Chloramphenicol Residues In Food Research Articles

A fluorescence polarization aptasensor coupled with polymerase chain reaction and streptavidin for chloramphenicol detection

The authors describe a fluorescence polarization (FP) aptasensor based on the polymerase chain reaction (PCR) and streptavidin as dual FP amplifiers to detect chloramphenicol residues in food. Briefly, label-free aptamer was incubated with chloramphenicol and the aptamer-chloramphenicol conjugate was used as a template. Subsequently, the FAM-labeled forward primer and biotin-labeled reverse primer were added for PCR to amplify the template and the FAM-labeled primer. The molecular weight of FAM-labeled primer increased rapidly and the corresponding FP also enhanced. Finally, with the introduction of streptavidin, the PCR products and streptavidin were combined with the biotin-streptavidin interactions, resulting in much larger molecular weight. Thus, a dual amplified FP signal was obtained. Under optimal conditions, we were able to achieve a wide linear detection range of 0.001–200 nM. In addition, the designed strategy was applied to detect chloramphenicol in honey samples with high accuracy. Moreover, the strategy can be easily extended to detect other small molecules by changing the corresponding aptamers, which provide a promising avenue for the detection of small molecules by FP.

Analysis of Chloramphenicol and Two Metabolites in Crab and Shrimp Following Waterborne Exposure

The use of chloramphenicol (CAP) in aquaculture products is banned in many countries, including the United States, due to human health issues. Very few depletion and metabolism studies of CAP in seafood have been performed. Current detection methods for CAP residues in food are directed toward the parent drug molecule, but rapid elimination following treatment suggests the need for an alternative marker residue. We identified, characterized, and determined the persistence of two CAP metabolites, CAP-base (CAP-B) and CAP-alcohol (CAP-OH), in crab and shrimp. Interday recoveries of CAP, CAP-B, and CAP-OH in muscle fortified ( n = 9) at levels of 0.15 to 0.60 ng/g ranged from 95 to 127% and 101 to 119% for crab and shrimp, respectively, with repeatability ranging from 4 to 19%. The limit of detection for CAP and metabolites in crab and shrimp ranged from 0.05 to 0.11 ng/g. We also monitored the depletion of CAP, CAP-B, and CAP-OH in crab following waterborne exposures. To our knowledge, we present the first CAP depletion and metabolite study following waterborne exposure in crabs, with the aim of identifying alternative marker residues.

Total determination of chloramphenicol residues in foods by liquid chromatography-tandem mass spectrometry.

A simple and sensitive analytical method for the determination of chloramphenicol (CAP) and chloramphenicol glucuronide (CAPG) residues in foods such as livestock products, seafood, honey and royal jelly was developed. The method comprises solvent extraction with methanol, enzymatic hydrolysis with β-glucuronidase and clean-up using a hydrophilic lipophilic balanced copolymer solid phase extraction column. To determine the optimal conditions for the complete enzymatic hydrolysis of CAPG, we examined the effect of enzyme concentration and incubation time on the hydrolysis. The detection of CAP using LC-MS/MS was optimized and determined by SRM. The developed method was validated using ten food products at a spiked level of 0.5μg/kg. The validation results show excellent recoveries (79-109%) and precision (<15%) for CAP and CAPG. The limit of quantification (S/N≥10) of the developed method was 0.5μg/kg. The proposed method would be useful for the regulatory monitoring of CAP and CAPG residues in foods.

Chloramphenicol stereoisomers need to be distinguished: consequences observed from a proficiency test

ABSTRACTAlthough the use of chloramphenicol (CAP) as a veterinary drug is banned in the European Union and many other countries, monitoring for CAP residues in food is routine. Positive detections are few, but taken extremely seriously. European Union laboratories analysing for CAP should validate methods according to European Commission Decision 2002/657/EC, must be accredited to ISO 17025, and will generally participate in proficiency testing (PT) schemes, such as those offered by the Food Analysis Performance Assessment Scheme (FAPAS®). The FAPAS PTs aim to cover a wide range of relevant matrices including honey, prawns, fish, milk and kidney. Test materials are prepared either by animal dosing studies or by spiking raw matrix. The most common method reported by FAPAS participants used to screen for CAP residues is LC-MS/MS, but ELISA kits are increasingly being used. A recent PT round highlighted that the result obtained might be correlated with the type of analytical method being employed. Follow-up investigations have demonstrated that some of these variations in data are a function of the different stereoisomeric forms of CAP. This paper discusses the implication of this research on method validation requirements and European Union legislation.

Highly selective solid-phase extraction sorbents for chloramphenicol determination in food and urine by ion mobility spectrometry

Different highly selective sorbents have been evaluated for the treatment of food and biological samples to determine chloramphenicol residues by ion mobility spectrometry (IMS). Combination of a selective solid-phase extraction (SPE) and dispersive liquid-liquid microextraction allowed a highly sensitive determination of chloramphenicol in water, milk, honey, and urine samples. The performance of selective SPE supports such as immunoaffinity chromatography (IAC) and molecular imprinted polymers (MIP) have been compared in terms of selectivity, sensitivity, trueness, precision, and reusability. Quantitative recoveries were obtained for chloramphenicol residues, ranging from 91 to 123% for water, from 99 to 120% for skimmed milk, and from 95 to 124% for urine using IAC-IMS and MIP-IMS methods. Quantitative recoveries (from 88 to 104%) were also achieved for honey samples using IAC-IMS, but low recoveries were obtained using MIP-IMS. The limit of quantification was set at 0.1μgL-1 which is lower than the minimum required performance limit established by the EU. The proposed methodology is a simple and cost affordable alternative to chromatography methods for the highly sensitive and selective analysis of chloramphenicol residues in food and urine. Graphical Abstract Scheme for chloramphenicol determination by selective solid-phase extraction and ion mobility spectrometry.

Rapid surface enhanced Raman scattering detection method for chloramphenicol residues

Chloramphenicol (CAP) is a widely used amide alcohol antibiotics, which has been banned from using in food producing animals in many countries. In this study, surface enhanced Raman scattering (SERS) coupled with gold colloidal nanoparticles was used for the rapid analysis of CAP. Density functional theory (DFT) calculations were conducted with Gaussian 03 at the B3LYP level using the 3-21G(d) and 6-31G(d) basis sets to analyze the assignment of vibrations. Affirmatively, the theoretical Raman spectrum of CAP was in complete agreement with the experimental spectrum. They both exhibited three strong peaks characteristic of CAP at 1104cm−1, 1344cm−1, 1596cm−1, which were used for rapid qualitative analysis of CAP residues in food samples. The use of SERS as a method for the measurements of CAP was explored by comparing use of different solvents, gold colloidal nanoparticles concentration and absorption time. The method of the detection limit was determined as 0.1μg/mL using optimum conditions. The Raman peak at 1344cm−1 was used as the index for quantitative analysis of CAP in food samples, with a linear correlation of R2=0.9802. Quantitative analysis of CAP residues in foods revealed that the SERS technique with gold colloidal nanoparticles was sensitive and of a good stability and linear correlation, and suited for rapid analysis of CAP residue in a variety of food samples.

Detection of chloramphenicol residue in bovine meat using Liquid Chromatography Mass Spectrometry

Chloramphenicol (CAP) is a broad spectrum antibiotic that has been banned in many countries due to its serius side effect to human. Detection of CAP residue in food has been determined to a minimum required performance limit (MRPL) of 0.3 ng/g. The purpose of this research was to conduct the analysis of CAP residue in bovine meat by using LCMS and to study the presence of CAP residue in marketed bovine meat samples. LC separation was done on a Shimpack column C18 with ammonium acetate 10 mM/water as mobile phase, and ESI-MS analysis in negative ion mode. The coefficient of determination, R2 = 0.9981 at concentration of 0.125, 0.25, 0.63, 1,00 and 2.00 ng/g. Recovery at three fortification levels (0.25, 0.50 and 1.00 ng/g) was in the range 77.5, 97.3 and 83.4%. The decision limit and the detection capability were 0.15 ng/g and 0.17 ng/g respectively. Analysis results of 52 marketed samples showed that CAP residue were detected in 9 samples in the concentration range of 0.14 to 2.70 ng/g and 6 among those positive samples were above the MRPL value. Therefore, it is important to increase the awareness and also to monitor regularly CAP residues in food originated from animal to provide safe food for the consumers. Key Words : Chloramphenicol, Residue, LCMS, Bovine Meat

Determination of Chloramphenicol Residues in Foods by ELISA and LC-MS/MS Coupled with Molecularly Imprinted Solid Phase Extraction

The combination of molecularly imprinted solid-phase extraction (MISPE) with ELISA and LC-MS/MS was developed for the detection of chloramphenicol (CAP) in honey samples. Significant recoveries of 99.1 ± 7.1 and 98.8 ± 8.2% were obtained for intra- and inter-assay determination by ELISA determination, respectively. The limit of detection of CAP was 0.034 μg kg−1 and the limit of quantification was 0.046 μg kg−1. Determination and validation of CAP by using LC-MS/MS were performed following the same extraction and purification process as for the ELISA. The results demonstrated that the CAP samples purified by using MISPE were simultaneously applicable to analysis by ELISA and LC-MS/MS.

Molecularly imprinted polymer microspheres for solid-phase extraction of chloramphenicol residues in foods

Preparation of molecularly imprinted polymer microspheres (MIPMs) for chloramphenicol (CAP) by aqueous suspension polymerization is reported for the first time in this study. The resulting MIPMs had the ability to specifically adsorb CAP, and the molecularly imprinted solid phase extraction (MISPE) based on the MIPMs was shown to be applicable for clean-up and preconcentration of trace CAP in milk and shrimp samples with high recoveries of 92.7% and 84.9%, respectively. Combined with MISPE, the conventional HPLC-UV analysis sensitivity for CAP in foods could be significantly increased.

Analysis and monitoring of chloramphenicol residues in food of animal origin in Slovenia from 1991 to 2000

To prevent the illegal use of chloramphenicol (CAP), regulatory control of its residues in food of animal origin is essential. In Slovenia, the monitoring of CAP residues for statutory purpose started in 1991. The results of a 10-year period are presented. CAP residues were determined by capillary gas chromatography (GC) with electron capture detection (ECD) using meta-CAP as an internal standard (ISTD). Before chromatographic determination, analytes were derivatized by silylation. Overall, CAP recovery, adjusted for ISTD, was for bovine muscle tissue and raw cow's milk (in the region of 2–10 µg kg-1) 89 and 102%, respectively, and for whole eggs, 87% (in the region of 1–10 µg kg-1). The use of meta-CAP improved significantly the precision of the method. The detection limit for CAP was 1 µg kg-1, which was sufficiently sensitive for routine use. A total of 1308 random samples of Slovenian origin were analysed from 1991 to 2000, covering all parts of the country. CAP was found only in one milk sample in 1997 at a concentration of 4.6 µg kg-1.