Sulfadiazine Residues Research Articles

Comparing the effects of irradiation and ultrasound on enrofloxacin, oxytetracycline, and sulfadiazine residues in chicken meat

PurposeThis research aims to evaluate and compare the effects of irradiation and ultrasound treatments on residual amount of three widely detected antibiotics in chicken meat, in addition to colorimetry and weight loss investigation. MethodsBroiler chickens were administered with three common antibiotics including enrofloxacin, oxytetracycline, and sulfadiazine and slaughtered at the age of 42 days. Chicken breast meat containing antibiotic residues was separately treated by gamma irradiation (at doses of 3, 5, and 7 kGy) and ultrasound at 240 W and 40 kHz (for 2, 5, and 10 min) to assess the effects of the treatments on reduction of antibiotic residues using HPLC technique. Moreover, the impact of treatments on weight loss and color changes were evaluated. ResultsThe results showed that both treatments significantly reduced the antibiotic residues; however, irradiation at higher doses and ultrasound at longer durations were more effective. The highest reduction of antibiotic residue was obtained at 7 kGy radiation dose and 10 min sonication time which was assigned to enrofloxacin (86.1%) followed by oxytetracycline (85%) and sulfadiazine (70.3%). Irradiation increased the L* (lightness) value and decreased the a* (redness) and b* (yellowness) values in chicken, but ultrasound had mostly no significant effect on the L* and a* values. Furthermore, both irradiation and ultrasound caused weight loss in the samples, but in the irradiated samples it was more than the sonicated ones. ConclusionsGenerally, using ultrasound for 10 min can be recommended as a better method to reduce the antibiotic residues in chicken.

Development and Validation of Aptasensor Based on MnO2 for the Detection of Sulfadiazine Residues.

The monitoring of sulfadiazine (SDZ) is of great significance for food safety, environmental protection, and human health. In this study, a fluorescent aptasensor based on MnO2 and FAM-labeled SDZ aptamer (FAM-SDZ30-1) was developed for the sensitive and selective detection of SDZ in food and environmental samples. MnO2 nanosheets adsorbed rapidly to the aptamer through its electrostatic interaction with the base, providing the basis for an ultrasensitive SDZ detection. Molecular dynamics was used to explain the combination of SMZ1S and SMZ. This fluorescent aptasensor exhibited high sensitivity and selectivity with a limit of detection of 3.25 ng/mL and a linear range of 5-40 ng/mL. The recoveries ranged from 87.19% to 109.26% and the coefficients of variation ranged from 3.13% to 13.14%. In addition, the results of the aptasensor showed an excellent correlation with high-performance liquid chromatography (HPLC). Therefore, this aptasensor based on MnO2 is a potentially useful methodology for highly sensitive and selective detection of SDZ in foods and environments.

Prototype for rapid test devices to detect residues of sulfonamides in chicken carcasses from traditional breeders in Surabaya, Indonesia.

Sulfadiazine, one of the sulfonamide group's active compounds, is widely used for therapeutic production against several diseases. Veterinary drug residues can have a significant impact on human health conditions. This study aimed to develop a prototype of rapid test devices (RTDs) for detecting sulfadiazine residues on chicken carcasses based on the color indication. Seven samples of carcasses collected from traditional breeders in Surabaya-Indonesia were prepared and tested using RTDs. This sample represents the population considering that in the last report, the use of antibiotics was more than 40%, while the ability to monitor RTDs was estimated at 100. The standard color of purple by Hex code standard color or decimal code color was used to compare the positive samples. A light-emitting diode (LED) lamp was used to observe purple color. Analysis of sulfonamides resulting from RTDs was compared using a ultraviolet-visible spectrophotometer. Sulfonamides contamination levels of 50% and 100% were detected at concentrations of 0.472 μg/mL and 0.642 μg/mL, respectively. Sulfonamides contamination that was <0.395 μg/mL did not appear purple. The study's findings showed that RTDs can be used to detect sulfonamides residues at a limit of detection 0.5 mg/mL after a 45 min exposure to an LED operating at a wavelength of 980 nm (p < 0.05). The limitation of RTDs was not being able to monitor the presence of residues bound in fat samples. Rapid test devices can be developed for commonly monitoring devices due to the limited technology available in the market.

The Forms, Distribution, and Risk Assessment of Sulfonamide Antibiotics in the Manure-Soil-Vegetable System of Feedlot Livestock.

Manure, soil, and vegetable samples were collected from different-sized livestock farms in Xinxiang, China. The residues of sulfadiazine, sulfamonomethoxine, and sulfamethoxazole were analyzed by high-performance liquid chromatography. The results indicated that the concentration ranges of the three total sulfonamides in manure, soil, and vegetables were 10.13-566.23µgkg-1, 7.60-176.26µgkg-1, and 0-32.70µgkg-1, respectively. The mean concentrations were 219.71µgkg-1, 70.73µgkg-1, and 7.08µgkg-1 for manure, soil, and vegetables, respectively. The mean concentrations in soil were lower than the ecotoxic effect trigger value (100µgkg-1), indicating a low risk for organisms in soil. The concentrations of the three sulfonamides varied significantly in different types of vegetables and all were lower than the acceptable daily intake value (50µg (kg day)-1). However, the potential ecotoxicity and danger to human and animal health via accumulation of the antibiotic in the food chain cannot be ignored.

Stress responses and biological residues of sulfanilamide antibiotics in Arabidopsis thaliana.

Sulfonamides (SAs) are antibiotics widely used in clinical practice, livestock and poultry production, and the aquaculture industry. The compounds enter the soil environment largely through livestock and poultry manure application to farmland. SAs not only affect plant growth, but also pose a potential threat to human health through SA residues in plant tissues. In particular, sulfamethoxazole (SMZ) has been classified as a Category 3 carcinogen by the World Health Organization, and thus its soil ecological toxicity and possible health risks are of concern. Using A. thaliana as a model plant, stress responses and biological residues of sulfadiazine (SD), sulfametoxydiazine (SMD), and SMZ were investigated in the present study. Root length and aboveground plant biomass were significantly inhibited by the three types of SA, whereas lateral roots exposed to SMD grew vigorously. The contents of chlorophyll a and chlorophyll b and photosystem II maximum photochemical quantum yield declined with increase in drug concentration, which indicated that exposure to SAs affected photosynthesis and inhibited chlorophyll synthesis in A. thaliana. With increase in drug concentration, reactive oxygen species (ROS) accumulation in the leaves increased significantly. Activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) were activated at low SA concentrations, but increased lipid peroxidation occurred with increase in SA concentration. Of the three compounds, SMZ was the most toxic to A. thaliana, followed by SD, and SMD was the least toxic. The results indicated that the risk of SMD entering an organism through the food chain is greater than that for SMZ and SD.

Development of Nanozyme-Labeled Biomimetic Immunoassay for Determination of Sulfadiazine Residue in Foods

The excessive use and ingestion of residues of sulfonamides harm the human body and the environment. To meet the requirements of the maximum residue limit specified by the Ministry of Agriculture of China, a rapid detection method urgently needs to be developed. In the present study, a molecularly imprinted polymer (MIP) that could selectively recognize sulfadiazine (SDZ) was prepared using SDZ as the template molecule and methacrylic acid as the functional monomer. Using MIP as biomimetic antibody and Au@Pt@SiO2 nanozyme as a marker, a new biomimetic immunoassay was developed to detect sulfadiazine. Under the optimal conditions, the limit of detection (IC15) and sensitivity (IC50) of this method were 0.09 and 6.1 mg/L, respectively. To determine the accuracy of this method, honey and milk samples spiked with sulfadiazine were analyzed, with recoveries in the range of 70.8%-90.2%. The method was also used for the quantitative analysis of sulfadiazine residues in milk powder and milk samples, producing results which correlated well with those obtained by high-performance liquid chromatography.

Detection of antibacterial residues in milk by HPLC-DAD and microbial inhibitor tests

The objective of this study was to assess the detection sensitivity of four commercial microbial inhibition assays (MIAs) in comparison with the results by High Pressure Liquid Chromatography with Diode Array Detection (HPLC-DAD) for residues of three tetracyclines, trimethoprim and sulfadiazine in fortified raw milk samples. MIAs showed fairly good results at detecting sulfadiazine residues, whereas concerning tetracyclines and trimethoprim residues, most of the kits showed insufficient results. HPLC analysis revealed high recoveries of examined compounds with detection limits below their respective maximum residue levels (MRLs). The HPLC method was validated determining linearity, accuracy, precision, detection capability and decision limit. Subsequently, both methods were used to monitor several cow and sheep milk samples for the presence of antibacterial agents. Results obtained showed that MIAs could be successfully used for the analysis of different milk types. However, milk spoilage which was observed in some cases could decrease the sensitivity of the kits, a fact that does not seem to influence the effectiveness of HPLC-DAD method.&lt;br /&gt;&lt;br /&gt;

Development of water-compatible molecularly imprinted solid-phase extraction coupled with high performance liquid chromatography–tandem mass spectrometry for the detection of six sulfonamides in animal-derived foods

A water-compatible molecularly imprinted polymer (MIP) was prepared by combining reversible addition-fragmentation chain transfer (RAFT) with reflux precipitation polymerization (RPP) for solid-phase extraction (SPE) coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC–MS/MS) to detect six sulfonamides (SAs) in animal-derived foods and water samples. The SEM images indicated the spherical structure of MIP materials. In water medium, MIPs still performed good specific adsorption for SAs. Under the optimized detection conditions, the proposed method offered good linearity (R2 > 0.996) for the six SAs with relative standard deviation (RSD) for ten replicate extractions of 1.64%–4.68%, limits of detection (S/N = 3) of 0.02 μg L−1-0.1 μg L−1 and recoveries of 63.49%–115.72%. Besides, the whole extract process avoided organic solvent consumption, and trace sulfadiazine residues were found in the pork sample at similar concentration of 3.57 and 3.79 μg kg-1 using the MIP-SPE method and commercial Oasis HLB cartridge, respectively. It is confirmed the method could be applied to quantitative detect the trace residue sulfonamides in animal-derived samples and water samples.

Bioaccumulation of sulfadiazine and subsequent enzymatic activities in Chinese mitten crab (Eriocheir sinensis)

The bioaccumulation of sulfadiazine and subsequent enzymatic activities in Chinese mitten crab (Eriocheir sinensis) were studied in microcosms, by exposing to 50, 100, 500 and 1000ng/L of sulfadiazine for 44days. An effective method for extracting sulfadiazine in crab tissues was established by modifying the cleanup method after ultrasound extraction, with improved recoveries of 61.8%, 93.7% and 100.5% in gill, muscle and liver samples. The results showed that sulfadiazine residues were all <3ng/g dry weight in different tissues, and that sulfadiazine bioaccumulation in crab was not dose-dependent. A significantly negative correlation was observed between acetylcholinesterase activity and the residue concentration of sulfadiazine during exposure to 50ng/L and 1000ng/L, and between alkaline phosphatase and sulfadiazine residues in the 100ng/L exposure group in the gill, suggesting that the two enzymes played an important role in the metabolism of sulfadiazine in crab.

Determination of sulfadiazine in eggs using molecularly imprinted solid-phase extraction coupled with high-performance liquid chromatography.

The development of a simple and effective method for the isolation and purification of sulfadiazine residues in food of animal origin is of great significance since it is a great danger to human health. An off-line molecularly imprinted solid-phase extraction with high-performance liquid chromatography method was proposed for the selective pretreatment and determination of sulfadiazine in eggs, rapidly and effectively. The molecularly imprinted polymer was proved to have a homogeneous spherical structure and porous surface morphology with excellent adsorption capacity of 5258 μg/g for sulfadiazine. The newly established method showed a good linearity in the range of 0-200 μg/L, low limits of detection (0.06 μg/L), acceptable reproducibility (RSD, 2.60-5.03%, n = 3), and satisfactory relative recoveries (78.22-86.10%). It was demonstrated that the proposed molecularly imprinted solid-phase extraction with high-performance liquid chromatography method could be applied to determine sulfadiazine in eggs, which simplified the pretreatment procedure and improved the accuracy of the analysis process by reducing the loss of sulfadiazine in the fat-removing procedure compared with traditional methods. Molecularly imprinted solid-phase extraction with excellent selectivity and adsorption capacity is a simple, rapid, selective, and effective pretreatment method for the determination of sulfadiazine in egg samples.

Automated Determination of Sulfadiazine in Water, Fish Plasma and Muscle by HPLC with On‐Line Column‐Switching

Fast, cheap and simple HPLC methods have been gaining importance in the field of antibiotic residue analysis especially in the context of environmentally friendly and sustainable aquaculture. An effective column‐switching technique for HPLC using online extraction column was developed to eliminate the time consuming pre‐treatment procedures and to facilitate the high throughput analysis of sulfadiazine in fish plasma and muscle as well as in water samples. The standard curves for the determination of sulfadiazine residues from biological matrices and water have good linearity (r2 &gt; 0.996). The average recovery ranged from 92.2 to 118% with the relative standard deviations of 0.3–9.3%. Limits of detection were determined as 5.7 μg L−1 for water, 8.1 μg L−1 for plasma and 9.6 μg kg−1 for muscle, while limits of quantification were 18.9 μg L−1 for water, 26.9 μg L−1 for plasma and 31.9 μg kg−1 for muscle, respectively. Intra‐ and inter‐day precisions were 0.3–9.3 and 0.4–6.7%, respectively. With the retention time of 12 min and total analysis time less than half an hour, this method provides an effective online extraction process and good sensitivity in liquid chromatography, and can be applied for the determination of residual sulfadiazine in water, fish plasma and muscle at lower than maximum residue level of 100 μg kg−1 fish.

Determination of Sulfadiazine Residues in Pork by Molecular Imprinted Column Coupling with High Performance Liquid Chromatography

Abstract A method for the quantitative monitoring of sulfadiazine (SD) residues in pork was established by molecular imprinted column coupling with high performance liquid chromatography (HPLC). The molecular imprinted column was selected as an extraction device. To obtain the optimum extraction efficiency, several parameters related to the molecular imprinted column, including column solvent, flow-rate, eluent of the sample matrix and eluent volume, were investigated. The sample solution was directly injected into the device for the extraction after simple extraction. Under the optimum conditions, the relative standard deviations (RSD) was ≤ 6.1%, and the recoveries for SD were higher than 75.6%. In comparison with the AL-SPE column, the MIP-SPE column had good reusability and extraction efficiency. This method was successfully applied to the determination of sulfadiazine residues in pork.

Residues of sulfadiazine and doxycycline in egg matrices due to cross-contamination in the feed of laying hens and the possible correlation with physicochemical, pharmacokinetic and physiological parameters

In the poultry industry, the widespread use of veterinary drugs such as antimicrobial compounds may lead to the presence of residues in whole eggs, egg white and egg yolk. During this study, laying hens received experimental feed containing sulfadiazine or doxycycline at cross-contamination levels of 2.5%, 5% and 10% of the therapeutic concentration. Since the therapeutic dose is 250 mg kg−1 for both substances, cross-contamination concentrations in the feed of 6.25, 12.5 and 25 mg kg−1 were expected. Whole egg, egg white and egg yolk samples were collected during the treatment and depletion period and were analysed via liquid chromatography-tandem mass spectrometry. For both drugs, a plateau phase was reached within 3–5 days and residue concentrations were detected in all egg matrices. For the 10% cross-contamination group, residual sulfadiazine concentrations of 208, 299 and 60 µg kg−1 and residual doxycycline concentrations of 455, 332, 206 µg kg−1 were detected in whole egg, egg white and egg yolk on day 13 of the treatment period, respectively. Both sulfadiazine and doxycycline had higher concentrations in egg white than in egg yolk, but the egg white–egg yolk ratio was higher for sulfadiazine than for doxycycline. As neither drug is allowed in Belgium for use in laying hens, residues may pose food safety concerns.

Residues of sulfadiazine and doxycycline in broiler liver and muscle tissue due to cross-contamination of feed

Veterinary drugs, such as antimicrobial compounds, are widely used in poultry and may lead to the presence of residues in matrices of animal origin, such as muscle and liver tissue. In this study, broilers received an experimental feed containing sulfadiazine or doxycycline at cross-contamination levels of 2.5, 5 and 10% of the therapeutic dose in feed. Breast and thigh muscle and liver samples were collected during treatment and depletion period and analysed via liquid chromatography–tandem mass spectrometry (LC–MS/MS). Concentrations reached a plateau phase 3–5 days after the start of experimental feeding. A rapid depletion of residues was noted after withdrawal of the experimental feed. No significant differences in measured concentrations were observed between the various muscle types. Residue concentrations for some experimental groups; the 10% group of sulfadiazine and the 5 and 10% group of doxycycline, however, exceeded their corresponding maximum residue limits (MRLs).

Sequestration of Manure-Applied Sulfadiazine Residues in Soils

It is not the total but the (bio)accessible concentration of veterinary medicines that determines their toxicity in the environment. We elucidate the changes in (bio)accessibility of manure-applied sulfadiazine (SDZ) with increasing contact time in soil. Fattening pigs were medicated with 14C-labeled SDZ, and the contaminated manure (fresh and aged) was amended to 2 soil types (Cambisol, Luvisol) and incubated for 218 days at 10 degrees C in the dark. Antibiotic residues of different bioaccessibility were approached by sequential extractions with 0.01 M CaCl2 (CaCl2 fraction), methanol (MeOH fraction), and finally acetonitrile/water (residual fraction, microwave extraction at 150 degrees C). In each fraction, total radioactivity, SDZ, and its major metabolites were quantified. The results showed that both SDZ and,to a lesser extent 4-hydroxysulfadiazine (4-OH-SDZ) were rapidly reformed from N-acetylsulfadiazine (N-ac-SDZ) during the first 2-4 weeks after fresh manure application, i.e., the N-acetylated metabolite does not sequester in soil to a significant extent Yet, the water and methanol extractable SDZ and 4-OH-SDZ also dissipated rapidly (DT50 = 6.0-32 days) for the fresh manure treatment with similar rate constants for both soil types. In the residual fractions, however, the concentrations of both compounds increased with time. We conclude that the residual fraction comprises the sequestered pool of SDZ and its hydroxylated metabolite. There they are entrapped and may persist in soil for several years. Including the residual fraction into fate studies thus yields dissipation half-lives of SDZ which exceed those previously reported for sulfonamides by a factor of about 100.

Analysis of aged sulfadiazine residues in soils using microwave extraction and liquid chromatography tandem mass spectrometry

An efficient extraction of sulfadiazine residues from soils is difficult, as sulfadiazine is known to form quickly sequestering residues. The objective of this study was to optimize an exhaustive extraction for aged residues of sulfadiazine and its two major metabolites, N-acetylsulfadiazine and 4-hydroxysulfadiazine, from soil. For this purpose two representative used agricultural soils (Luvisol, Cambisol) were blended with manure derived from [(14)C]sulfadiazine-treated pigs and incubated at 10 degrees C in the laboratory. After different extraction tests with various solvent mixtures (two- to four-component mixtures with water, methanol, acetonitrile, acetone, and/or ethyl acetate), different pH values (pH 4 and 9), and extraction temperatures (up to 200 degrees C), soil extracts were measured by liquid scintillation counting and liquid chromatography coupled to tandem mass spectrometry. With respect to sulfadiazine yields, stability of soil extracts, and the amount of coextracted matrix, a microwave extraction of soil (15 min, 150 degrees C) using acetonitrile/water 1:4 (v/v) is the method of choice for the exhaustive extraction of aged sulfadiazine residues from soils.

Screening for antibiotic residues in the trout by the Four Plate test, Premi test and ELISA test

The effectiveness of various assay systems for the examination of trout was investigated. The Four Plate test (FPT), Premi test and ELISA test were used for determining sulfadiazine/trimethoprim residues in trout. According to the results of the Four Plate test, when trouts were fed with pellets containing antibiotics (sulfadiazine/trimethoprim), the inhibition zones increased according to duration of feed application. In the control group, no inhibition zones were detected. The concentration of the residues accumulated and reached a plateau after 5 days. The antibiotics were detectable in the same concentration as on day 10, but 5 days later (on day 15) they were no longer detectable in the trout by EEC FPT. In contrast to the FPT, 5 days after discontinuing the medicated diet, there were still residues detectable by the Premi test and ELISA test on day 15. According to the ELISA test results of the trout; from day 1 to day 10 the results of sulfadiazine residues were found to be 4.18, 8.02, 9.13, 20.10, 21.32, 20.52, 20.75, 20.99, 20.90, 20.32 ppb, respectively. On day 10, the antibiotics were detectable in the same concentration. On day 15, sulfadiazine residues were found to be 6.82 ppb, but 5 days later (on day 20) they were no longer detectable in the trout. EEC Four Plate method presented less sensitivity and effectiveness in the detection of sulfadiazine.

Chemical and biological characterization of non-extractable sulfonamide residues in soil

For sulfonamides, the formation of non-extractable residues has been identified by laboratory testing as the most relevant concentration determining process in manured soil. Therefore, the present study has been focused on the chemical and biological characterization of non-extractable residues of 14C-labeled sulfadiazine or sulfamethoxazole. In laboratory batch experiments, the test substances were spiked via standard solution or test slurry to microbially active soil samples. After incubation periods of up to 102 d, a sequential extraction technique was applied. Despite the exhaustive extraction procedure, sulfadiazine residues mainly remained non-extractable, indicating the high affinity to the soil matrix. The remobilization of non-extractable 14C-sulfadiazine residues was monitored in the activated sludge test and the Brassica rapa test. Only small amounts (<3%) were transferred into the extractable fractions and 0.1% was taken up by the plants. In the Lumbricus terrestris test A, the release of non-extractable 14C-sulfamethoxazole residues by the burrowing activity of the earthworms was investigated. The residues mainly remained non-extractable (96%). The L. terrestris test B was designed to study the immobilization of 14C-sulfamethoxazole in soil directly after the test slurry application. The mean uptake by earthworms was 1%. Extractable and non-extractable residues amounted to 5% and 93%, respectively. Consequently, the results of all tests confirmed the high affinity of the non-extractable sulfonamide residues to the soil matrix.

CARBON NANOTUBES USED IN ELECTROANALYSIS

The fabrication of the carboxyl-modified CNT electrode was described. The electroanalytical investigation of sulfadiazine has been conducted in alkaline aqueous solution at the CNT electrode by voltammetry. Highly reproducible and well-defined cyclic voltammograms were obtained for sulfadiazine with a very good signal to background (S/B) ratio. However, no fouling of the electrode was observed at the CNT electrode within the experimental period of several hours, which illustrated that the CNT electrode was much better than traditional electrodes. Meanwhile, the detection of trace sulfadiazine in milk was also conducted by cyclic voltammetry with satisfactory ratio of recovery, indicating that the nanotube electrode can be used in routine monitoring of sulfadiazine residues in food.

Simultaneous determination of sulfadiazine and trimethoprim in animal feed by liquid chromatography with UV and tandem mass spectrometric detection

A method for the simultaneous determination of sulfadiazine and trimethoprim in animal feed was developed and validated. The chromatography was performed both on a liquid chromatography–ultraviolet detection (LC–UV) and LC–tandem mass spectrometric detection (MS/MS) instrument, intended to be applied for two separate purposes. The LC–UV method was used to examine the quality requirements for medicated premixes incorporated into animal feedingstuffs. This comprised e.g. the homogeneity testing in medicated feed at the mg kg −1 level. The LC–MS/MS method was used to quantify residues of sulfadiazine and trimethoprim in unmedicated feed at the μg kg −1 level. Sulfadiazine and trimethoprim were extracted from animal feed with 50% methanol in water and with ethylacetate. Clean-up was performed on a benzenesulfonic acid SCX cartridge. The LC–UV analysis of the extracts was done on a Hypersil ODS column using a gradient of 0.5% acetic acid, 0.25% triethylamine and 5% methanol in water (A) and 5% methanol in acetonitrile (B). Detection was at 240 nm for trimethoprim and 270 nm for sulfadiazine. The same extracts were also injected for quantitation onto an ion-trap LC–MS n instrument, operating in the atmospheric pressure chemical ionization tandem MS mode (APCI–MS/MS). The elution was performed isocratically with 0.1% acetic acid in water and methanol. Both the LC–UV and LC–MS/MS methods were validated for their linearity, trueness, precision, LOD (limit of detection), LOQ (limit of quantification) and extraction recovery. All results fell within the ranges specified. Both methods were useful, but the LODs of the LC–MS/MS method were about 200-times lower than with UV detection, i.e. 0.052 mg kg −1 sulfadiazine and 0.022 mg kg −1 trimethoprim. Therefore, the LC–MS/MS method was applied for monitoring residues as a possible result from carry-over during feed production.