesale markets (central markets), and retail markets (stores and supermarkets). Fumonisin B₁ (FB₁) was detected in the samples from households, granaries, central markets, and stores, with a positive rate of 61.5%, 50%, 33.3%, and 17%, respectively. No fumonisin was detected in samples from the supermarket. The highest FB₁ levels (0.30–3.20 μg/g; mean, 1.42 μg/g) were found in samples from the granary, followed by household (0.25–1.80 μg/g; mean, 0.73 μg/g), central market (0.25–1.10 μg/g; mean, 0.51 μg/g), and store (0.22–0.34 μg/g; mean, 0.28 μg/g). Among the 80 corn kernel samples collected from local households, 18 of 24 (75.0%) moldy samples contained high levels of FB₁ (0.28–3.30 μg/g; mean, 1.58 μg/g), and 20 of 56 (35.7%) apparently healthy samples contained low levels of FB₁ (0.21–0.82 μg/g; mean, 0.46 μg/g). As the central market plays an important role in trade of corn-based food and feed in China, a total of 115 corn-based food and feed samples were collected from the local central market. The highest FB₁ levels (0.30–3.13 μg/g; mean, 1.50 μg/g) were found in feed, followed by unprocessed food (0.31–0.63 μg/g; mean, 0.47 μg/g) and processed food (0.21–0.28 μg/g; mean, 0.25 μg/g). The positive incidence of FB₁ in feed, unprocessed, and processed food were 53.6%, 33.3% and 17.9%, respectively. In conclusion, the results showed that corn-based food and feed from Linxian County contained low level of FB₁ (<2 μg/g) in general, but efforts should be made to control the fumonisin contamination in corn kernels stored in granaries and households.     

Comparison of positive and negative ion detection of tea catechins using tandem mass spectrometry and ultra high performance liquid chromatography

Tea catechins are an important group of natural compounds associated with health promoting effects and desired commodities for the growing market of dietary supplements and functional foods. Consequently these compounds attract more interest of research groups worldwide. A reliable quantitative analysis of tea catechins is essential for human intervention studies, manufacturers of dietary supplements and quality control by authorities. UHPLC–ESI-MS/MS analytical method was chosen due to rapid runtime, high sensitivity and selectivity. The chromatographic separation of eight tea catechins was achieved within 2.5 min on C₁₈ BEH analytical column (100 mm × 2.1 mm i.d.; 1.7 μm), whilst the gradient elution mode was employed using water:methanol mobile phase with addition of volatile organic acid. The concentration of organic acids in the mobile phase was optimised within the range of 0.01–0.1% (v/v). High sensitivities were achieved in positive (10.2-16.8 fmol/inj.) and negative ion detection mode (102.1-168.1 fmol/inj.), through accurate and complex tuning of MS parameters. The UHPLC-ESI-MS/MS method was validated in terms of linearity (>0.9997; >0.9990), range (0.02–2.40 mg L⁻¹; 0.15-24.00 mg L⁻¹), LOD (3.0–4.8 μg L⁻¹; 30.1–48.0 μg L⁻¹), LOQ (9.9–15.8 μg L⁻¹; 150.5-240.0 μg L⁻¹), intra-day precision (4.4-7.1% RSD; 3.3-5.1% RSD), accuracy (94.06-113.7%; 89.5-108.4%), retention time repeatability (0.0-0.5% RSD; 0.0-0.6% RSD), and peak area repeatability (1.2-4.0% RSD; 2.4-3.5% RSD) for positive and negative ion detection modes, respectively. The statistical comparison of the quantitative results obtained in positive and negative ion detection mode was performed.     

Analysis of mycotoxin fumonisins in corn products by high-performance liquid chromatography coupled with evaporative light scattering detection

Fumonisins are mycotoxins produced by the fungus Fusarium verticillioides, which is a widespread pathogen of corn. The mycotoxins are known to cause fatal diseases in some domestic animals and have been linked to human esophageal cancer in China and South Africa. Here, we describe a simple method for direct and quantitative analysis of the toxins in food products. The method is based on high-performance liquid chromatography coupled with an evaporative laser scattering detector (HPLC–ELSD) without any prior derivatization of the samples. Using this method, we have analyzed corn-based food samples from central markets in eastern China. The results showed that FB₁ was the main contaminant in the samples. The overall level of fumonisin contamination was relatively low, with a range of 0.25–1.80 μg/g (mean 0.74 μg/g) in 66.7% (16 of 24) of corn samples from Middle-eastern Area, 0.21–0.29 μg/g (mean 0.24 μg/g) in 28.6% (6 of 21) of corn samples from Northeastern Area, and 0.30–3.13 μg/g (mean 0.47 μg/g) in 30.0% (6 of 20) of corn samples from Southeastern Area.     

Antioxidant activity and phenolic content of 16 raisin grape (Vitis vinifera L.) cultivars and selections

Six raisin grape cultivars and 10 new raisin grape selections were analyzed for antioxidant activity (ABTS assay) and for total and individual phenolic compounds. Samples were freeze–dried and values are reported on a dry weight basis. Antioxidant activity across the 16 samples ranged from 7.7 to 60.9 μmol Trolox/g DW, with A95-27 exhibiting the greatest activity. Total phenolic content, determined in gallic acid equivalents using the Folin–Ciocalteau assay, ranged from 316.3 to 1141.3 mg gallic acid/100 g DW and was strongly correlated (r = 0.990) with antioxidant results. Concentrations of individual phenolics were determined by HPLC. trans-Caftaric acid was the predominant compound in all samples. A95-15 contained the lowest concentration (153.5 μg/g DW) of caftaric acid, while Fiesta contained the highest concentration (598.7 μg/g DW). Selections A56-66, A95-15, and A95-27 had much higher levels of catechin (86.5–209.1 μg/g DW) and epicatechin (126.5–365.7 μg/g DW) than the other samples.     

Effect of refining processes on antioxidant capacity,total contents of phenolics and carotenoids in palm oils

Antioxidant capacity (AC), total phenolic content (TPC) and total carotenoid content (TCC) in palm oils at various stages of the refining process from two technological modes were determined. The obtained mean FRAP and DPPH values for the methanolic extracts of palm oils from mode 1 (19.5–102.8 μmol TE/100 g and 18.8–103.0 μmol TE/100 g) were lower than for oils from mode 2 (25.6–134.8 μmol TE/100 g and 25.4–135.4 μmol TE/100 g). The total phenolics (4.1–12.4 mg GA/100 g) and carotenoids (0.18–45.8 mg/100 g) in the studied oils were correlated with their antioxidant capacities determined by FRAP and DPPH methods (r = 0.6623–0.9878). During the refining process, for both technological modes resulted in a loss of AC by 80%, TPC by 26–55% and TCC by 99%. The bleaching step caused the highest losses of AC as determined by FRAP 41% and 46%, DPPH by 43% and 48%, while TPC loss was 45% and 23% and loss of carotenoids was 49% and 56%, in mode 1 and mode 2, respectively.     

Determination of trace metals in canned fish marketed in Turkey

The levels of trace metals of canned fish samples collected from markets in Turkey were determined by flame and graphite furnace atomic absorption spectrometry after microwave digestion. The accuracy of the method was corrected by standard reference material (NRCC-DORM-2 Dogfish Muscle). The contents of investigated trace metals in canned fish samples were found to be in the range 1.10–2.50 μg/g for copper, 7.57–34.4 μg/g for zinc, 0.90–2.50 μg/g for manganese, 10.2–30.3 μg/g for iron, 0.96–3.64 μg/g for selenium, 0.45–1.50 μg/g for aluminium, 0.97–1.70 μg/g for chromium, 0.42–0.85 μg/g for nickel, 0.09–0.40 μg/g for lead and 0.06–0.25 μg/g for cadmium. The results were compared with the literature values.     

Quantification of coumarin derivatives in Noni (Morinda citrifolia) and their contribution of quenching effect on reactive oxygen species

The quantification of coumarin derivatives such as scopoletin, 7-hydroxycoumarin (7-HC) and 4-hydroxycoumarin (4-HC) in Noni (Morinda citrifolia) was described. The coumarin derivatives were determined by HPLC-UV or -fluorescence detection. More than 95% of peak purity for coumarin derivatives in Noni sample was confirmed by a multi-wavelength fluorescence detector. Amounts of scopoletin and 7-HC in Noni juices (A–H) were ranging 5.1–231 μg/ml and 0.04–0.45 μg/ml, respectively (n = 12). No 4-HC was detected in any Noni samples examined.     

Trace metals in raw cows’ milk and assessment of transfer to Comté cheese

As part of a program to assess the transfer of metals from soil to dairy products, the transfer of metal trace elements to milk and cheese was studied. Concentrations of non-essential (Cd and Pb) and essential elements (Cu and Zn) were determined by atomic absorption spectrometry in 61 samples of raw milk and 21 of the corresponding cheese. While metal concentrations (dry weight) in raw milk were very low (Cd: 0.34–1.01 ng/g; Pb: 0.009–0.126 μg/g; Cu: 0.28–1.71 μg/g; Zn: 20.62–30.96 μg/g), concentrations in the corresponding cheese were significantly higher (Cd: 0.68–11.37 ng/g; Pb: 0.020–0.925 μg/g; Cu: 5.35–21.34 μg/g; Zn: 33.66–63.41 μg/g). The retention factor R_t suggests a concentration effect during the cheese making process, especially in the case of Cu, due to the use of large copper vats. Concentrations of non-essential elements (Cd and Pb) in cheese largely remained below those considered as dangerous for consumers. Finally, Comté cheese may constitute a useful source of Cu and Zn in human diet.     

Essential oil composition,antimicrobial and antioxidant activities of Satureja cuneifolia Ten.

Satureja cuneifolia Ten. is a well-known aromatic plant which is frequently used as a spice and herbal tea in Anatolia. S. cuneifolia oil was analyzed by gas chromatography/mass spectrometry (GC/MS). The major components of S. cuneifolia oil were carvacrol (44.99%) and p-cymene (21.61%). The essential oil of S. cuneifolia exhibited antimicrobial activity against all of the tested foodborne and spoilage bacteria. The minimum inhibitory concentration (MIC) values for test bacteria which were sensitive to the essential oil of S. cuneifolia were in the range of 600–1400 μg/ml. Antioxidant activities of the essential oil and the methanolic extract from S. cuneifolia were evaluated by using DPPH radical scavenging, β-carotene–linoleic acid bleaching and metal chelating activity assays. In addition, the amounts of total phenol components in the plant methanolic extract (222.5 ± 0.5 μg/mg) and the oil (185.5 ± 0.5 μg/mg) were determined.     

Natural occurrence of deoxynivalenol in wheat from Paraná State,Brazil and estimated daily intake by wheat products

The occurrence of deoxynivalenol (DON) was evaluated in 113 wheat samples from the northern and central/southwestern regions of Paraná State, Brazil during the 2008 and 2009 growing seasons, and this rate of occurrence was used to estimate the DON dietary exposure. The DON determination was carried out by an indirect competitive enzyme-linked immunosorbent assay. DON was detected in 66.4% samples at levels ranging from 206.3 to 4732.3 μg/kg (mean 1894.9 μg/kg). The estimated daily intake (EDI) of DON through bread and pasta was evaluated in the inhabitants of Londrina City in northern Paraná State, Brazil. The average intake of these inhabitants was 0.79 μg/kg body weight (b.w.) for bread and 0.35 μg/kg b.w. for pasta. The total EDI was 1.13 μg/kg, which is above the Provisional Tolerable Daily Maximum Intake (PTDMI) of 1 μg/kg b.w. To our knowledge, this is the first report on natural DON occurrence in wheat and DON dietary exposure estimation from Paraná, Brazil.     

Inhibitory effect of marinades with hibiscus extract on formation of heterocyclic aromatic amines and sensory quality of fried beef patties

Heterocyclic aromatic amines (HAA) are carcinogenic compounds found in the crust of fried meat. The objective was to examine the possibility of inhibiting HAA formation in fried beef patties by using marinades with different concentrations of hibiscus extract (Hibiscus sabdariffa) (0.2, 0.4, 0.6, 0.8 g/100 g). After frying, patties were analyzed for 15 different HAA by HPLC-analysis. Four HAA MeIQx (0.3–0.6 ng/g), PhIP (0.02–0.06 ng/g), co-mutagenic norharmane (0.4–0.7 ng/g), and harmane (0.8–1.1 ng/g) were found at low levels. The concentration of MeIQx was reduced by about 50% and 40% by applying marinades containing the highest amount of extract compared to sunflower oil and control marinade, respectively. The antioxidant capacity (TEAC-Assay/Folin–Ciocalteu-Assay) was determined as 0.9, 1.7, 2.6 and 3.5 μmol Trolox antioxidant equivalents and total phenolic compounds were 49, 97, 146 and 195 μg/g marinade. In sensory ranking tests, marinated and fried patties were not significantly different (p > 0.05) to control samples.     

Analysis of multi-pesticide residues in the foods of animal origin by GC–MS coupled with accelerated solvent extraction and gel permeation chromatography cleanup

A new analytical method was developed to simultaneously determine residues of 109 pesticides (including isomers) in the foods of animal origin. Acetonitrile was selected for accelerated solvent extraction (ASE) for effectively extracting the pesticides from the fatty samples. The cleanup was performed with an automated gel permeation chromatography (GPC) cleanup system. The prepared samples were analysed with GC–MS in the selected ion monitoring mode (SIM) using one target and two qualitative ions for each analyte. Chlorpyrifos-d₁₀ was used as an internal standard. The lowest limit of detection was 0.3 μg kg⁻¹ for some pesticides. The recoveries and relative standard deviations (RSDs) were checked by spiking untreated samples with pesticides at 0.05, 0.1 and 0.2 mg kg⁻¹. The average recoveries of most pesticides were from 62.6% to 107.8%. The precision values expressed as RSD were all ?20.5% (n = 6). Good linearity (r ? 0.99) was observed between 0.05 and 1.5 μg mL⁻¹.     

Fatty acids and cholesterol oxidation in salted and dried shrimp

The aim of this work was to evaluate the occurrence of cholesterol oxidation products and to analyze the lipidic profile in salted–dried shrimp. Fifty samples of salted–dried shrimp were evaluated, and the cholesterol oxides (7β-OH, 7α-OH, 7-Keto and 25-OH) were quantified by high-performance liquid chromatography. The cholesterol oxides: 7β-OH (34.63–72.56 μg/g), 7α-OH (5.02–12.12μg/g), 7-Keto (7.44–32.68 μg/g) and 25-OH (2.37–22.88 μg/g) were determined in all samples analyzed. Regarding to the total cholesterol content and the average thiobarbituric acid reactive substances (TBARS) content, the results ranged from 73.88 to 247.69 mg/100 g, and 0.02 to 1.30 mgMA/kg, respectively. The fatty acids profile was: 27.48% saturated, 43.90% monounsaturated and 28.61% polyunsaturated. The presence of cholesterol oxidation products and the values of TBARS indicate the degree of oxidation of this product, which was probably initiated by inadequate conditions of processing and storage.     

Optimization of gas chromatography–single quadrupole mass spectrometry conditions for multiresidue analysis of pesticides in grapes in compliance to EU-MRLs

A single quadrupole GC–MS method was optimized for multiresidue determination of 47 pesticides in grapes with limit of quantifications of each compound in compliance with the EU-MRL requirements. Sample preparation involved extraction of 10 g sample with 10 ml ethyl acetate (+10 g sodium sulphate) by homogenization at 15,000 rpm followed by centrifugation at 3000 rpm. The supernatant was cleaned by dispersive solid phase extraction with primary secondary amine and acidified with 0.1% formic acid. Residues were estimated in selected ion monitoring mode with programmable temperature vaporizer-large volume injection (8 μl). All the GC and MS parameters were thoroughly optimized to achieve satisfactory linearity (R² > 0.99) within 0.01–0.25 mg kg⁻¹ with minimum matrix interferences. Recoveries at 0.01 and 0.02 mg kg⁻¹ were within 67–120% with associated precision RSD below 19%. The method was successfully applied for analysis of the real world samples for incurred residues.     

Detection of alkaline phosphatase by competitive indirect ELISA using immunoglobulin in yolk (IgY) specific against bovine milk alkaline phosphatase

Immunoglobulin in yolk (IgY) (with a titer of 1.3 × 10⁶) specific against bovine milk (BM) alkaline phosphatase (ALP) was obtained by intramuscularly immunizing hens on the thigh and was used as the primary antibody to conduct competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) to determine BMALP in ALPs from BM and Escherichia coli sources. A relationship between the ELISA value and the BMALP level (0.01–10 μg/mL) in whole milk (R² = 0.9019) or in skimmed milk (R² = 0.9402) was observed. The maximal inhibition (%) of BMALP on the microtiter plate by free BMALP at 10 μg/mL whole milk (3.89 mU/μg BMALP) was about 50%, while no inhibition (%) of BMALP by free E. coli ALP at concentrations between 0.01 to 10 μg/mL (60 mU/μg E. coli ALP) was determined. At BMALP levels higher than 0.1 μg/mL, CI-ELISA was proved to be effective in differentiating between BMALP and E. coli ALP and quantifying BMALP in whole milk or skimmed milk in the presence of E. coli ALP with an activity of 0.6 U/mL. Higher inhibition (about 70%) of BMALP on the microtiter plate by free BMALP in diluted (10¹–10⁴ fold) milk samples was observed. The optimal conditions for CI-ELISA in determining BMALP (0.1–10 μg/mL) from ALPs in milk samples were using 10³-fold diluted crude IgY specific against BMALP as primary antibody and 10³-fold diluted goat anti-chicken IgG–ALP conjugate as the secondary antibody.     

Quantitative analysis of acrylamide in tea by liquid chromatography coupled with electrospray ionization tandem mass spectrometry

An effective sample preparation procedure was optimized and a liquid chromatography–tandem mass spectrometry (LC–MS/MS) was developed for the quantitative analysis of acrylamide in tea. [¹³C₃]-acrylamide was used as internal standard. Acrylamide was extracted at 25 °C for 20 min by 10 ml water followed by 10 ml acetonitrile, and then 4 g of magnesium sulfate and 0.5 g of sodium chloride were added to the above mixture under stirring thoroughly. In order to increase the response of acrylamide, 9 ml acetonitrile layer was taken and concentrated to 0.5 ml. Solid-phase extraction with an Oasis MCX cartridge was carried out for clean-up. The limit of detection (LOD) and limit of quantification (LOQ) were 1 and 5 ng/ml, respectively. The recovery efficiency of the extraction procedure ranged between 74% and 79%. The levels of acrylamide in 30 tea samples were less than 100 ng/g. Black, oolong, white and yellow tea samples had quite low acrylamide contents (<20 ng/g). Higher acrylamide levels occurred in baked, roasted, and one sun-dried green tea samples (46–94 ng/g).     

Antimicrobial properties and analytical profile of traditional Eruca sativa seed oil: Comparison with various aerial and root plant extracts

The present study investigates the antimicrobial activity of various solvent extracts of Eruca sativa (aerial and root) and seed oil against-antibiotic resistant Gram-negative (Escherichia coli, Pseudomoms aeruginosa and Shigella flexneri) and Gram-positive (Staphylococcus aureus and Bacillus subtilis) bacteria. Among the various preparations, seed oil was the most active, exhibiting a maximum zone inhibition of 97% for Gram-positive bacteria and of 74–97% for Gram-negative bacteria. The MIC of the seed oil was found to be 65–75 and 60–70 μg/ml for Gram-negative and Gram-positive bacteria, respectively. Analytical investigation on main volatile and non-volatile components was performed on seed oil. Among the formers allyl isothiocyanate (40 μg/g), 3-butenyl isothiocyanate (260 μg/g), 4-methylsulfinybutyl isothiocyanate (sulforaphane 743 μg/g), 2-phenylethyl isothiocyanate (159 μg/g) and bis(isothiocyanatobutyl)disulphide (∼5000 μg/g) were determined by head space/SPME/GC–MS analysis. Free fatty acids were 1.6% w/w of the oil and overall 25 fatty acids were identified. Erucic and oleic acids were the main fatty acids both in the free (7.8 and 2.1 mg/ml) and esterified forms (50.6% w/w and 14.9% w/w of total fatty acids). Unsaponifiable fraction was 1.8% w/w.