Analysis of eleven phenolic compounds including novel p-coumaroyl derivatives in lettuce (Lactuca sativa L.) by ultra-high-performance liquid chromatography with photodiode array and mass spectrometry detection.

INTRODUCTION: Lettuce is a widely consumed vegetable and a good source of phenolic compounds. Several factors (genetic, agronomical and environmental) can influence the lettuce composition; their effects are not completely defined and more studies are needed on this topic. OBJECTIVE: To develop an improved ultra-high-performance liquid chromatography (UHPLC) method to quantify the main target intact phenolic compounds in lettuce. METHODOLOGY: UHPLC identification of the compounds was supported by PAD spectra and MS(n) analyses. Quantification was carried out by PAD, by creating matrix-matched calibration curves at the specific wavelength for each compound. RESULTS: Sample pretreatment was simplified, with neither purification nor hydrolysis steps. Chromatographic conditions were chosen to minimise matrix interferences and to give a suitable separation of the major phenolic compounds within 27 min. The method allowed the quantification of 11 intact phenolic compounds in Romaine lettuces, including phenolic acids (caffeoyl and p-coumaroyl esters) and flavonoids (quercetin glycosides). Four p-coumaroyl esters were tentatively identified and quantified for the first time in lettuce. CONCLUSION: The main intact phenolic compounds, including four novel p-coumaroyl esters, were simultaneously quantified in lettuce with optimal performances and a reduced total time of analysis. These findings make headway in the understanding of the lettuce phytochemicals with potential nutritional relevance.

Traceability of sulfonamide antibiotic treatment by immunochemical analysis of farm animal hair samples.

The use of hair to trace use of unauthorized substances, therapeutic agents, or their misuse is becoming very attractive since residues can be detected for a long time after treatment. For this purpose, an indirect enzyme-linked immunosorbent assay (ELISA) has been evaluated for its capability to trace sulfonamide antibiotic treatment by analyzing cattle and pig hair samples. Pigmented and nonpigmented hair samples from control and sulfamethazine (SMZ)-treated pigs and calves were collected, extracted under different alkaline conditions, and analyzed by ELISA after just diluting the extracts with the assay buffer. Data analysis following the European recommendations for screening methods demonstrates that the ELISA can detect SMZ in hair samples with a limit of detection (90% of the zero dose (IC(90))) between 30 and 75 ng g(-1). The same samples have been analyzed by HPLC after a dual solid-phase extraction. The ELISA results matched very well those obtained by the chromatographic method, demonstrating that the immunochemical method can be used as a screening tool to trace animal treatments. Between the benefits of this method are the possibility to directly analyze hair extracts with sufficient detectability and its high-throughput capability. Preliminary validation data are reported using an experimental approach inspired on the Commission Decision 2002/657/EC criteria for screening methods.

Immunochemical assays for direct sulfonamide antibiotic detection in milk and hair samples using antibody derivatized magnetic nanoparticles.

Two direct enzyme-linked immunosorbent assays (ELISAs) have been developed for detection of sulfonamide antibiotic residues in milk samples. One of them is using magnetic nanoparticles (MNP) for target capture/enrichment (Ab-MNP-ELISA), and the second is performed using microtiter plates. Selective polyclonal antibodies, raised against 5-[6-(4-amino-benzenesulfonylamino)-pyridin-3-yl]-2-methyl-pentanoic acid (SA1), used in combination with an enzyme tracer prepared with the same hapten, has allowed us to reach a limit of detection (LOD) lower than 0.5 microg L(-1) for both ELISA formats. Sulfapyridine, sulfamethoxypyridazine, sulfathiazole, and sulfachloropyridazine are detected below the maximum residue limits established by the European Union for these antibiotics in milk (100 microg L(-1)). Matrix effects and accuracy studies performed with full-cream milk and hair extracts indicated a lack of interference from these sample matrices and very good recovery values, especially when using the Ab-MNP format. Milk samples and hair extracts can be measured without any previous treatment. The results demonstrate the high potential of these methods as screening tools for food safety and inspection controls.

Formation of tetracycline degradation products in chicken and pig meat under different thermal processing conditions.

Tetracycline (TC) and 4-epitetracycline (4eTC) degradation, as well as anhydrotetracycline (ATC) and 4-epianhydrotetracycline (4eATC) formation, has been evaluated in thermally treated chicken breast, pig loin, and pig loin with added back-fat. Samples containing TC and 4eTC residues were submitted to microwave or boiling heating, extracted with a mixture of McIlvaine buffer/methanol (75:25), and analyzed by high-performance liquid chromatography-diode array detection on a phenyl-hexyl reverse phase chromatographic column. The formation of ATC and 4eATC, as well as of two unidentified compounds, was described for the first time in edible meat samples submitted to mild thermal treatments, similar to those applied at home to cook foods. Degradation of TC and 4eTC and formation of ATC and 4eATC versus time of treatment fitted satisfactorily a first-order kinetic. Even if the potential toxic effects of these breakdown compounds should be further investigated, their formation in cooked meat should be taken into account when maximum residue limits are established.

Effect of nitrate and nitrite on Listeria and selected spoilage bacteria inoculated in dry-cured ham.

The effect of nitrate and the combination of nitrate/nitrite on Listeria innocua (as surrogate of Listeria monocytogenes). And two selected spoilage microorganisms (Proteus vulgaris and Serratia liquefaciens) was studied in dry-cured ham. Hams were manufactured with different concentrations of curing agents: KNO3 (600 and 150mg/kg) alone or in combination with NaNO2 (600 and 150mg/kg). The addition of 500mg/kg of sodium ascorbate was also evaluated in a batch with 600mg/kg of nitrate and nitrite. The target microorganisms were inoculated by injection in semimembranosus, biceps femoris and in the shank, prior to curing. P. vulgaris and S. liquefaciens were controlled by temperature and aw, respectively, and no effect of nitrate/nitrite was observed. The presence of nitrite in the curing mix reduced L. innocua in semimembranosus, which population was 1.5logcfu/g lower at the end of resting (p<0.05), while at the end of the process it was more frequently detected in the no- and low-nitrite added hams. None of the treatments was able to control Listeria in deeper areas of ham. The addition of sodium ascorbate to the curing mix containing the highest amount of nitrate and nitrite did not show any effect on the microorganisms studied.

Investigation of the aroma of commercial peach (Prunus persica L. Batsch) types by Proton Transfer Reaction-Mass Spectrometry (PTR-MS) and sensory analysis.

The aim of this study was to investigate the aroma and sensory profiles of various types of peaches (Prunus persica L. Batsch.). Forty-three commercial cultivars comprising peaches, flat peaches, nectarines, and canning peaches (pavías) were grown over two consecutive harvest years. Fruits were assessed for chemical aroma and sensory profiles. Chemical aroma profile was obtained by proton transfer reaction-mass spectrometry (PTR-MS) and spectral masses were tentatively identified with PTR-Time of Flight-MS (PTR-Tof-MS). Sensory analysis was performed at commercial maturity considering seven aroma/flavor attributes. The four types of peaches showed both distinct chemical aroma and sensory profiles. Flat peaches and canning peaches showed most distinct patterns according to discriminant analysis. The sensory data were related to the volatile compounds by partial least square regression. γ-Hexalactone, γ-octalactone, hotrienol, acetic acid and ethyl acetate correlated positively, and benzeneacetaldehyde, trimethylbenzene and acetaldehyde negatively to the intensities of aroma and ripe fruit sensory scores.

Immunochemical determination of fluoroquinolone antibiotics in cattle hair: a strategy to ensure food safety.

Enrofloxacin (ERFX) is a synthetic antibiotic of the fluoroquinolone (FQ) family, which is commonly administered in veterinary medicine. ERFX and its metabolite, ciprofloxacin (CPFX), have been reported to accumulate in hair of treated animals. Therefore, hair analysis is an attractive non-invasive alternative to control misuse of such antibiotic and to ensure food safety by preventing such food derived products arrive to the consumer. In this context, an immunochemical analytical protocol has been established to detect ERFX and CPFX residues in cattle hair samples. Unpigmented and pigmented hair were collected from ERFX-treated and non-treated calves, and the aqueous NH4OH extracts were directly analyzed by ELISA, being possible to achieve limits of detection in the range of 10-30 µg kg(-1). A good concordance between HPLC and ELISA measurements was observed. The results demonstrate the potential of the immunochemical procedure reported here to rapidly screen and quantitate FQ residues in hair samples.

Assessment of enrofloxacin and ciprofloxacin accumulation in pig and calf hair by HPLC and fluorimetric detection.

Enrofloxacin is a synthetic bacteriostatic administered in veterinary therapy. It can also be used illegally as a growth promoter to enhance feed efficiency and weight gain. This practice is banned in several countries due to its potential negative effects on the environment and human health. A suitable method for extracting and quantifying enrofloxacin (ENR) and its main metabolite ciprofloxacin (CPR) in cattle and pig hair by high-performance liquid chromatography-fluorimetric detection (HPLC-FLD) had been proposed. ENR and CPR were extracted from hair samples with methanol acidified with trifluoroacetic acid for 24 h at 70 degrees C. The extracts were evaporated and redissolved in the mobile phase before injection. This simplified procedure enabled the detection of both CPR and ENR at ng g-1 levels (limit of detection 4-5 ng g-1) without further purification. Detectable residues of ENR were found in calf and pig hairs after the pharmacological treatment was started. Mean concentrations of quinolone (ENR, CPR) in contaminated hairs ranged from 20 to 2,518 ng g-1 in calves and from 152 to 1,140 ng g-1 in pigs. Hair pigmentation enhanced quinolone accumulation significantly. Hair analysis seems to increase the time window available for the retrospective detection of illegal ENR administration compared to edible tissue analysis.