Determination of Eight Coccidiostats in Eggs by Liquid–Liquid Extraction–Solid-Phase Extraction and Liquid Chromatography–Tandem Mass Spectrometry

Abstract

A method for the simultaneous determination of robenidine, halofuginone, lasalocid, monensin, nigericin, salinomycin, narasin, and maduramicin residues in eggs by liquid chromatography–tandem mass spectrometry (LC–MS/MS) was developed. The sample preparation method used a combination of liquid–liquid extraction (LLE) and solid-phase extraction (SPE) technology to extract and purify these target compounds from eggs. The target compounds were separated by gradient elution using high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC). Tandem mass spectrometry was used to quantitatively and qualitatively analyze the target compounds via electrospray ionization (ESI+) and multiple reaction monitoring mode. The HPLC–MS/MS and UPLC–MS/MS methods were validated according to the requirements defined by the European Union and the Food and Drug Administration. The limits of detection and limits of quantification of the eight coccidiostats in eggs were 0.23–0.52 µg/kg and 0.82–1.73 µg/kg for HPLC–MS/MS, and 0.16-0.42 µg/kg and 0.81-1.25 µg/kg for UPLC–MS/MS, respectively. The eggs were spiked with four concentrations of the eight coccidiostats, and the HPLC–MS/MS and UPLC–MS/MS average recoveries were all higher than 71.69% and 72.26%, respectively. Compared with the HPLC–MS/MS method, utilizing UPLC–MS/MS had the advantages of low reagent consumption, a short detection time, and high recovery and precision. Finally, the HPLC–MS/MS and UPLC–MS/MS methods were successfully applied to detect eight coccidiostats in 40 eggs.

Retracted: A review on the determination of veterinary drug residues in food products

“…Compared with the traditional method (LLE), SPE could enhance the analyte'srecovery rate, preserve time and effort, divide the analyte from the interference more productively, have a simple operating procedure and reduce the need for sample pretreatment (Wang, Liu et al, 2020). Some popular SPE cartridges such as hybrid SPE cartridges (Piatkowska et al, 2016), EVOLUTE ABN SPE cartridges (Kaufmann et al, 2011), CNWBOND LC-C18 SPE cartridges (Wang, Liu, et al, 2020) and hydrophilic-lipophilic balance (HLB) SPE cartridges (Dasenaki et al, 2016;Dasenaki & Thomaidis, 2015;Han et al, 2015;Turnipseed et al, 2017;Xie et al, 2015) have been used to eliminate veterinary drugs from honey, animal tissues, shrimp, eggs, meat, among others. Wang, Liu, and his colleagues (2020) utilized CNWBOND LC-C18 SPE cartridges to collect eight types of COCs from egg models such as nigericin, maduramicin, salinomycin, robenidine, narasin, lasalocid, halofuginone and monensin.…”

“…Mixed SPE (ratio 1:30, v/w) (Piatkowska et al, 2016) Eggs from the chicken CNWBOND liquid chromatography-C18 (ratio of 6:150, v/w) (Wang, Liu, et al, 2020) Dairy products and tissue from fish OASIS the balance of hydro-lipophilic (ratio of 3:60, v/w) (Dasenaki & Thomaidis, 2015) Milk (Han et al, 2015) Bovine muscle OASIS the balance of hydro-lipophilic (ratio of 6:200, v/w) (Dasenaki et al, 2016) Items made from milk (Xie et al, 2015) Tissue from animals, honey and fish EVOLUTE ABN (ratio of 3:200, v/w) (Kaufmann et al, 2011) Shrimp, eel and fish OASIS PRIME the balance of hydrolipophilic (ratio of 6:200, v/w) (Turnipseed et al, 2017) determined around 50 medicine residues in products from the fishing industry. For the QuEChERS technology, a dispersive SPE (denoted as the d-SPE) method utilizing primary secondary amine (denoted as PSA) and octadecylsilane (denoted as C18) absorbents, were chosen to restrict the matrix interference in the course of mass spectrometry analysis.…”

Veterinary Drug Residues in Animal-Derived Foods: Sample Preparation and Analytical Methods

“…The principle underlying the selectivity of SPE is similar to that of LC. Compared with the traditional LLE method, SPE can improve the recovery rate of the analyte, separate the analyte from the interfering components more effectively, and reduce sample pre-treatment processing, making it simple in operation and saving time and effort [ 61 ]. Common SPE cartridges include CNWBOND LC-C18 SPE cartridges [ 61 ], EVOLUTE ABN SPE cartridges [ 62 ], hydrophilic-lipophilic balance (HLB) SPE cartridges [ 63 , 64 , 65 , 66 , 67 ], and hybrid SPE cartridges [ 68 ], which are used to extract veterinary medicines from meat, milk, eggs, honey, fish, shrimp, eel, and animal tissues.…”

“…Recently, Wang et al [ 61 ] applied CNWBOND LC-C18 SPE cartridges to extract eight kinds of COCs (robenidine, halofuginone, lasalocid, monensin, nigericin, salinomycin, narasin, and maduramicin) from egg samples. The CNWBOND LC-C18 SPE cartridge has unique selectivity, and the long carbon chain also exhibits strong non-polarity.…”

Comparison of Two Commercially Available Immunoassays for the Measurement of Bovine Cardiac Troponin I in Cattle With Induced Myocardial Injury

“…Because of the mechanism of interfering with the cation membrane transport, the ionophores can cause cell death by destabilizing cell membranes, particularly in skeletal and cardiac muscle ( 30 ). Traditional methods of diagnosis of ionophore concentrations, such as liquid chromatography/mass spectrometry ( 31 ), while sensitive, are ill-suited for a rapid or on-farm diagnosis. As such, testing for elevated cTnI concentrations could hasten time to diagnosis for monensin toxicity in cattle.…”