Use of Multivariate Optimization to Develop Methods for Direct Copper and Lead Determination in Breast Milk by Graphite Furnace Atomic Absorption Spectrometry

A direct method for lead and copper determination in breast milk by graphite furnace atomic absorption spectrometry, using aqueous calibration, was proposed in this study. Samples were diluted with hydroximethylaminomethane 80 %?v/v, pH 8. The dilution determination for Pb and Cu was 1:1 and 1:9, respectively. Fractional factorial (2^4?1) and central composite designs were used to optimize experimental conditions (pyrolysis and atomization temperatures, pyrolysis time, and modifier) using 10 μL samples introduced into a graphite furnace. The methods allowed for copper and lead determination under optimized conditions with an aqueous calibration curve between 0 and 180 μg L^?1 for Cu and 0 and 48 μg L^?1 for Pb. The detection limits were 0.92 μg L^?1 and 6.4 μg L^?1 for Pb and Cu, respectively. Intra and inter-assay studies revealed coefficients of variation of 5.0 and 6.3 %, and 6.4 and 5.5 % for Pb and Cu, respectively. Recovery studies at three concentration levels (three consecutive days, n?=?7/day) presented results between 107 and 109 % for Pb and 102 and 103 % for Cu. Good accuracy was obtained for both metals through recoveries studies using certified reference material (infant formula NIST® 1846). The method determined lead and copper in six samples and the concentrations ranged from 2.90 to 27.9 μg L^?1 for Pb and 384 to 1,212 μg L^?1 for Cu. While copper is an essential element, lead has no nutritional function and is cumulative at low concentrations. Therefore, safe, efficient, and validated methods should be available to determine its concentration in breast milk.     

Cysteamine-CdTe Quantum Dots Electrochemically Synthesized as Fluorescence Probe for Resveratrol

This work proposes a fluorescent probe based on CdTe quantum dots (QDs) for the determination of resveratrol in wine samples. The synthesis of cysteamine (CA) capped CdTe QD was carried out in a one-pot eco-friendly process, resorting to the electrochemical reduction of metallic tellurium powder in a graphite macroelectrode (cavity cell). The reduced species of tellurium (Te^2? and Te₂^2?) migrated to an intermediate compartment of the electrochemical cell and in the presence of a cadmium salt and organic stabilizing agent (CA), forming the colloidal dispersion of CdTe in a single step. Under optimum synthesis conditions, the fluorescence intensity of the prepared nanoparticles varied linearly with the resveratrol concentration in the range from 3.25 to 75 μg L^?1 (R²?=?0.9984), with a detection limit of 0.97 μg L^?1 and RSD of 3.7% (5.0 μg L^?1 resveratrol, n?=?10). The method was successfully applied to the resveratrol determination in wines, with recoveries from 97.8 to 112.4%. Student’s t test was applied and no statistically significant difference was observed between the two methods (HPLC and proposed), with a confidence level of 95% (t_tabulated?=?2.45 and t_calculated?=?0.38). The resveratrol determination method, by using CdTe-CA QDs as fluorescence probe, was simple, rapid, inexpensive, and sensitive.     

Extraction and Preconcentration of Some Triazole Pesticides in Grape Juice by Salting Out Homogeneous Liquid–Liquid Extraction in a Narrow-Bore Tube Prior to Their Determination by Gas Chromatography–Flame Ionization Detection

A fast and simple extraction and preconcentration method for some triazole pesticides has been developed using a homogeneous liquid–liquid extraction method performed in a narrow-bore tube. The extraction is based on phase separation of a water-miscible organic solvent from aqueous solution in the presence of a salting out agent. In this work, the homogeneous solution of water and acetonitrile (water-soluble extraction solvent) was broken by addition of 30 %, w/v, sodium chloride (salting out agent). After sonication, a small volume of acetonitrile was collected on top of the tube and the extracted analytes in the collected phase were determined by gas chromatography–flame ionization detection. The effect of various experimental parameters including kind and volume of the water-soluble organic solvent, amount of salt, length and diameter of tube, and pH of sample solution was investigated. Under the optimum conditions, calibration graphs were linear over the range of 3–5,000 μg L^?1. Relative standard deviations were less than 5.4 % for six repeated determinations (C?=?100 μg L^?1). Furthermore, the limits of detection (S/N?=?3) and quantification (S/N?=?10) were obtained in the ranges of 0.60–4.8 and 1.9–16 μg L^?1, respectively. This method is very simple and rapid, requiring less than 10 min for sample preparation. It has been successfully utilized for the analysis of triazole pesticides in the grape juice samples.     

A Rapid Quantification of β-Carotene in Fruits and Vegetables by Dispersive Liquid–Liquid Microextraction Coupled with UV–Vis Spectrophotometry: Optimized by Response Surface Methodology

A simple, fast, and efficient method consisted of optimized dispersive liquid–liquid microextraction (DLLME) followed by UV–vis spectrophotometry was developed for determination of β-carotene in fruits and vegetables. Chloroform and methanol were chosen as extraction and disperser solvents, respectively. The extraction process was optimized using a central composite design (CCD) with the optimum points of 115 μL for volume of extraction solvent and 6.5 % (w/v) for salt concentration. Under the optimal conditions, the relative standard deviation (RSD, C?=?500 μg L^?1, n?=?5), limit of detection (LOD), linear dynamic range (LDR), and coefficient of determination (R²) were 1.08 %, 2 μg L^?1, 50–1,500 μg L^?1, and 0.991, respectively. The present method consisted of a simple and fast sample preparation procedure without any antioxidant addition, saponification, and purification was used.     

Simultaneous Determination of Nitrate and Nitrite in Fish Products with Improved Sensitivity by Sample Stacking-Capillary Electrophoresis

A fast and reproducible method for the simultaneous determination of nitrate and nitrite ions in canned fish samples by capillary zone electrophoresis has been developed. The sensitivity of the method was increased by applying a sample stacking technique. Optimal separation conditions were selected as 30 mmol L^?1 formic acid and 30 mmol L^?1 sodium sulfate at a pH of 4.0. The separation of nitrate and nitrite ions was achieved within 2.5 min. The limits of detection obtained at a signal-to-noise ratio of 3 for nitrate and nitrite were 0.55 and 0.82 μmol L^?1, while the relative standard deviations of intra-day corrected peak areas were 0.99 and 2.74 %, respectively. Recovery values ranged between 88.7 and 104 % for both ions. The method was successfully applied to canned fish samples, namely tuna, mackerel and sardine.     

Development of a New Methodology for Indirect Determination of Nitrite,Nitrate, and Total Nitrite in the Selected Two Groups of Foods by Spectrophotometry

A new indirect preconcentration procedure was developed for micellar sensitive detection of trace nitrite as analyte by spectrophotometry. The method is based on ion association of triiodide ion, I₃ ^? or I₂, produced by the reaction of nitrite at low concentrations with excess iodide in presence of ion-pairing agent, Coomassie brilliant blue R 250 (CBB), and electrophilic attack of nitrosyl cation produced by disproportionation of nitrite depending on concentration to ion-pairing agent at higher concentrations than 5 μg L^?1 in acidic medium, and then extraction of the ion-pairing complex formed from aqueous solution into the micelles of Triton X-114 at sodium acetate medium. The calibration graphs were rectilinear in the range of 0.5–5 and 5–200 μg L^?1 with increasing and decreasing slopes in micellar phase, respectively. The detection and quantification limits of the method were 0.15 and 0.50 μg L^?1, and the precision (as RSD) for five replicate measurements of different concentrations of nitrite was in the range of 2.3–4.8%. The average recoveries of spiked nitrite residues ranged from 97 to 104%. The method is free of matrix interferences and successfully applied to the indirect determination of nitrite, nitrate, and total nitrite in selected two groups of foods. Also, the accuracy was validated by analysis of a certified reference material as well as recoveries of spiked samples. The objective of the study is to provide an alternate economical method to determine the contents of nitrite, nitrate, and total nitrite after homogenization, extraction, reduction, and preconcentration from sample matrix.

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Graphical Abstract

     

Development of an ELISA and Immunochromatographic Assay for Tetracycline,Oxytetracycline, and Chlortetracycline Residues in Milk and Honey Based on the Class-Specific Monoclonal Antibody

A sensitive class-specific monoclonal antibody against tetracyclines (TCs) was generated and used to develop an enzyme-linked immunosorbent assay (ELISA) and an immunochromatographic assay for TC, oxytetracycline (OTC), and chlortetracycline (CTC) detection in milk and honey samples. The dynamic range of detection for TC in ELISA was 0.26–2.00 μg L^?1 with an IC₅₀ of 0.72 μg L^?1. The IC₅₀ value of OTC and CTC was 3.2 and 6.4 μg L^?1, respectively. The recovery of TC, OTC, and CTC in milk samples was 82–102, 91–105, and 90–101 %, respectively, and 88–101, 89–101, and 89–95 % in honey samples, respectively. In the immunochromatographic assay, the cutoff values for TC, OTC, and CTC were 15, 15, and 50 μg L^?1 in milk, respectively, and 40, 40, and 40 μg L^?1 in honey, respectively. The results revealed that ELISA and the immunochromatographic assay can be applied for the rapid and sensitive detection of TC, OTC, and CTC in milk and honey samples.     

Selective Determination of Selenium in Garlic, Mushroom and Water Samples by Chemically Modified Mesoporous Silica Solid Phase Coupled with ICP-OES

A selective sorbent for solid phase extraction (SPE), based on a chemically modified mesoporous silica (SBA-15), followed by inductively coupled plasma-optical emission spectrometry was used for extraction, preconcentration, and determination of selenium in water and food samples. The main parameters of SPE including pH, amount of mesoporous (solid phase), concentration of the eluent (desorption solvent), and equilibrium time were optimized by using a fractional central composite design (f-CCD). The optimum conditions were found to be 3.2 for pH, 21 mg for amount of the mesoporous, 1 mol l^?1 for eluent concentration, and 9 min for equilibrium time. Under the optimal conditions, the limit of detection (LOD) was 2.56 μg l^?1. The linear dynamic range (LDR) was 5–1,000 μg l^?1 with determination coefficient (R ²) of 0.999. Relative standard deviation (C?=?400 μg l^?1, n?=?5) was 3.84 %. The enrichment factor was 20. The maximum sorption capacity of the modified SBA-15 was 15 mg g^?1. The sorbent presented good stability, reusability, high adsorption capacity, and fast rate of equilibrium for sorption/desorption of selenium (IV) ions.     

Dual-Label Chemiluminescence Strategy for Multiplexed Immunoassay of 20 Fluoroquinolones, 15 β-Lactams, 15 Sulfonamides,and CAP in Milk

In this paper, a novel dual-label time-resolved chemiluminescent multiplexed immunoassay (DLTRC-MIA) based on the distinction of the kinetic characteristics of horseradish peroxidase (HRP) and alkaline phosphatase (ALP) with approximate estimation approach for simultaneous determination of 20 fluoroquinolones (FQs), 15 β-lactams, 15 sulfonamides (SAs), and chloramphenicol (CAP) in milk was developed. The strategy integrated a single-chain variable fragment–alkaline phosphatase fusion protein (scFv-ALP), a recombinant penicillin-binding protein (PBP) 2×*, a monoclonal antibody (MAb), and a polyclonal antibody (PAb) in one immunoassay and in a single well together to fulfill the simultaneous detection of 51 low-molecular weight contaminants (20 FQs, 15 β-lactams, 15 SAs, and CAP). The limits of detection for FQs, β-lactams, SAs, and CAP range from 0.29 μg L^?1 for ciprofloxacin (CIP) to 81.6 μg L^?1 for trovafloxacin (TRO), 0.27 μg L^?1 for ceftiofur (CEF) to 44.1 μg L^?1 for cephalexin (CEL), 0.089 μg L^?1 for sulfadimethoxine (SDM) to 2.7 μg L^?1 for sulfadiazine (SDZ), and 0.028 μg L^?1 for CAP, respectively. The results demonstrated that the detection limits of DLTRC-MIA meet the requirement of detection levels for 51 drug residues in milk, suitable for high-throughput screening of low-molecular weight contaminants.     

Determination of Hormones Residues in Milk by Gas Chromatography-Mass Spectrometry

The article presents the use of gas chromatography-mass spectrometry (GC-MS) technique in a method for the determination of 18 anabolic hormones from synthetic stilbenes, steroids and resorcylic acid lactones (RALs) groups in raw milk and milk powder. Sample preparation consisted of liquid-liquid extraction with diethyl ether and purification by solid phase extraction (SPE). Prior to instrumental analysis, the reaction of derivatisation with the heptafluorobutyric anhydride or N-methyl-N-trimethylsilyltrifluoroacetamide was performed. Method validation was carried out according to the required performance criteria of the Commission Decision 2002/657/EC. The apparent recovery of all analytes at 1 μg L^?1 (kg^?1) level was ranged between 70.4 and 119.4 % with the coefficients of variation values less than 30 %. The decision limits (CCα) and the detection capabilities (CCβ) were in the range of from 0.11 to 0.44 μg L^?1 (kg^?1) and from 0.19 to 0.75 μg L^?1 (kg^?1), respectively. The procedure has been accredited and successfully applied as a screening method for the presence of hormone residues in the study of commercial samples of milk.     

Effect of glycerol and dihydroxyacetone concentrations in the culture medium on the growth of acetic acid bacteria Gluconobacter oxydans ATCC 621

Bacteria of the species Gluconobacter oxydans are applied in the industrial production of dihydroxyacetone (DHA) via glycerol oxidation. The major problem of this biotransformation involves process inhibition by substrate and/or product. Improper initial concentration of glycerol and increasing DHA concentration may inhibit the metabolic activity of bacterial cells and impede further course of the reaction. An attempt was, therefore, undertaken in this study to determine which concentrations of glycerol (30, 50, 70, 100 g L^?1) and DHA (10–100 g L^?1) may inhibit the growth of acetic acid bacteria of G. oxydans ATCC 621 species. Cultures of this strain were run in the Bioscreen C MBR apparatus on experimental culture media with various initial concentrations of glycerol and DHA. Analyses were also carried out to examine the impact of pH (5.0, 7.0, 8.0) of glycerol-containing culture media on cell growth of the analyzed strain G. oxydans. None of the applied substrate concentrations was inhibiting cellular divisions of G. oxydans bacteria. The initial glycerol concentrations that enabled rapid cellular divisions reached 50 g L^?1 in the medium with pH 5.0 (coefficient of specific growth rate μ = 0.0550) and 70 g L^?1 in the medium with pH 7.0 (μ = 0.0556). DHA was shown to inhibit the mitotic activity of G. oxydans bacteria even at low concentrations (20–30 g L^?1), whereas at the concentration of 70 g L^?1, it made cell divisions impossible. The applied pH values of the culture media did not inhibit the growth of G. oxydans strain.     

Simultaneous Determination of Multiclass Pesticides and Antibiotics in Honey Samples Based on Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

A simple, fast, and efficient method was developed for simultaneous determination of 79 pesticides and 13 antibiotics compounds of different chemical classes of pesticides and antibiotics in honey samples by ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). The sample preparation procedure includes homogenization with McIlvaine buffer 0.1 mol L^?1 (pH 4), followed by extraction with acetonitrile and cleanup with florisil, using dispersive solid phase extraction (d-SPE). The proposed method was validated with good results, such as linearity (r ²?>?0.9901), normality, and independence of the evaluated data, as well as recoveries between 70 and 120 % with relative standard deviation (RSD) <20 % for most of the compounds spiked from 0.1 to 200 μg kg^?1. The experimental method limits of detection and quantification were from 0.03 to 1.51 μg kg^?1 and from 0.1 to 5 μg kg^?1, respectively, for the pesticides. For the antibiotics, the decision limits (0.1 to 2 μg g^?1) and the detection capacity (0.12 to 2.81 μg g^?1) were below the maximum residue limits (MRLs) established for honey by the Brazilian and European legislation. The method was successfully applied to real samples from different botanical and geographic origins. From them, 44 % presented residues from 0.12 to 10 μg kg^?1 of one or more analytes. The proposed method combines the advantages of a quick sample preparation step with the selectivity and sensitivity of the UHPLC-MS/MS and proved to be suitable for routine analyses.     

Speciation Determination of Selenium in Seafood by High-Performance Ion-Exchange Chromatography-Hydride Generation-Atomic Fluorescence Spectrometry

A high-performance ion-exchange chromatography coupled with hydride generation-atomic fluorescence spectrometry (HPIEC-HG-AFS) method was developed for simultaneous speciation of selenium in seafood. Three selenium species including of selenocystine (Se-Cys), selenome-thionine (Se-Met), and selenite Se(IV) were separated on an anion-exchange column (PRP-X100) with eluent of 30 mM NH₄H₂PO₄ and methanol (39:1, v/v) in 10 min at the flow rate of 1.0 mL min^?1. Variables affecting the HG-AFS detection of selenium species were optimized. The optimum conditions found were the following: reducing agent, 2.0 % of KBH₄, and 5.0 % of HCl; lamp current, 90 mA; photomultiplier tube voltage, 280 V; flow rate of carrier gas, 300 mL min^?1; and shielding gas, 800 mL min^?1. Under the optimized conditions, the good linearity of calibration curves (R ²?>?0.999) between signal of fluorescence and concentration of selenium species was obtained in the range of detection limits (DLs), 80 μg L^?1, and the DLs of Se-Cys, Se-Met, Se(IV) were 1.66, 0.990, 1.10 μg L^?1, respectively. The repeatability of the method, expressed as relative standard deviation, was less than 5.0 % (n?=?10), and the average recoveries for spiked test were from 87.3 to 103 % for three analytes in real seafood samples. The developed HPIEC-HG-AFS method was successfully applied for the speciation of selenium in seafood samples.     

Dispersive Liquid-Liquid Microextraction for the Determination of Emerging <Emphasis Type="Italic">Fusarium</Emphasis> Mycotoxins in Water

A new method was optimized for the determination of emerging Fusarium mycotoxins enniatins (ENs) and beauvericin (BEA) in different types of water. Mycotoxin analysis was performed by dispersive liquid-liquid microextraction (DLLME) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Mycotoxins were efficiently extracted from water into carbon tetrachloride by DLLME technique using acetonitrile as disperser solvent. Detection limits were in the range of 0.06–0.17 μg L^?1. Quantification was performed using matrix-matched calibration curves over a linear range from limit of quantification (LOQ) to 200 μg L^?1. Acceptable recoveries were obtained between 78.5 and 100.1 % with relative standard deviations of <14 %. The proposed method may be advised as an easy, sensitive, and accurate method for determining emerging Fusarium mycotoxins in water. The method was successfully applied to the analysis of different kinds of water. No detectable levels were achieved in surface, ground, tap, and bottled water. Concentration levels up to 50 μg L^?1 were detected in cooking water related to the pasta cooking process.     

Application and Optimization of Microwave-Assisted Extraction and Dispersive Liquid–Liquid Microextraction Followed by High-Performance Liquid Chromatography for the Determination of Oleuropein and Hydroxytyrosol in Olive Pomace

In the present study, a new method based on microwave-assisted extraction and dispersive liquid–liquid microextraction (MAE–DLLME) followed by high-performance liquid chromatography (HPLC) was proposed for the separation and determination of oleuropein (Ole) and hydroxytyrosol (HyT) from olive pomace samples. The effective factors in the MAE–DLLME process such as microwave power, extraction time, the type and volume of extraction, and dispersive solvents were studied and optimized with the aid of response surface methodology (RSM) based on a central composite design (CCD) to obtain the best condition for Ole and HyT extraction. At the optimized conditions, parameter values were 220 W microwave power, 12 min extraction time, 60 μL extracting solvent, and 500 μL dispersive solvent. The calibration graphs of the proposed method were linear in the range of 10–500,000 μg L^?1, with the coefficient of determination (R²) higher than 0.99 for Ole and HyT. Repeatability of the method, described as the relative standard deviation (RSD), was 4.12–5.63% (n?=?6). The limits of detection were 35 and 20 μg L^?1 for Ole and HyT, respectively. The recoveries of these compounds in the spiked olive pomace sample were from 93 to 98%. The proposed method, MAE–DLLME–HPLC–UV, was an accurate, rapid, and reliable method when compared with previous methods.     

Simple and fast spectrophotometric determination of low levels of thiabendazole residues in fruit and vegetables after pre-concentration with ionic liquid phase microextraction

There is a great importance of monitoring thiabendazole (TBZ) residues in fruits and vegetables to ensure food safety. Therefore, a new ionic liquid (IL) phase microextraction method using IL, 1-butyl-3-methylimidazoliumhexafluorophosphate [C₄mim][PF₆], as extracting solvent is proposed for simple and fast determination of low levels of TBZ in fruits and vegetables by spectrophotometry. The method is based on selective complex formation of TBZ with Cu(II) ions in presence of PF₆^– as counter ion at pH 5.5, and then microextraction of the complex into the fine micro-drops of IL phase. After optimisation of variables affecting microextraction efficiency, the analytical parameters of the method were determined by calibration curves. The method exhibits a linear relationship (0.3–280 μg L^?1), low detection limit (0.1 μg L^?1), good intra- and inter-day precision (2.4–4.5% as RSD_r%, 2.1–5.6% as RSD_R%), good recovery (≥95.1–98.2%) and high sensitivity enhancement factor (150) by solvent-based calibration curve. It allows a detection limit of 0.24 μg L^?1 and a range of 0.8–250 μg L^?1 by the matrix-matched calibration curve. After validation, the method was successfully applied to the determination of TBZ residues with method quantification limits in fruit and vegetables of 2.0 and 2.5 µg kg^?1 with and without adding polyvinylpyrrolidone (PVP-15) solution. Recoveries range from 85.5% to 98.2% after spiking (10, 50 and 100 µg kg^?1, n: 3).     

Large-Volume Sample Staking of Rice Polyphenols Prior to Their Determination by Non-aqueous Capillary Electrophoresis

A rapid non-aqueous capillary electrophoretic (NACE) method for the separation of (?)-epicatechin (EPI), (+)-catechin (?CAT), kaempferol (KAE), quercetin (QUR), naringanin (NAR), ferulic acid (FA), and p-coumaric acid (p-CA) has been developed and applied to the determination of these compounds in different rice varieties. All seven compounds were separated on capillary of 50 μm?×?68 cm (60-cm effective length) using 20 mmol L^?1 borate buffer of pH 9.0 and 5 % acetonitrile in methanol. Large-volume sample stacking (LVSS) technique was optimized and used to preconcentrate non-detectable polyphenols of white polished rice. Rice extracts were concentrated on-line by LVSS prior to separation by non-aqueous capillary electrophoresis. An improvement of 10–55 times in detectability was achieved with injection at 50 mbar for 30 s followed by voltage inversion (?20 kV) for 5 s. Linear calibration range of 1–300 μg L^?1 and 0.01–60 μg L^?1 was observed for NACE and NACE-LVSS method respectively, with the detection limit of 0.33–2.0 and 0.006–0.19 μg L^?1. Good reproducibility with standard deviations of less than 5 % was achieved. Polyphenol contents of different rice varieties were determined using developed method.     

Optimization and Validation of a Method Without Alkaline Clean-Up for Patulin Analysis on Apple Puree Agar Medium (APAM) and Apple Products

A sensitive high-performance liquid chromatography-UV (HPLC-UV) method, based on the Association of Analytical Communities (AOAC) Official method 2000.02, was developed and validated for the high-throughput analysis of patulin in in vitro experiments on apple puree agar medium (APAM). The importance of repeating the ethyl acetate extraction step at liquid-liquid extraction (LLE) was examined, as well as the extent of patulin degradation during the sodium carbonate clean-up. In addition to this alkaline clean-up, the efficiency of using an Oasis HLB or C₁₈ cartridge as solid-phase extraction (SPE) clean-up was compared. This resulted in a two-step ethyl acetate LLE, followed by an Oasis HLB SPE clean-up, without alkaline clean-up conditions. The method was fully validated for APAM, cloudy apple juice, and apple puree. Average patulin recoveries at levels of 100, 500, and 1000 μg kg^?1 of APAM varied between 95 and 113 % over 3 independent days, with an interday precision (RSD_R) of 5 to 10 %. Recovery experiments carried out with the spiked apple juice (at 50 μg kg^?1) and apple puree (10 μg kg^?1) showed average recovery rates laying between 80–101 % (RSD_R?=?12 %) and 77–100 % (RSD_R?=?9 %), respectively. This method offered a detection limit of 3–4 μg kg^?1 and a quantification limit of 5–8 μg kg^?1 for APAM, apple juice, and puree.     

Development and Validation of a Sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Screening of Florfenicol and Thiamphenicol in Edible Animal Tissue and Feed

To monitor the illegal use of florfenicol (FF) and thiamphenicol (TAP) in edible animal tissue and feed, a sensitive monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) has been developed with simple sample preparation and cleanup. The obtained monoclonal antibody (5F4) that has isotype IgG1 showed an IC₅₀ value of 0.21 μg L^?1 for FF and 0.35 μg L^?1 for TAP, respectively. It did not exhibit measurable cross-reactivity with other antibiotics. The limits of detection (LODs) for FF and TAP in a muscle matrix ranged from 0.07 to 0.14 μg kg^?1 and in a feed matrix ranged from 2.9 to 5.2 μg kg^?1. The recoveries were 72.8 to 113.4 % with a coefficient of variation of less than 15 %. Good correlation between the ELISA and HPLC-MS/MS results in the tissues tested demonstrated the reliability of our ic-ELISA. This ELISA is a useful tool for screening FF and TAP in edible animal tissue and feed.     

Preconcentration and Simultaneous Analysis of Benzimidazole Anthelmintics in Milk Samples by Ultrasound-Assisted Surfactant-Enhanced Emulsification Microextraction and High-Performance Liquid Chromatography

The proposed ultrasound-assisted surfactant-enhanced emulsification microextraction (UASEME) coupled with high-performance liquid chromatography-photodiode array detection (HPLC-PDA) has been developed for the preconcentration and simultaneous analysis of five benzimidazole anthelmintics. Dichloromethane (extraction solvent) and Triton X-114 (emulsifier) was used for extraction of the target analytes. The parameters affecting the extraction efficiency were investigated and optimized. Under the optimum conditions, linearity was in the range from 10 to 150 μg?L^?1 with good coefficients of determination (R ²) higher than 0.994. Preconcentration factors were obtained up to 60, corresponding to limits of detection range of 1.8???3.6 μg?L^?1. Intra-day (n?=?5) and inter-day (n?=?4?×?3) precisions were obtained with relative standard deviation of retention time and peak area below 0.8 and 9.2 %, respectively. Good recoveries for the spiked target anthelmintics at different concentrations (e.g., 20, 50, and 100 μg?L^?1) of milk samples were obtained in 72.5–113.5 %. The results demonstrated that the proposed UASEME-HPLC-PDA can be used as an alternative powerful method for the simultaneous determination of the target analytes in milk samples.