Determination of Narasin in edible Swedish chicken tissue using solid phase extraction and high-performance liquid chromatography

Abstract

Introduction:
Narasin is a feed additive for animals to treat the parasitic disease Coccidiosis. The antibiotic is deadly to humans when consumed in large amounts. Studies has shown that Narasin can transfer on Vancomycin resistant E. Faecium (VRE). Vancomycin is an antibiotic used to treat bacterial infections in humans. The resistance could be passed on to humans from the chicken and contribute to yet another resistance issue.

Background:
Many studies on Narasin in feed has been done but very little work has been done on chicken tissue and in particular Swedish chicken. Moreover, when working with tissues quite often a cell disrupter or tissumeizer is used. In this report a bead mill will be used and evaluated in terms of recovery and repeatability.

Aim(s):
The aim of this study is to determine the amount of Narasin in Swedish chicken tissue using a bead mill instead of a cell disrupter in the sample preparation step.

Methods:
Parts of the method was done according to Tracey L. Cash et al report with small deviations. A bead mill was utilized instead of a cell disrupter and a vortex instead of a mechanical shaker. Chicken was purchased from a local store in Malmo. The tissue was homogenized with a blender. The samples were vortexed, centrifuged and cleaned using solid phase extraction. The analyte was analyzed using reversed-phase liquid chromatography with post-column derivatization for selective detection at 520 nm.

Results:
The result showed a LOD on 0,13 ppm and LOQ 0.43 ppm. The amount of Narasin in Swedish chicken tissue couldn’t be detected with this method. Regarding the use of a bead mill on three spiked chicken tissue samples showed a mean recovery at 57% and a standard deviation on 5. The repeatability of the three spiked samples showed a value on 8.72 %.

Conclusion:
In conclusion Narasin couldn’t be detected in Swedish chicken tissue with this method. The usage of a bead mill resulted in an acceptable recovery in accordance with the report done by Tracey L. Cash et al, where the acceptable value of spiked samples should be between 60% and 110%.

Popular Abstract

Narasin detection in Swedish Chicken tissue
In our everyday life chicken is considered a basic food component. The chicken industry is, and have been expanding drastically, which has forced the industry to take actions to preserve the expansion. Different additives are given to the animals to promote the expanding industry. The main purpose with the additives is fast growth and healthy chickens. Narasin is a drug used to prevent the sever disease, Coccidiosis. The illness has devastating effect on animals and has death as consequence. However, Narasin is an antibiotic and has proven to preserve and spread Vancomycin resistant E. Faecium (VRE) in chickens. Vancomycin is an antibiotic used to treat human infections and is classified by World health organization, as a significant antimicrobial for medicine. Nevertheless, a major resistance has been documented in hospitals where Vancomycin is widely used. This makes Vancomycin resistant E. Faecium a severe issue worldwide. The fact that Narasin preserve the spread of VRE in chickens, which humans then consume is worrisome.

High amounts of Narasin have been proven to be very harmful for humans. An accidental consumption of high amounts of Monesin, which is chemically very similar to Narasin, left humans with symptoms as vomiting, muscle weakness, respiratory failure and eventually death. Hence, the European union has put up a regulation for the maximum residue limit of Narasin in chicken tissue, to control and protect humans and animals. This regulation is taken very serious and regular controls is conducted to preserve that. In Norway, a step further was taken and all use of Narasin was banned since 2016. Instead of feed additives a vaccination program was conducted.

The purpose of this study is to investigate the amount of Narasin in Swedish chicken tissue, but also to evaluate the use of a bead mill in the extraction procedure. A bead mill is used to ground solid materials into a finer component. The bead mill was used to grind the chicken tissue with a solvent. The purpose of this technique is to break down the cells so that intracellular fluid becomes available to be dissolved in added solvent. This procedure is usually not used to extract Narasin from tissue. Therefore, there are a knowledge gap that this report aimed to investigate.

The method that was performed followed a validated method for determining Narasin in chicken tissue. Chicken tissue was purchased and minced with a blender then bead milled and cleaned in a series of steps. To analyze the Narasin a liquid chromatography was used with a derivatization. Narasin can’t be detected with UV/Vis since the compound doesn’t absorb light in the visible region. For that reason, an additional compound is added that react with Narasin. The reaction produces a chemical that is detected. Three samples were spiked with a known concentration Narasin. This method is used to detect how much of the Narasin is lost in the clean-up procedure.

The result in this study showed that the amount of Narasin in Swedish chicken tissue is too low to be detected with the method used. Because of that, the amount couldn’t be detected. The use of bead mill showed a low recovery. Which means that a lot of the Narasin put was lost in the cleaning-up process. The amount of Narasin in Swedish chicken tissue couldn’t be detected with this method. It also couldn’t be determined if the amount proceeded the maximum residue limit sat by the European union.

• author: Fathi, Mona
• supervisor: Margareta Sandahl
• organization: Department of Chemistry
• course: KEMK10 20211
• year: 2021
• type: M2 - Bachelor Degree
• subject: Chemistry
• keywords: Ball mill, Chicken, Derivatization, HPLC, Narasin, Analytical chemistry
• language: English
• id: 9057881
• date added to LUP: 2021-06-28 14:02:50
• date last changed: 2021-06-28 14:02:50