Identification of Tylosin Photoreaction Products and Comparison of ELISA and HPLC Methods for Their Detection in Water

Abstract

Tylosin is a widely used macrolide antibiotic for therapeutics and growth promotion in swine, beef cattle, and poultry production. Through various routes such as manure application, emission, inappropriate disposal, etc., tylosin enters the environment. The fate of tylosin in the environment is not yet fully understood. In this study, two photoreaction products of tylosin in water were identified as isotylosin A alcohol (E,Z) and isotylosin A aldol (E,Z). Tylosin A, B, C, D, isotylosin A alcohol, and isotylosin A aldol were purified, and immunological cross-reactivities of these tylosin-related compounds were tested with a specificity of 26% for tylosin B, 19% for tylosin C, 106% for tylosin D, 121% for isotylosin A alcohol, and 46% for isotylosin A aldol, compared to 100% for tylosin A. Competitive direct enzyme-linked immunosorbent assay (ELISA) for tylosin detection in water was compared with a high-performance liquid chromatography (HPLC) method by analyzing the same water samples from a study of tylosin dissipation in water. ELISA kits detect the other tylosin-related compounds besides tylosin A, which can result in differences in tylosin determination in water. Disciplines Biochemistry | Entomology Identification of Tylosin Photoreaction Products and Comparison of ELISA and HPLC Methods for Their Detection in Water Department of Entomology and Department of Biochemistry and Biophysics, Iowa State University, Ames, IowaReceived July 28, 2007. Revised manuscript received November 16, 2007. Accepted December 6, 2007 Tylosin is a widely used macrolide antibiotic for therapeutics and growth promotion in swine, beef cattle, and poultry production.Through various routes such as manure application, emission, inappropriate disposal, etc., tylosin enters the environment. The fate of tylosin in the environment is not yet fully understood. In this study, two photoreaction products of tylosin in water were identified as isotylosin A alcohol (E,Z) and isotylosin A aldol (E,Z). Tylosin A, B, C, D, isotylosin A alcohol, and isotylosin A aldol were purified, and immunological cross-reactivities of these tylosin-related compounds were tested with a specificity of 26% for tylosin B, 19% for tylosin C, 106% for tylosin D, 121% for isotylosin A alcohol, and 46% for isotylosin A aldol, compared to 100% for tylosin A. Competitive direct enzyme-linked immunosorbent assay (ELISA) for tylosin detection in water was compared with a high-performance liquid chromatography (HPLC) method by analyzing the same water samples from a study of tylosin dissipation in water. ELISA kits detect the other tylosin-related compounds besides tylosin A, which can result in differences in tylosin determination in water.

Determination of tylosins A, B, C and D in bee larvae by liquid chromatography coupled to ion trap-tandem mass spectrometry

“…Likewise, the length of the last stage of the gradient was also modified to stabilize the baseline for the following analyses. Nevertheless, the overall chromatography run was slightly shorter than that used in the only work where tylosins A, B, C and D have been determined [23], although the matrix was different (water) and three more Table 1. compounds were analyzed.…”

“…Several studies and even reviews have been published on the use of LC-MS/MS or LC-MS to analyze tylosin in several matrices, like water [23][24][25][26], honey [27][28][29][30][31] and food matrices [32][33][34][35], but not in bee larvae. In most of these studies only tylosin A was analyzed, although tylosin B was also studied in one report [28], and tylosins A, B, C and D have been determined in honey [27] and water [23]. As there are no prior references to studies in bee larvae, we set out to define a procedure to prepare the samples and to optimize a sensitive LC-MS/MS (MSD-Trap) method [27] in order to detect the lowest amounts of tylosins in this matrix.…”

Impact of veterinary antibiotics in the environment

“…The tylosin enzyme immunoassay used in this study measures all four tylosin factors (A-D) and two tylosin degradation products (isotylosin A alcohol and aldol) (Hu et al, 2008). Standard curves were prepared using tylosin tartrate, which contains each of the four tylosin factors, with tylosin A making up >80% of the formulation.…”

Low-Frequency Capacitive Sensing for Environmental Monitoring of Water Pollution with Residual Antibiotics

“…HPLC method was compared with enzyme-linked immunosorbent assay (ELISA) for detection of Tylosin and Tylosin-related compounds in water [17]. As a result, HPLC detected a lower concentration Tylosin compound while ELISA detected most of the Tylosin-related compounds.…”

“…Therefore, novel real-time antibiotics sensing techniques are required. Tylosin (C 46 H 77 NO 17) is a fermentation product of Streptomyces fradiae and it is used against gram-positive bacteria and some of the gram-negative bacteria [4,5].…”