Flavonoid,tannin and hypericin concentrations in the leaves of St. John’s wort (Hypericum perforatum L.) are affected by UV-B radiation levels

St. John’s wort (Hypericum perforatum L.) herb is used as a herb-tea or as a food supplement because of its antidepressive properties. St. John’s wort, grown under different levels of UV-B radiation, was analysed for its concentrations of flavonoids, tannins and hypericin. A high level of UV-B radiation increased leaf concentrations of flavonoids from 6.31 to 9.00/100 g in dry matter (DM) and of tannins from 26.6 to 31.4/100 g in DM. The concentration of hypericin in leaves exposed to enhanced UV-B radiation (0.08/100 g in DM) was lower than that of leaves exposed to reduced UV-B radiation (0.09/100 g in DM). We conclude that the concentrations of bioactive substances in St. John’s wort herb can be altered by regulating the environment during plant growth.     

Selenium species in buckwheat cultivated with foliar addition of Se(VI) and various levels of UV-B radiation

Common (Fagopyrum esculentum Moench) and tartary (Fagopyrum tataricum Gaertn.) buckwheat was treated by spraying the leaves with a water solution containing 15 mg Se per litre in the form of sodium selenate in the flowering period. The selenium content in all parts of plant was found to be less than 200 ng g⁻¹ in non-treated and in the range 2700–4650 ng g⁻¹ in selenium treated buckwheat. Exposure to UV-B radiation lead to higher Se accumulation in flowers of both Se enriched cultivars. For speciation analysis enzymatic hydrolysis was carried out, separation and detection of selenium species was performed by high performance liquid chromatography–ultraviolet treatment–hydride generation atomic fluorescence spectrometry (HPLC–UV–HG-AFS). In flowers and leaves, on average 11% of the Se content was soluble and in the form of Se(VI), representing between 0.6% (flowers) and 3% (leaves) of the Se content. The remaining soluble non-amino acid organic Se was not detected by HPLC–UV–HG-AFS. In seeds 93% of the selenium content was found in the extracts and the main selenium species was SeMet with 93 ± 5% relative to the selenium content.     

Determination of ionophores in raw bovine milk using LC–MS/MS: Application to residue surveillance

Residues of four ionophores (lasalocid, monensin, narasin, and salinomycin) in raw milk samples were extracted with acetonitrile and subsequently determined using liquid chromatography–tandem mass spectrometry. Ionophores could be determined down to 0.1 ng g⁻¹ level, without additional cleanup or concentration of the resulting extract. The analysis of a series of raw milk samples fortified at analyte concentrations ranging from 1 to 20 ng g⁻¹ yielded average accuracies ranging from 60.7% to 118.3% with percent relative standard deviations below 13%. During six months of a surveillance program, 1072 raw milk samples were collected from the transport chain of dairy producers in Alberta and analysed. Monensin was detected in 736 of 1072 samples tested at concentrations ranging from 0.10 to 0.53 ng g⁻¹ which is well below the current Canadian maximum residue limit of 10 ng g⁻¹.     

Dispersive liquid–liquid microextraction combined with sweeping micellar electrokinetic chromatography for the determination of some neonicotinoid insecticides in cucumber samples

A rapid, simple and sensitive method has been developed for the analysis of some neonicotinoid insecticides in cucumber samples by using dispersive liquid–liquid microextraction (DLLME) coupled with sweeping in micellar electrokinetic chromatography (MEKC). Under optimised conditions, the enrichment factors were achieved in the range from 4000 to 10,000. The linearity of the method was in the range from 2.7 to 200 ng g⁻¹ for thiacloprid, acetamiprid and imidacloprid, and in the range from 4.0 to 200 ng g⁻¹ for imidaclothiz in cucumber samples, with the determination coefficients (r²) ranging from 0.9924 to 0.9968. The limits of detection (LODs, S/N = 3) ranged from 0.8 to 1.2 ng g⁻¹. The relative standard deviations (RSDs) at the concentration levels of 10.0 and 50.0 ng g⁻¹ each of the neonicotinoid insecticides in cucumber samples varied from 3.8% to 6.3%. The developed method has been successfully applied to the analysis of the neonicotinoid insecticides in cucumbers with a satisfactory result.     

Quantification of cadmium and lead in offal by SF-ICP-MS: Method development and uncertainty estimate

Offal of bovine, ovine and porcine are able to accumulate potentially toxic heavy metals, such as Cd and Pb, posing a risk for human health. For this reason, the Commission Regulation no. 466/2001 provided the maximum admitted levels for these metals in this kind of matrix (Cd, 500 ng g⁻¹ in all kinds of offal; Pb, 500 ng g⁻¹ in liver and 1000 ng g⁻¹ in kidney). A method based on sector field inductively coupled plasma mass spectrometry for Cd and Pb quantification in calf liver was developed and further applied to offal of different animals. The uncertainty of measurements was calculated according to the Eurachem/Citac Guide. The method LoDs and LoQs were 3.5 and 11 ng g⁻¹ for Cd, and 2.0 and 6.0 ng g⁻¹ for Pb. The repeatability and the intra-laboratory reproducibility showed relative standard deviations equal to 2.25% and 1.99% for Cd and 1.49% and 6.55% for Pb. Relative expanded uncertainties at the mean value in calf liver were 4.74% for Cd and 13.8% for Pb. In Italian offal the following concentration intervals were found (in ng g⁻¹): (i) Cd: calf, from <3.5 in spleen and lung to 96.4 in kidney; lamb, <3.5 in all offal; pig, 114 in liver and (ii) Pb: calf, from 4.03 in lung to 31.8 in liver; lamb, from 4.71 in heart to 279 in liver; pig, 9.19 in liver.     

Total content of As,Sb, Se,Te and Bi in Spanish vegetables,cereals and pulses and estimation of the contribution of these foods to the Mediterranean daily intake of trace elements

As, Sb, Se, Te and Bi were quantified in vegetables, pulses and cereals, in order to increase the available information on the presence of these elements in the Mediterranean daily intake. Samples were dry ashed and the ashes dissolved with diluted HCl. Hydride generation atomic fluorescence spectrometry was used for quantification. Limits of detection (LOD) ranged from 0.2 to 1.7 ng g⁻¹ and the accuracy was assessed by the analysis of three certified reference materials, Rice Flour (NIST 1568a), Tomato Leaves (NIST 1573) and Cabbage (IAEA 359). Good concordance between results found and certified values were always observed. As, Sb, Se, Te and Bi levels varied in samples analysed from values under the LOD till 1001, 47, 270, 104 and 48 ng g⁻¹, respectively.     

Mutagenic heterocyclic amine content in thermally processed offal products

Heterocyclic amines (HAs) are potent mutagens formed during heat-processing of proteinaceous food. PhIP is the most ubiquitous and abundant mutagenic HA. In this study several offal products (beef liver, lamb kidney and beef tongue) have been thermally processed and analysed for HAs for the first time. Norharman and harman were the amines most abundant, found at concentrations below 2 ng g⁻¹. PhIP was only formed up to 0.12 ng g⁻¹. Among the rest of HAs analysed, only DMIP, MeIQx and 4,8-DiMeIQx were detectable in cooked kidney and tongue up to 0.25 ng g⁻¹. The influence of cooking additives on HAs formation was evaluated, finding higher levels of norharman and harman, up to 8.87 ng g⁻¹, in liver processed with additives, whereas similar levels of the rest of HAs were found in these samples. The low amounts of HAs found after the prolonged thermal treatments over 200 °C indicates that offal products are among the types of meat that produce lowest exposure to HA.     

Immunoassay based on monoclonal antibody for aflatoxin detection in poultry feed

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on an anti-aflatoxin B₁ monoclonal antibody was standardised and validated for aflatoxin screening in poultry feed samples and its performance was compared to high-performance liquid chromatography (HPLC). The ic-ELISA showed good linearity (r² = 0.994) and detection limits of 1.25 ng g⁻¹ for broiler feed and 1.41 ng g⁻¹ for laying hen feed. Mean aflatoxin recovery rates by ic-ELISA were 102% (laying hen feed) and 98% (broiler feed). Aflatoxins were detected in 88.2% of the 34 broiler feed samples by ic-ELISA and HPLC at means of 10.48 ng g⁻¹ and 8.41 ng g⁻¹, respectively, while 92% of laying hen feed samples (n = 36) showed aflatoxin contamination at means of 20.83 and 19.75 ng g⁻¹. The standardised ic-ELISA showed reliability and a high correlation with HPLC of 0.97 (broiler feed) and 0.98 (laying hen feed) indicating its potential for aflatoxin screening in poultry feed samples.     

Use of a portable time-resolved fluorometer to determine oxytetracycline residue in four fruit crops

Oxytetracycline (OTC) is used worldwide to protect crops against bacterial diseases. The US Environmental Protection Agency approved its use in apple, pear, nectarine, and peach, and set residue tolerance at 350 ng g⁻¹. A europium-sensitised luminescence (ESL) method was developed for in-situ determination of OTC residue in these fruits. After extraction in Na₂EDTA-NaCl-McIlvaine buffer at pH 4 and filtration, cleanup was performed using hydrophilic-lipophilic balance cartridges. ESL was measured using a portable time-resolved fluorometer. The signal responded linearly over three orders of magnitude (10–10000 ng g⁻¹) with 17–50 ng g⁻¹ limits of quantitation and 2% averaged relative standard deviation. Recoveries were 84% and 82% at 100 and 350 ng g⁻¹, respectively. Inter-laboratory validation was performed by HPLC–MS/MS.     

Effects of ageing and low internal temperature of grilling on the formation of heterocyclic amines in beef Longissimus dorsi muscle

The mutagenic heterocyclic amines (HAs) originate in processed proteinaceous food. The effects of ageing (non-aged - i.e. 24 h post mortem vs. 14 and 28 days post mortem kept at 1 °C) and final internal temperature on cooking (T_i, 65 and 80 °C) on the content of HAs in grilled steaks (two-plated grill, temperature of 220 °C) were studied. HA precursors (creatine, creatinine, and free amino acids) and ageing indicators, such as instrumentally measured colour values, pH_ultimate values and length of myofibrilar fragments on raw and cutting values on grilled beef Longissimus dorsi muscles were determined. The muscles originated from eight commercially slaughtered Simmental bulls, 19-20 months old. The content of HAs was determined by a solid-phase extraction procedure. Meat ageing is accompanied by large changes in the chemical composition and structure of muscle tissues. In general, all the ageing indicators and precursors of HAs were influenced by ageing time at the 5% level or less. Creatine content declined significantly (non-aged: 6.00 mg g⁻¹, 14 days: 5.82 mg g⁻¹, and 28 days: 5.55 mg g⁻¹) and creatinine increased with days of ageing (non-aged: 0.19 mg g⁻¹, 14 days: 0.24 mg g⁻¹, and 28 days: 0.26 mg g⁻¹). Higher contents of total free amino acids were determined after 14 and 28 days of storage (28.18 μmol g⁻¹ and 37.59 μmol g⁻¹) than in non-aged beef (19.00 μmol g⁻¹). In this study, two HAs were determined: MeIQx (2-amino-3,8-dimethyl-imidazo[4,5-f]quinoxaline) and PhIP (2—amino-1-methyl-6-phenylimidazo-[4,5-b]pyridine). The content of HAs increases with ageing. At lower T_i, more MeIQx was formed; at higher T_i, more PhIP was formed. MeIQx was present in all samples while PhIP was found only in samples grilled to higher T_i. Samples treated to T_i = 80 °C generally contained less HAs (non-aged meat: 0.20 ng g⁻¹, 14 days: 0.26 ng g⁻¹, and 28 days: 0.28 ng g⁻¹) than samples treated to T_i = 65 °C (non-aged meat: 0.19 ng g⁻¹, 14 days: 0.36 ng g⁻¹, and 28 days: 0.39 ng g⁻¹) on account of MeIQx thermolability.     

Determination of aflatoxins in eggs,milk, meat and meat products using HPLC fluorescent and UV detectors

Raw and pasteurised sheep’s, cow’s and goat’s milk, eggs, and beef samples from different local markets in Jordan were collected during a period of 5 months (January through May 2007) and examined for aflatoxins B1(AFB1), B2(AFB2), G1(AFG1), G2(AFG2), M1(AFM1) and M2(AFM2). The samples were analysed with high performance liquid chromatography (HPLC) using UV and Fluorescent detectors. The analysed samples of milk collected in January were found to contain 0.56 μg L⁻¹ AFM1 and 0.1 μg L⁻¹ AFM2 whilst, the concentration of AFM1 and AFM2 was < 0.05 μg L⁻¹ for milk samples collected between March and May. The AFB1, AFB2, AFG1 and AFG2 contents in the analysed food products ranged from 1.10 to 8.32 μg L⁻¹ and 0.15 to 6.36 μg L⁻¹ in imported and fresh meat samples collected during March, respectively. The mean recovery for the HPLC method was 92% to 109% and the quantification levels were 50 ng L⁻¹ for AFM1 and AFM2. The AFM1 was found in 10% of the tested samples with concentrations between 0.08 and 1.1 μg kg⁻¹ and AFM2 was only found in 1.82% of the tested samples with a level of 0.1 μg kg⁻¹. The AFM1 levels in the examined foods were higher than the maximum level of AFM1 in liquid milk set by the European Community and Codex Alimentarius of 50 ng L⁻¹.     

Heterocyclic amines and azaarenes in pan-fried meat and its gravy fried without additives and in the presence of onion and garlic

The effect of onion and garlic on the formation of heterocyclic amines (HAs, aminoazaarenes) and azaarenes (aza-PAHs) was evaluated by comparing the concentrations of several compounds in meat and gravy samples, obtained from three pork dishes prepared in the presence and absence of these spices. The concentrations of individual HAs (8-MeIQx, MeIQ, 4,8-DiMeIQx, PhIP) were from 0.5 ng g⁻¹ to 10.5 ng g⁻¹ of meat and of azaarenes (benzo[a]acridine, benzo[c]acridine, dibenzo[a,c]acridine, dibenzo[a,j]acridine and dibenzo[a,h]acridine) – from 0.06 ng g⁻¹ to 0.99 ng g⁻¹. The addition of onion (30 g/100 g of meat) in the dishes investigated, caused a decrease in heterocyclic amines concentration (considering total contents in meat and gravy) in the range of 31–49% and of azaarenes by 21–48%. Garlic (15 g/100 g of meat) lowered the concentration of HAs by 26–36% and azaarenes by 33–40%; the changes in concentrations caused by these spices were different for particular compounds. Components of onion and garlic intensify the extraction of heterocyclic amines and azaarenes from meat in gravy.     

Arsenic removal from rice by washing and cooking with water

Two Hungarian and one Chinese rice samples were selected in order to establish the extractable arsenic content by washing and cooking in water in a ratio of 6:1, water:rice (cm³:g) by inductively coupled plasma sector field mass spectrometry (ICP-SF-MS). Total arsenic concentration of the Zhenshan 97, Risabell and Ko?röstáj-333 samples were 171.3 ± 7.1 ng g⁻¹, 116.0 ± 3.7 ng g⁻¹ and 139.0 ± 6.1 ng g⁻¹, respectively, which did not exceed the toxic limits established for As in Hungary (0.3 μg g⁻¹) or in China (0.7 μg g⁻¹). The predominant chemical form of As in the raw rice samples determined by on-line high performance liquid chromatography and ICP-MS was arsenite. Moreover, enzymatic hydrolysis with α-amylase + protease and microprobe focused sonication proved that arsenite could be removed in the highest extent by washing and cooking, meanwhile the main As form remaining in the cooked rice was As(V). Thus, it is recommended to prepare rice-containing dishes in abundant water, which should be discarded after washing and cooking. The results were validated with a NIST SRM 1568a.     

Oral bioaccessibility and human risk assessment of organochlorine pesticides (OCPs) via fish consumption,using an invitro gastrointestinal model

The oral bioaccessibility of dichlorodiphenyltrichloroethane (DDT) and hexachlorocyclohexanes (HCHs) in twenty fish species were evaluated using an in vitro gastrointestinal model. Relatively high concentrations of ∑HCH (0.33–9.88 ng g⁻¹, mean 1.57 ng g⁻¹) and ∑DDT (0.74–131 ng g⁻¹, mean 12.2 ng g⁻¹) were observed in all market fish. The average bioaccessibilities for ∑HCHs were 3.35% and 8.73% in gastric and intestinal conditions, respectively, and for ∑DDTs were 5.48% and 17.6%, respectively. Significant (p < 0.05) correlations were observed between OCP congener digestible concentrations in fish muscle and their corresponding concentrations in human tissues of Hong Kong residents. The health risk assessments based on solvent and digestible concentrations were found to be contradictory due to the fact that only a limited proportion of OCP was bioaccessible. It was concluded that human health risk assessment based on solvent concentrations should be modified by taking bioaccessibility of the contaminant into account.     

Highly selective optical nitrite sensor for food analysis based on Lauth’s violet-triacetyl cellulose membrane film

A new sensor has been developed for the determination of nitrite based on the immobilisation of Lauth’s violet on triacetyl cellulose membrane using absorption spectrophotometry. The optical sensor signal is based on the reaction of Lauth’s violet with bromate as an oxidant in the presence of nitrite to produce a colourless product. Nitrite has a strong catalytic effect on the oxidation of Lauth’s violet with bromate in acidic media. The difference in the absorbance of the optode at 617 nm between uncatalysed and catalysed reactions (ΔA) was directly proportional to the concentration of nitrite in sample solution. A linear calibration curve (r² = 0.985) was observed in the nitrite concentration range of 10.12-1012 ng mL⁻¹ and a detection limit of 8.3 ng mL⁻¹ was found.     

Application of acetylacetone chelation solid-phase extraction to GFAAS measurements of trace amounts of beryllium in livers and muscles of poultry and livestock

Trace amounts (0.004–0.55 ng) of beryllium (Be) in a dried bovine liver sample (20 mg) can be accurately determined by graphite-furnace atomic absorption spectrophotometry (GFAAS) after treating with microwave digestion (HNO₃/H₂O₂) at 85 °C for 10 min and using acetylacetone as a chelating agent in the presence of an acetate buffer (pH 6.0). The method detection limit (MDL, 3σ) for Be was found to be 0.18 ng g⁻¹ and the limit of quantification (LOQ, 10σ) was found to be 0.60 ng g⁻¹; the calibration graph was linear up to 27 ng g⁻¹. The Be contents measured in four liver and four muscle samples (BCR CRM-185R bovine liver, BCR CRM-384 pork muscle, and six samples collected in Kaohsiung, Taiwan, ROC) were between 2.3 and 4.7 ng g⁻¹. Good spiked recoveries (96.0–103.0%) were obtained for these eight samples with a relative standard deviation (RSD, n = 3) ?3.0%. The method could be applied to measurements of Be in livers and muscles of poultry and livestock.     

Enhanced selenium content in buckwheat (<Emphasis Type="Italic">Fagopyrum esculentum</Emphasis> Moench) and pumpkin (<Emphasis Type="Italic">Cucurbita pepo</Emphasis> L.) seeds by foliar fertilisation

The fruit and thin-husked seeds of the pumpkin (Cucurbita pepo L.) and buckwheat grain (Fagopyrum esculentum Moench), both grown in Slovenia, were analysed for selenium (Se) content following foliar application of Se(VI) solution during the period of blooming. Samples were digested by a H₂SO₄-HNO₃-H₂O₂-V₂O₅ mixture and Se determined, based on hydride generation atomic fluorescence spectrometry. The whole procedure from weighing to measuring was carried out in the same Teflon vessel. The detection limit of the method was 0.14 ng g^–1 solution. Buckwheat seeds from untreated plants contained 47 ng g^–1 of Se and 394 ng g^–1 from plants after foliar fertilisation with Se. Pumpkin seeds from untreated plants contained 108 ng g^–1 of Se, and 381 ng g^–1 of Se from Se-treated plants, all per lyophilised sample. Se content in lyophilised pumpkin fruit was 15 ng g^–1 in untreated plants and 20 ng g^–1 in Se-treated pumpkin plants. It is thus feasible to enhance Se content in buckwheat and pumpkin seeds by foliar fertilisation, making them a rich source of dietary Se and useful as a raw material for enriched food products.     

Determination of lead and cadmium in food samples by the coprecipitation method

In this study, the coprecipitation method developed using a combination of 2-mercaptobenzothiazole (MBT) as a chelating reagent and copper as coprecipitate carrier was used for the determination of trace lead and cadmium in various food samples by graphite furnace atomic absorption spectrometry (GFAAS). The method was applied for the determination of Pb(II) and Cd(II) in salami, sausage, chicken, anchovy, spinach, cabbage, onion, dill, parsley, lettuce, tea and rice samples. The matrix modifiers were added as 50 μg NH₄H₂PO₄ + 3 μg Mg(NO₃)₂ for both Pb(II) and Cd(II). The signals were measured as peak area. The concentrations of Pb(II) and Cd(II) in the food samples were found to be in the range of 6.63 ng g⁻¹ (anchovy) −3.30 μg g⁻¹ (spinach) and 2.67 ng g⁻¹ (salami) −0.51 μg g⁻¹ (lettuce), respectively.     

Multiresidue determination of tetracycline antibiotics in propolis by using HPLC-UV detection with ultrasonic-assisted extraction and two-step solid phase extraction

An analytical method in propolis was developed and validated for the determination of four tetracyclines (TCs) by high performance liquid chromatography (HPLC) for the first time. After extraction by ultrasound, the extracting solution was subjected to Oasis HLB and weak cation-exchange cartridge to remove water-soluble and fat-soluble flavonoids, aromatic acids, terpenoid compounds, wax, and pollen debris. The calibration curves of fortified samples showed acceptable linear response (R² > 0.99) through a range of 100–5000 ng g⁻¹ in 20 replicates of six concentrations and the analysis of variance (ANOVA) was performed to validate the regression data. The limit of quantification of four TCs were 100 and 150 ng g⁻¹, respectively. The recoveries of the four TCs for propolis samples spiked with 100–500 ng g⁻¹ were in the range of 61.9–88.5% and the RSDs were between 4.80% and 13.2%. Traces of tetracycline were found in two out of 30 analysed real samples.     

Effect of foliar application of selenium on its uptake and speciation in carrot

Carrot (Daucus carota) shoots were enriched by selenium using foliar application. Solutions of sodium selenite or sodium selenate at 10 and 100 μg Se ml⁻¹, were sprayed on the carrot leaves and the selenium content and uptake rate of selenium were estimated by ICP–MS analysis. Anion and cation exchange HPLC were tailored to and applied for the separation of selenium species in proteolytic extracts of the biological tissues using detection by ICP–MS or ESI–MS/MS. Foliar application of solutions of selenite or selenate at 100 μg Se ml⁻¹ resulted in a selenium concentration of up to 2 μg Se g⁻¹ (dry mass) in the carrot root whereas the selenium concentration in the controls was below the limit of detection at 0.045 μg Se g⁻¹ (dry mass). Selenate-enriched carrot leaves accumulated as much as 80 μg Se g⁻¹ (dry mass), while the selenite-enriched leaves contained approximately 50 μg Se g⁻¹ (dry mass). The speciation analyses showed that inorganic selenium was present in both roots and leaves. The predominant metabolised organic forms of selenium in the roots were selenomethionine and γ-glutamyl-selenomethyl-selenocysteine, regardless of which of the inorganic species were used for foliar application. Only selenomethionine was detected in the carrot leaves. The identity of selenomethionine contained in carrot roots and leaves was successfully confirmed by HPLC–ESI–MS/MS.