Metabolic Effects of Clenbuterol and Salbutamol on Pork Meat Studied Using Internal Extractive Electrospray Ionization Mass Spectrometry

Yipo Xiao,Liping Luo,Shaoxian Xie,Haiyan Lu,Tenggao Zhu,Meng Cui,Haiwei Gu,Hua Zhang

doi:10.1038/s41598-017-05496-6

Abstract

Direct mass spectrometry analysis of metabolic effects of clenbuterol and salbutamol on pork quality at the molecular level is incredibly beneficial for food regulations, public health and the development of new anti-obesity drugs. With internal extractive electrospray ionization mass spectrometry (iEESI-MS), nutrients including creatine, amino acids, L-carnitine, vitamin B6, carnosine and phosphatidylcholines in pork tissue were identified, without sample pretreatment, using collision-induced dissociation (CID) experiments and by comparison with authentic compounds. Furthermore, normal pork samples were clearly differentiated from pork samples with clenbuterol and salbutamol via principal component analysis (PCA). Correlation analysis performed on the spectral data revealed that the above-mentioned nutrients strongly correlated with pork quality, and the absolute intensity of phosphatidylcholines in normal pork was much higher than pork contaminated by clenbuterol and salbutamol. Our findings suggested that clenbuterol and salbutamol may render effects on the activity of carnitine acyltransferase I, hence the process that L-carnitine transports long-chain fatty acids into mitochondria and the formation of phosphatidylcholines might be affected. However, the underlying metabolic mechanisms of clenbuterol and salbutamol on carnitine acyltransferase I requires more comprehensive studies in future work.

Highlights

Many researches were mainly limited to the development of simple, rapid, and sensitive methods for determination β-agonists residues in various biological samples, and evaluations of β-agonists on animal growth performance
The majority of dominant peaks in both normal pork sample and pork contaminated by clenbuterol and salbutamol at mass range of m/z 50–500 were assigned as amino acids[20, 23, 24] (e.g., m/z 118 [Valine+H]+, m/z 147 [Lysine+H]+, m/z 156 [Histidine+H]+, m/z 175 [Arginine+H]+), sugars (m/z 219 [Glucose+K]+), alkaloids (m/z 104 [Choline]+), polyamine
While in iEESI-MS, extraction solution biased with high voltage is continuously infused into bulk tissue sample through the parallelly inserted capillary for the extraction of “internal” chemicals from a bulk tissue sample

Summary

Introduction

Many researches were mainly limited to the development of simple, rapid, and sensitive methods for determination β-agonists residues in various biological samples, and evaluations of β-agonists on animal growth performance. Correlation analysis performed on the spectral data revealed that some nutrients strongly correlated with pork quality, and the absolute intensities of phosphatidylcholines in normal pork samples were much higher than in the samples contaminated by clenbuterol and salbutamol. These findings suggested that clenbuterol and salbutamol may render effects on the activity of carnitine acyltransferase I, the formation of phosphatidylcholines might be affected. The underlying metabolic mechanisms of clenbuterol and salbutamol on carnitine acyltransferase I require more comprehensive studies in future work

Methods

Results

Conclusion