Determination Of Residues In Honey Research Articles

Validation of two UHPLC-MS/MS methods for fast and reliable determination of quinolone residues in honey

ABSTRACT Antibiotic usage for beekeeping, including quinolones, can lead to residues in honey and these residues usually result from the drugs used in the treatment of American or European foulbrood diseases. Residues in honey may cause allergic reactions as well as increased antibiotic resistance. Within this study, rapid and breakthrough analysis methods were developed using UHPLC-ESI-MS/MS and sample preparation processes were minimised. Practical, low cost, time-effective analysis was provided utilising ‘dilute & shoot’ methodology and a QuEChERS-based procedure has been developed alternatively for improved sensitivity. Recovery values were 85%-112% for the ‘dilute & shoot’ method, and 82%-117% for the modified QuEChERS method. For accuracy studies, RSD% values were between 0.7% and 13.4% for both methods in intra-day and inter-day precision studies. CCα (1.1 μg/kg and 0.6 μg/kg) and CCβ (1.2 μg/kg and 0.6 μg/kg) mean values were calculated for the ‘dilute & shoot’ and QuEChERS method, respectively. These novel methods achieved simultaneous quantification of 21 quinolones in 7 min with high specificity and were used to analyse 50 honey samples. Quinolone residues were found in samples, and enrofloxacin, danofloxacin, pipemidic acid, lomefloxacin, cinoxacin, and ciprofloxacin were quantified.

Determination of Tetracycline Residues in Honey from Tabora and Singida Regions Tanzania Produced Using Modern Beehives

A liquid chromatographic tandem mass spectrometric (HPLC MS/MS) method is proposed for identification and quantification of antibiotics residues and metabolites in honey and other matrices such as water, manure, soil, meat and sediments. This method was used to detect tetracycline derived residues in honey samples from Tabora and Singida regions, Tanzania. This honey was obtained from modern beehives. Three hundred honey samples were tested for the presence of tetracycline derivative residues. Of the examined samples 15% had these drug residues. Majority of samples contained residues to a proportion of 0.002–0.008 mg/kg. The findings show that tetracycline antibiotic residues are present in honey from Tabora and Singida regions, although in small concentrations, however, this can possibly influence the naturally occurring protective properties of this medicinal food. Also can lead to drug resistance to consumers. Prospective studies that simultaneously track both resistance genes and antibiotic residues will go far in resolving some of the gagging questions that cloud our understanding of antibiotic resistance dissemination.

Impact of ion-pairs for the determination of multiclass antimicrobials residues in honey by LC-MS/MS

ABSTRACTThis study examines the behaviour of volatile perfluorinated carboxylic acids used as ion-pairing reagents for the separation of multiclass antimicrobial residues in honey (tetracyclines, sulphonamides, macrolides, and aminoglycosides). Heptafluorobutanoic acid and pentafluoropentanoic acid were compared as mobile phase additives at concentrations of 0–20 mmol.L−1 using reversed-phase chromatography. Our objective was to find optimal conditions that enable the separation of antimicrobials with a minimum loss in sensitivity and efficiency. The influence of ion pairing on chromatographic performance was examined. Results showed that heptafluorobutanoic acid was able to separate aminoglycosides faster and more efficiently than pentafluoropentanoic acid, but at low concentrations of heptafluorobutanoic acid between 2.5 and 5 mM, tetracyclines showed poor chromatographic efficiency. A concentration of 10 mM was found to be optimal for all studied veterinary drugs. The method was then applied to determine multiclass antimicrobial residues in honey. Based on validation data, the mean recoveries of analytes ranged between 93% and 104%, and the intermediate precisions were <21%. The decision limit (CCα) and detection capability (CCβ) were in the ranges of 5–25 and 7–33 μg kg–1, respectively. The method was found to be suitable for use in analysis of veterinary drugs with a broader group of compounds found in food-producing animals.

Desarrollo y validación de un método multiresiduo para el análisis de plaguicidas en miel por UFLC-MS

Se desarrolló un método para la determinación de residuos de plaguicidas en miel de abejas mediante cromatografía líquida ultra rápida acoplada a espectrometría de masas (cuádruplo sencillo). Se partió del método QuEChERS, evaluando las siguientes variables: (i) cantidad de muestra, (ii) tipo de sal para el control de pH, (iii) valor de pH del buffer y (iv) mezclas de limpieza. Adicionalmente y con el propósito de demostrar la confiabilidad del método, se evaluaron diferentes parámetros como exactitud, límites de detección y cuantificación, linealidad y selectividad. Los resultados mostraron que, mediante las modificaciones realizadas, se logró obtener un método más selectivo y se mejoró la exactitud de 19 plaguicidas respecto al método QuEChERS original. Por otro lado, mediante el proceso de validación, se encontró que el método es apto para el análisis de 50 plaguicidas (de 56 que se emplearon). Se obtuvieron porcentajes de recuperación entre el 70 y 120% y coeficientes de variación inferiores al 15%.

Development of a Gas Chromatography - Tandem Mass Spectrometry Procedure for Determination of Pesticide Residues in Honey and Honeybee Samples

A new analytical procedure of determination of multiresidue of pesticides based on application of the modified QuEChERS approach at the step of sample preparation and technique GC-MS/MS has been developed and applied for studies the real honey and honeybee samples. Samples were collected from northern Poland (Pomerania). The proposed procedure enables for the determination of 34 and 30 pesticide residues respectively in honey and honeybee samples. The recovery of analytes was between 70 and 120% with relative standard deviation ≤ 20%. The limits of quantification were in the range of 2.3-27 ng/g and 2.2-29 ng/g respectively for honey and honeybee samples. The proposed procedure was proven to be a powerful, highly sensitive, and environmentally friendly analytical tool that requires minimal sample preparation.

Multiresidue Determination of Pesticide Residues in Honey by Modified QuEChERS Method and Gas Chromatography with Electron Capture Detection

Honey can present pesticide residues due to the contamination of bees during the collection of pollen and nectar or by treatment of hives. Thus, the determination of pesticide residues in honey is of great importance, despite the difficulty due to the complexity of the matrix. In this study a new method for the determination of pesticides from different chemical groups in honey was developed and validated. Honey samples were extracted by modified QuEChERS method and analyzed by gas chromatography with electron capture detection (GC-ECD). Recovery results, evaluated at three spike levels, were between 71 and 119% for most of the compounds, with relative standard deviation (RSD) < 20%. The proposed method enables the determination at limits of detection between 3 and 6 µg kg-1, combining effective extraction and clean-up steps with good sensitivity and selectivity, and was successfully applied to the analysis of commercial honey samples.

Oxytetracycline Residues in Honey Analyzed by Liquid Chromatography with UV Detection

Summary A liquid chromatography method with UV detection for determination of oxytetracycline (OTC) in honey has been developed. The samples were extracted with the solution of oxalic acid. The clean-up procedure was performed by solid phase extraction (SPE) using polymeric Strata X and carboxylic acid cartridges. Chromatographic separation was carried out on the Luna C8 analytical column with mobile phase consisting of acetonitrile-0.02 M oxalic acid. The method has been successfully validated according to the requirements of the European Decision 2002/657/EC and this method is used in routine control of oxytetracycline in honey samples. The limit of detection (LOD) and limit of quantification (LOQ) of the presented method were 10 and 12.5 μg/kg, respectively. The developed method has also been verified in quantitative determination of oxytetracycline residues in honey after experimental treatment with this product in bee colonies.

Rapid Determination of Nitroimidazole Residues in Honey by Liquid Chromatography/Tandem Mass Spectrometry

We developed a rapid and efficient means of determining residues of four nitroimidazoles-i.e., dimetridazole, ipronidazole, metronidazole, and ronidazole-and three hydrophilic metabolites- i.e., 2-hydroxymethyl-1-methyl-5-nitroimidazole, 1 -methyl-2-(2'-hydroxyisopropyl)-5-nitroimidazole, and 1-(2-hydroxyethyl)-2-hydroxymethyl-nitroimidazole--in honey. We applied a QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) procedure improved to suit a nitroimidazole analysis, which is fast (approximately 30 min) and uses less organic solvent. The procedure involves initial single-phase extraction of 5 g of honey with acetonitrile containing 1% acetic acid, followed by liquid-liquid partitioning involving the addition of 5 g sodium chloride, 1.5 g trisodium citrate dihydrate, and 4 g magnesium sulfate. Moreover, matrix from honey was reduced by an SPE method with an alumina-N cartridge. The samples were analyzed using LC/MS/MS. Chromatographic separation of these nitroimidazoles and metabolites was performed in the gradient mode on a pentafluorophenylpropyl-bonded silica column (150x2.0 mm, 3 pm particle size) at 40 degrees C. The mobile phase consisted of a 0.01% acetic acid solution and acetonitrile, and the flow rate was 0.2 mL/min. The method was validated using honey spiked with these nitroimidazoles from 0.1 to 0.5 microg/kg. The overall recovery of the seven nitroimidazoles ranged from 76.1 to 98.5%; intra- and interassay CV values were <9.5 and <14.2%, respectively. The LOQ ranged from 0.1 to 0.5 microg/kg. LC/MS/MS coupled with the QuEChERS method showed good potential as a method for determining nitroimidazole residues in honey.

ＬＣ／ＭＳ／ＭＳによるハチミツおよびローヤルゼリー中のクロラムフェニコールの分析

A sensitive and selective method using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was developed for the determination of chloramphenicol (CAP) in honey and royal jelly. Mass spectral acquisition was performed in the negative mode by applying multiple reaction monitoring. In LC separation, Mightyl RP-18GP and 10 mmol/L ammonium acetate-acetonitrile were used as the column and mobile phase, respectively. CAP in honey samples was diluted with water, while CAP in royal jelly was extracted with 1% metaphosphoric acid-methanol (4 : 6). The solutions were cleaned up with an Oasis HLB cartridge. The quantification limits of CAP in honey and royal jelly were 0.3 ng/g and 1.5 ng/g, respectively. The recoveries of CAP from both honey and royal jelly at the quantification limits were over 92%. Twenty honey products and seven royal jelly products were analyzed by the developed method. CAP was detected in one honey product at 0.6 ng/g and in six royal jelly products at the level of 1.5-17.8 ng/g. These results show that the developed method has satisfactory sensitivity selectivity and is useful for the determination of CAP residues in honey and royal jelly.

Determination of residues in honey after treatments with formic and oxalic acid under field conditions

Formic acid and oxalic acid field trials for control of Varroa destructor were carried out in autumn according to the Swiss prescriptions during three successive years in different apiaries in Switzerland. The following parameters were determined in honey that was harvested the year after treatment: formic acid, ox- alic acid and free acidity. The following range of values were found in honeys of untreated colonies: formic acid, from 17 to 284 mg/kg, n = 34; oxalic acid, from 11 to 119 mg/kg, n = 33. There was a small, but unproblematic increase in formic acid levels in comparison to the levels in the controls; average: 46 mg/kg, maximum: 139 mg/kg. No increase in formic acid was found with increasing number of treatment years. If emergency formic acid treatments were carried out in spring, the residue levels were much higher: average in- crease of 193 mg/kg, maximum 417 mg/kg. The oxalic acid content remained unchanged, even after two suc- cessive treatments during the same autumn. No rise of free acidity was encountered after a combined treatment with formic and oxalic acid during the three trial years. honey/residue/formic acid/oxalic acid/free acidity/Varroa destructor

Determination of Cymiazole Residues in Honey by Liquid Chromatography

A liquid chromatographic method is described for the determination of cymiazole residues in honey. This acaricide is determined on a reversed-phase (C18) column, with a CH3CN-0.001N HCl-NaCl mixture (950 mL + 50 mL + 0.3 g/L) as the mobile phase, and UV detection at 265 nm. Cymiazole is extracted with n-hexane from aqueous alkalinized (pH 9) honey solutions. No further cleanup of the honey extract was required before chromatographic analysis. Recoveries on control samples fortified with 0.01, 0.10, and 1.00 ppm cymiazole ranged from 92 to 102%. The limit of determination was 0.01 ppm.