Cattle hair as a bioindicator for short- and medium-chain chlorinated paraffin monitoring on cattle farms.

Short- and medium-chain chlorinated paraffins (SCCPs and MCCPs) are hazardous industrial chemicals that tend to bioaccumulate in animal-derived foodstuffs through the food supply chain. However, the lack of reliable noninvasive bioindicators hinders the monitoring of farm animal exposure to CPs. In this study, 169 cattle hair samples were collected from beef cattle farms in six Chinese provinces, with further beef, feed, and soil samples being collected in Hebei province. Geographical differences in CP concentrations were observed in the hair samples, and CP concentrations in samples collected from Hebei province decreased in the following order: hair > feed > beef > soil. C10-11Cl6-7 and C14Cl7-8 were the predominant SCCPs and MCCPs, respectively, in all the hair, beef, feed, and soil samples. CP concentrations in hair samples significantly correlated with those in beef, feed, and soil samples, indicating that hair can be used as a bioindicator of cattle exposure to CPs. The possible health risks associated with exposure to CPs through beef consumption, especially for children and high-volume beef consumers, should be further investigated.

A preliminary study on short- and medium-chain chlorinated paraffins in duck farms: Concentrations, distribution, and dietary exposure risks.

Chlorinated paraffins (CPs) in poultry feed and the farm environment might bioaccumulate in poultry eggs. Unlike chickens, which are mostly raised in cages, ducks are commonly raised free range. This would expose ducks to CPs in the environment. However, information on the presence of CPs on duck farms is scarce. In the present study, samples of duck eggs, duck feathers, poultry feed, and soil were collected from 25 duck farms in South China. Forty-eight congener groups of short- and medium-chain CPs (SCCPs and MCCPs) were detected in the samples. Interestingly, relatively high concentrations of SCCPs and MCCPs were found in the duck feathers. The median concentrations of SCCPs and MCCPs in the duck eggs, feathers, feed and soil were: 46 and 18 ng/g wet weight, 2460 and 992 ng/g, 103 and 47 ng/g, and 24 and 10 ng/g dry weight, respectively. The dominant groups of SCCPs and MCCPs were C10Cl6-7 and C14Cl7-8, respectively. The close relationship between duck feathers and poultry feed indicated that the duck feathers might act as a bioindicator for the exposure of ducks to CPs. The margin of exposure approach was used to assess the health risk, with the results showing that the consumption of duck eggs posed a low risk to different age groups from exposure to SCCPs and MCCPs.

Multiresidue Determination of 26 Quinolones in Poultry Feathers Using UPLC-MS/MS and Their Application in Residue Monitoring.

As a non-traditional sample matrix, feather samples can be used to effectively monitor antibiotic addition and organismal residue levels in poultry feeding. Therefore, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to simultaneously determine the residue levels of 26 quinolones in poultry feathers. The feather samples were extracted by sonication with a 1% formic acid and acetonitrile mixture in a water bath at 50 °C for 30 min, purified by the adsorption of multiple matrix impurities, dried with nitrogen, redissolved, and analyzed by UPLC-MS/MS. The linearity, limit of detection (LOD), limit of quantification (LOQ), recovery and precision were calculated. The 26 antibiotics demonstrated good linearity in the linear range. The recoveries and coefficients of variation were 78.9-110% and <13.7% at standard spiked levels of 10, 100 and 200 µg/kg, respectively. The LOD and LOQ were 0.12-1.31 and 0.96-2.60 µg/kg, respectively. The method also successfully identified quinolone residues in 50 poultry feather samples. The results showed that quinolones can accumulate and stabilize for a certain period of time after transferring from the body to the feathers of poultry.

Quick and high-throughput quantification of 22 ß-agonists residues in animal-derived foods using enzymatic probe sonication.

A high demand exists in veterinary drug residual analysis for rapid, automatic and high-throughput analytical techniques that produce data simultaneous and faster. Here, we describe a combined automated solid-phase extraction (SPE) and enzymatic probe sonication (EPS), subsequently ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the quick extraction, automated clean-up as well as simultaneous quantification of 22 ß-agonists residues in animal-derived foods. Enhanced by the ultrasonic probe, only 2 min was needed for exhaustively extraction of ß-agonists from foods of animal-origin; whereas traditional enzymatic digestion methods require hours or even days, which making it more appropriate for high-throughput biomonitoring. Moreover, the clean-up and pre-concentration procedures were conducted on the automatic SPE, which allowed 36 samples being performed simultaneously within 30 min. The method was successfully applied for analyzing 56 animal-derived food samples, 53.6 % of which contained detectable levels of at least one kind of ß-agonists. Interestingly, both the detection rate and residual level of ß-agonists in the ruminants (bovine and sheep) were higher as compared with the swine. Concerning the real food samples analyzed, the findings of this study suggest that stricter measures should be adopted to control the illegal usage of ß-agonists on the farm animals, particularly for the ruminants.

Fipronil and its metabolites in chicken feather: residue analysis, depletion study, and application analysis of pollution sources in laying hens.

A method based on a multi-mechanism impurity adsorption and ultra-performance liquid chromatography-tandem mass spectrometry was established to detect fipronil and four of its metabolites in chicken feathers. This method was successfully applied to the depletion study of fipronil in feathers of laying hens. Fipronil and two metabolites were found in feathers during treatment. Fipronil concentrations in feathers increased during medication and then regularly decreased during withdrawal, and they were still detected on the 14th day after withdrawal. High residue concentrations were also present in feathers on day 23 of the experimental period. Pollution sources of fipronil can be inferred on the basis of the residue ratio of fipronil metabolites from different pollution modes. Result shows that feathers were an effective matrix for residue monitoring and risk analysis of fipronil in animals and the environment.

Simultaneous Determination of 11 Mycotoxins in Maize via Multiple-Impurity Adsorption Combined with Liquid Chromatography-Tandem Mass Spectrometry.

In this study, multiple-impurity adsorption purification (MIA) technologies and liquid chromatography−tandem mass spectrometry (LC-MS/MS) were used to establish a method for detecting 11 mycotoxins in maize. The conditions for mass spectrometry and MIA were optimized. Maize was extracted with 70% acetonitrile solution, enriched, and purified using MIA technologies, and then, analyzed via LC-MS/MS. The results showed that the linear correlation coefficients of the 11 mycotoxins were >0.99, the sample recoveries ranged from 77.5% to 98.4%, and the relative standard deviations were <15%. The validated method was applied to investigate actual samples, and the results showed that the main contaminating toxins in maize were aflatoxins (AFs), deoxynivalenol (DON), fumonisins (FBs), ochratoxin A (OTA), and zearalenone (ZEN). Additionally, simultaneous contamination by multiple toxins was common. The maximum detection values of the mycotoxins were 77.65, 1280.18, 200,212.41, 9.67, and 526.37 µg/kg for AFs, DON, FBs, OTA, and ZEN, respectively. The method is simple in pre-treatment, convenient in operation, and suitable for the simultaneous determination of 11 types of mycotoxins in maize.

Effect of Germination on the Avenanthramide Content of Oats and Their in Vitro Antisensitivity Activities.

In this study, a method, based on an ultraperformance liquid chromatography coupled with high-field quadrupole orbitrap high-resolution mass spectrometry (UHPLC-QE-HF-HRMS) platform, was established for the trace determination of three major avenanthramides (AVNs). The MS conditions for determining the AVNs were optimized, and the cracking methods of avenanthramides were analyzed. The linear range of the results and the correlation coefficient were 1−2000 µg/L and >0.996, respectively. Further, the established method was employed for the determination of the AVN contents of oats at different germination times, and the results indicated that the AVN contents of Zaohua and Bayou oats increased 19.26 and 6.09 times, respectively, after germination. The total AVN content of both oat varieties reached a maximum on the fifth day of germination (153.51 ± 4.08 and 126.30 ± 3.33 µg/g for the Zaohua and Bayou oats, respectively). Furthermore, this study investigated the antiallergic and antioxidant activities of the germinated oats via hyaluronidase inhibition and 2,2-diphenyl-1-picrylhydrazyl (DPPH)-scavenging assays. The antiallergic and DPPH-scavenging abilities of the ungerminated forms of both oat varieties were weaker. However, on the fifth day of germination, the inhibition rate of anthranilamide hyaluronidase reached 72.7% and 67.3% for the Zaohua and Bayou oat varieties, respectively. The antiallergic abilities of the oats increased significantly on the fifth day of germination in terms of their antiallergic capacities and DPPH clearance (82.67% and 77.64% for the Zaohua and Bayou oats, respectively), and the two indicators exhibited similar trends. These findings demonstrated that AVNs exhibit good antisensitivity and antioxidation properties, and the antisensitivity effect correlated positively with the AVN content.

Occurrence and risk assessment of five kinds of antimicrobial in mattress on swine farm use ectopic fermentation systems in Zhejiang Province.

Mattress is among the main products of ectopic fermentation system (EFS); however, the research on the data of antimicrobial residues in the mattress of EFS and risk assessments of mattress have not been conducted. This study involved a scale survey to assess the levels and distributions of 54 antimicrobial residues, including 4 tetracyclines, 19 quinolones, 22 sulfonamides, 3 amphenicols, and 6 macrolides in mattress on 12 swine farms that use ectopic fermentation systems (EFS) in Zhejiang Province. A total of 25 antimicrobials were detected in mattress, and the total residue amount of antimicrobial in mattress samples of each farm was 0.77-28.2 g/T. Chlortetracycline had the highest contribution rate, and the residue amount of antimicrobial in mattress is not entirely determined by the start-up time of EFS but is related to the use of feed containing antimicrobial, medication habits, the level of mattress management, and maintenance methods of EFS. The risk assessments of antimicrobial in the mattress were carried out. The results show that the risk of using mattress of EFS for soil is low.

Simultaneous Determination of 21 Sulfonamides in Poultry Eggs Using Ionic Liquid-Modified Molecularly Imprinted Polymer SPE and UPLC-MS/MS.

An ionic liquid-modified molecularly imprinted polymer (IL-MIP) composite with sulfamethazine as a template molecule and methyl acrylic acid and 1-aminopropyl-3-methylimidazolium bromide as functional monomers was successfully synthesized. The achieved IL-MIP was characterized and evaluated in detail and utilized in the extraction and cleanup of sulfonamides (SAs) in poultry egg samples. The results demonstrated that the IL-MIP possessed a broad reorganization toward SAs and could selectively adsorb 21 kinds of SA compounds. Furthermore, the solid-phase extraction column based on the IL-MIP was used in the extraction and cleanup of 21 SAs in eggs, and the confirmatory detection of SAs was performed using ultraperformance liquid chromatography−tandem mass spectrometry. Under optimum conditions, the limits of detection (LODs) for all SAs ranged from 0.1 ng·g−1 to 1.5 ng·g−1, and the LOD of this method was better than those of the existing methods. The recoveries of SA compounds spiked in egg samples ranged from 84.3% to 105.8%, with low relative standard deviations (<15%). The developed method based on the IL-MIP extraction and cleanup was successfully used in the detection of 21 SAs in more than 100 real poultry egg samples. The results indicated that the proposed method was suitable for detecting 21 SAs in poultry eggs.

[Metal-organic framework UiO-67-based enrichment and purification of progesterone residues in milk].

Progesterone functions as an endocrine-disrupting compound. Imitating endogenous hormones disrupt the animals' hormone levels. The potential hazard of progesterone in milk cannot be neglected. Thus, research has focused on establishing an efficient and convenient pretreatment and analytical approach. In this study, a metal-organic framework (MOF) material UiO-67 was prepared, which possessed a large specific surface area and excellent stability. It was employed to enrich and purify trace progesterones in a complex milk matrix as a filler to integrate the solid phase extraction column. An approach based on MOF was developed using ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). This approach could simultaneously determine seven kinds of progesterone residues in milk. The element spectra of UiO-67 were first measured and analyzed using X-ray photoelectron spectroscopy. The chemical interaction between UiO-67 and progesterone was proved by comparing the changes in binding energy and relative contents of functional groups, and the adsorption efficiency of 1 mg/L and 5 mg/L progesterones by UiO-67 was studied. The adsorption efficiencies of UiO-67 for 1 mg/L and 5 mg/L progesterones were 99.73%-99.95% and 88.87%-99.23%, respectively, according to the results. It proved the efficient adsorption of UiO-67 to progesterones and ensured that subsequent studies went smoothly. Furthermore, key parameters, such as the amount of sorbent, elution solvent type, and pH value, were examined and optimized to obtain optimal extraction recovery of the progesterones. Spiked concentrations of 50 µg/L were employed for extraction optimization. All experiments were performed three times. It also evaluated the matrix effect on mass spectrum signal of the progesterones. The optimized results showed that the seven progesterones could be satisfactorily recovered when the amount of adsorbent was 40 mg, pH value of the sample solution was 5, and elution solution was 5-mL acetone. Additionally, the matrix effect of progesterone in the milk sample was <20%. The matrix effect could be neglected using the aforementioned approach to extract and purify progesterones in milk. Finally, the seven progesterones showed good linearity between 1 and 100 µg/L under the optimized conditions, with linear correlation coefficients values >0.99. The limits of detection (LODs) ranged from 0.06 to 0.30 µg/L, and limits of quantification (LOQs) ranged from 0.19 to 1.0 µg/L, respectively. At various concentration levels of progesterones in milk, the recoveries were 87.10%-105.58%, with relative standard deviations of 2.66%-9.64%. Most importantly, the approach was successfully employed to determine progesterone levels in milk samples, with results in good agreement with the standard SN/T 1980-2007. The proposed approach had the advantages of high sensitivity and satisfactory accuracy compared with the reported pretreatment and detection approaches of progesterone in milk. Satisfactory experimental results can be obtained without the calibration by isotope inner standard. Meanwhile, considering the excellent performance of MOF materials in reducing matrix interference in complex samples, such the application of materials offers a new approach. It can be employed to enrich and detect hazards in a complex matrix in the future.

Enzymatic probe sonication for quick extraction of total bisphenols from animal-derived foods: Applicability to occurrence and exposure assessment.

A high demand exists in bisphenols (BPs) screening studies for quick, reliable and straightforward analytical methods that generate data faster and simultaneously. Herein, we describe a combination of enzymatic probe sonication (EPS) and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for quick extraction and simultaneous quantification of eight important BPs in animal-derived foods. Results obtained demonstrated that the ultrasonic probe power could not only enhance the enzymatic hydrolysis efficiency, but also accelerate the liquid-liquid extraction procedure. Under optimized EPS parameters, one sample could be exhaustively extracted within 120 s, as compared with 12 h needed for the conventional enzymatic extraction which is more suitable for high-throughput analysis. The method was successfully applied to analyze residual BPs in animal-derived foods collected from Beijing, China. Widespread occurrence of BPA, BPS, BPF, BPAF, BPP, and BPB were found, with detection frequencies of 65.2%, 42.4%, 33.7%, 29.4%, 28.3%, and 27.2%, respectively. The highest total concentration levels of BPs (sum of the eight BPs analyzed, ΣBPs) were found in chicken liver (mean 12.2 µg/kg), followed by swine liver (6.37 µg/kg), bovine muscle (3.24 µg/kg), egg (2.03 µg/kg), sheep muscle (2.03 µg/kg), chicken muscle (1.45 µg/kg), swine muscle (1.42 µg/kg), and milk (1.17 µg/kg). The estimated daily intake (EDI) of BPs, based on the mean and 95th percentile concentrations and daily food consumptions, was estimated to be 5.687 ng/kg bw/d and 22.71 ng/kg bw/d, respectively. The human health risk assessment in this work suggests that currently BPs do not pose significant risks to the consumers because the hazard index (HI) was <1.

Quantitative selenium speciation in feed by enzymatic probe sonication and ion chromatography-inductively coupled plasma mass spectrometry.

A rapid, sensitive and species preservative analytical method for the simultaneous determination of six selenium (Se) species has been developed. Enzymatic probe sonication (EPS) was investigated as a novel and alternative technology for the extraction of Se species from feed matrices and the results were compared with the conventional hot water extraction, enzymatic hydrolysis and sequential extraction. The critical parameters of EPS such as enzyme types, extraction time, temperature, ultrasonic power and sample/enzyme ratio were varied with control. The Se species were separated and quantitatively determined by ion chromatography-inductively coupled plasma mass spectrometry (IC-ICP-MS). Under current optimised conditions, six inorganic and organic Se species were completely separated within 15 min in a single chromatographic run. The spectral interferences from the argon plasma 40Ar2, 40Ar37Cl or 1H79Br were effectively removed by employing the kinetic energy discrimination (KED) mode. Quantitative extraction for total Se (>94.8%) and more than 89.0% for the sum of different Se chemical forms without species transformation were obtained in only 60 s by applying the EPS treatment using aqueous protease XIV. The limits of detection (LODs) and quantification (LOQs) for Se species were in the ranges of 0.21-0.56 µg kg-1 and 0.69-1.87 µg kg-1, respectively. The proposed method was successfully applied to the speciation of Se in several reference materials and feed samples collected from the markets and local farms.

Deposition, depletion, and potential bioaccumulation of bisphenol F in eggs of laying hens after consumption of contaminated feed.

Increasing concerns over bisphenol A (BPA) as an endocrine disrupting chemical (EDC) and its adverse effects on both humans and animals have led to the substitution by structural analogs, such as bisphenol F (BPF), in many application areas. Information regarding to the carry-over of this emerging chemical in farm animals is essential for legislation and risk assessment purposes. In this study, a large-scale number of animal experiments were designed to investigate the transfer of BPF from feed to eggs. One control and three experimental groups of laying hens (72 hens per group) were fed with basal diets and BPF-contaminated feed at concentration levels of 0.1, 0.5 and 2.5 mg kg-1, respectively, for two weeks. The hens were then fed with BPF-free diets for a further four weeks. Eggs were collected daily, and separated into egg yolk and white for BPF analysis. The effects of different levels of BPF exposure on laying performance followed a non-monotonic dose-response curve, since low level BPF (0.1 mg kg-1) exposure did increase the laying rate, mean egg weight and daily feed intake, while high level BPF (2.5 mg kg-1) exposure showed a decreasing trend. BPF residues were detected in both egg yolks and whole eggs after two days of administration, and plateau phase was achieved within 9-18 days. There are clear linear dose-response relationships between the plateau BPF concentrations in feed and eggs. The residue of BPF was found mainly in egg yolks with conjugated form and depleted slowly (still detected 21 days after feeding the BPF-free diet of the high level group). Mean carry-over rate of 0.59% BPF from feed to eggs was obtained. Compared with the carry-over rates of PCBs and dioxins, BPF showed a relatively minor trend of bioaccumulation in eggs. To the best of our knowledge, this is the first report on the deposition, depletion, and bioaccumulation study of bisphenols in farm animals. The quantity of data can therefore be helpful in the frame of risk assessment, especially for a comprehensive estimation of consumer exposure to the residues of bisphenols.

Trace analysis of bisphenol A and its analogues in eggs by ultra-performance liquid chromatography-tandem mass spectrometry.

Presence of bisphenol A (BPA) and its analogues in various environmental matrices have been well studied by now, but there is still an insufficient data on the occurrence of BPA analogues in animal-derived foodstuffs. Hence, this work describes a quick, sensitive and reliable analytical method for the simultaneous determination of eight BPA analogues in eggs, applying a solid-phase extraction procedure with PRiME HLB cartridges that combines isolation of the analytes from matrix and the sample clean-up on a single column. The limits of detection (LODs) and quantification (LOQs) for BPA analogues were in the ranges of 0.02-0.05 µg kg-1 and 0.1-0.2 µg kg-1, respectively. Average recoveries were ranged between 82.7% and 105.2%. Quantitative analysis was performed on 24 real samples, the results indicate that low levels of BPA, BPB and BPS were detected, and interestingly, over ten folds higher BPP contamination levels compared to BPA were found in two egg samples.

Short- and medium-chain chlorinated paraffins in plastic animal feed packaging and factors affect their migration into animal feed.

Chlorinated paraffins (CPs) are used as plasticizers and flame retardants in plastics. Plastic packaging containing CPs is widely used for storage of animal feed, which can become contaminated by CPs that migrate into the feed. In the present study, 31 commercial animal feed packaging samples made of polypropylene (PP) or polyethylene (PE) were collected from animal feed manufacturers in China. The mean concentrations of short-chain chlorinated paraffins (SCCPs) in the PP and PE samples were 60.0 and 54.5 µg/g, respectively. The medium-chain chlorinated paraffin (MCCP) mean concentrations in the PP and PE samples were 62.7 and 9.23 µg/g, respectively. The carbon congener group profiles of SCCPs and MCCPs in the samples were different. The dominant SCCP and MCCP chlorine congener groups in all the samples were Cl6-7 and Cl6-8, respectively. Time and temperature influenced the migration of CPs from packaging into animal feed. As the time or temperature increased, the CP concentrations in the animal feed increased but the congener group profiles of the SCCPs and MCCPs in the animal feed did not change. To reduce contamination of animal feed by CPs, it is necessary to restrict the use of CPs in animal feed packaging.

Multiresidue Determination of 27 Sulfonamides in Poultry Feathers and Its Application to a Sulfamethazine Pharmacokinetics Study on Laying Hen Feathers and Sulfonamide Residue Monitoring on Poultry Feathers.

A method for the simultaneous determination of 27 sulfonamides in poultry feathers using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established in this study. The samples were extracted using 0.1 mol/L HCl solutions in a 60 °C water bath for 2 h, purified using hydrophilic-lipophilic balance solid-phase extraction, nitrogen-dried, and then reconstituted for UPLC-MS/MS analysis, which was performed with a CSH-C18 column. Linearity, limit of detection, limit of quantification, recovery, and precision were calculated in accordance with Commission Decision 2002/657/EC. For linearity, all standard curves showed a standard coefficient greater than 0.99, and the recoveries and coefficient of variation were 89-115% and <20%, respectively. The limit of detection and limit of quantification were 0.2-5 and 0.5-20 ng/g, respectively. The method was successfully applied to sulfamethazine (SMZ) residue accumulation monitoring in laying hen feathers and sulfonamide residue monitoring on poultry feathers. SMZ residue accumulation in the laying hen feathers was studied after administration with 100 mg/kg of SMZ for 21 consecutive days. SMZ residues were still detected in feathers 14 days after drug administration and persisted for up to 85 days. Results from 42 poultry feather samples showed that the feather is a suitable medium to monitor the illegal use of sulfonamides in poultry production.

Application of enzymatic probe sonication for selenium speciation in animal feeds.

Enzymatic probe sonication (EPS) was investigated as a novel and alternative technology for the extraction of five Se species SeCys2, MeSeCys, Se (IV), SeMet and Se (VI) from animal feeds. The critical parameters of EPS such as enzyme types, ultrasonic power, sample/enzyme ratio, extraction time and temperature were carefully optimized. Under current conditions, one sample can be extracted in 60s, in contrast to several hours required by the conventional enzymatic methods Moreover, the extraction is performed in pure water, no further temperature control or pH adjustment is needed, and therefore the risk of species interconvertion is drastically reduced or inhibited. The Se species were separated and quantitatively determined by high-performance liquid chromatography-hydride generation atomic fluorescence spectrometry (HPLC-HG-AFS), which allows the separation of organic and inorganic Se species in a single chromatographic run. The newly developed method was successfully applied to the determination of Se species in 18 feed samples collected from markets and local farms. Concerning the feed samples studied, the results of this work suggest that stricter measures should be taken to control the Se-enriched feed supplements in terms of Se species.

Pseudo template molecularly imprinted polymer for determination of 14 kind of ß-agonists in animal urine by ultra-high-performance liquid chromatography-tandem mass spectrometry.

A pseudo template molecularly imprinted polymer (MIP) for selective extraction of ß-agonists was prepared using 4-hydroxyphenethyl alcohol as pseudo template molecule, methyl acrylic acid as functional monomer, and ethylene glycol dimethacrylate as cross-linker. Selectivity and adsorption capacity of prepared MIP were studied in detail. MIP selectively adsorbed 14 types of ß-agonists. Advantages of the prepared polymer include non-template leakage in purification of target analytes and cheap cost. MIP was used to purify ß-agonists in urine samples. The developed method was combined with ultra-performance liquid chromatography-tandem mass spectrometry. In optimum conditions, recoveries of the developed method ranged from 84.2% to 109.8% with low coefficients of variation at below 15%. Limits of detection and quantification for 14 target ß-agonists in urine samples reached below 0.05ng/mL and 0.1ng/mL, respectively. The developed method based on MIP enrichment and purification were successfully applied to analysis of target analytes in 50 actual urine samples.

Development of a subcritical water extraction approach for trace analysis of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in poultry tissues.

Subcritical water extraction was investigated as a novel and alternative technology for the separation of trace amounts of chloramphenicol, thiamphenicol, florfenicol and its major metabolite florfenicol amine from poultry tissues and its results were compared with those of conventional shaking extraction, ultrasonic extraction, and pressurized liquid extraction. Decreasing the polarity of water by successively increasing the extraction temperature from 50°C to 200°C at the moderate pressure enabled selective, highly effective extractions to be performed. Rapid quantification of the target compounds was carried out by ultra-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). The critical parameters of subcritical water extraction such as solvent modifier, temperature, pressure, extraction time, and static cycles were varied with control. The optimized extraction procedures using subcritical water as extraction solvent, were carried out on a pressurized liquid extractor operated at 150°C and 100bar, applying two static cycles for 3min. Average recoveries of the four analytes from fortified samples ranged between 86.8% and 101.5%, with relative standard deviations (RSDs) lower than 7.7%. The limits of detection (LODs) and quantification (LOQs) for the target compounds were in the ranges of 0.03-0.5µgkg(-1) and 0.1-2.0µgkg(-1), respectively. The proposed method is fast, sensitive, water-based thus more environmental acceptable, making it a suitable replacement for conventional organic solvent extraction in veterinary drug residue analysis.

Determination of ractopamine in animal hair: application to residue depletion in sheep and residue monitoring.

An ultra-performance liquid chromatography tandem mass spectrometry method to determine ractopamine (RAC) in hair samples from swine, sheep, and cattle was developed. The procedure to extract from incurred hair was optimized. The samples were extracted with 0.1mol/L HCl solution. The mixture was heated to 60°C in water bath for 4h. The extracts were purified by solid-phase extraction, dried under a stream of nitrogen, and then reconstituted in mobile phase for analysis, which was performed with a Waters BEH-C18 column. The limit of detection was 0.3ng/g, the limit of quantification was 1ng/g, the recoveries were between 87% and 105%, and the coefficient of variation was less than 15%. The depletion of RAC in hair was studied in healthy sheep after administration of RAC at 1000ng/g body weight for five consecutive days. The RAC residues were still detected with 82.5±7.2ng/g hair 27 days after drug administration. Five samples tested positive, in which the amount of RAC detected was 13-253ng/g from 569 hair samples collected from farms and slaughterhouses. These results show that animal hair is a suitable medium to monitor the illegal use of RAC in livestock production.