NMR Based Metabolomics Comparison of Different Blood Sampling Techniques in Awake and Anesthetized Rats.

Hongying Du,Fuqiang Xu,Huili Liu,Liang Wu,Anne Manyande,Jie Wang,Yingfeng Zhang,Shuang Li,Huiling Guo

doi:10.3390/molecules24142542

Abstract

The composition of body fluids has become one of the most commonly used methods for diagnosing various diseases or monitoring the drug responses, especially in serum/plasma. It is therefore vital for investigators to find an appropriate way to collect blood samples from laboratory animals. This study compared blood samples collected from different sites using the NMR based metabolomics approach. Blood samples were collected from the saphenous vein (awake state), tail vein (awake and anesthetized states after administration of sevoflurane or pentobarbital) and the inferior thoracic vena cava (ITVC, anesthetized state). These approaches from the saphenous and tail veins have the potential to enable the collection of multiple samples, and the approach from ITVC is the best method for the collection of blood for the terminate state. The compositions of small molecules in the serum were determined using the 1H-NMR method, and the data were analyzed with traditional correlation analysis, principle component analysis (PCA) and OPLS-DA methods. The results showed that acute anesthesia significantly influenced the composition of serum in a very short period, such as the significant increase in glucose, and decrease in lactate. This indicates that it is better to obtain blood samples under the awake state. From the perspective of animal welfare and multiple sampling, the current study shows that the saphenous vein and tail vein are the best locations to collect multiple blood samples for a reduced risk of injury in the awake state. Furthermore, it is also suitable for investigating pharmacokinetics and the effects of drug intervention on animals.

Highlights

For clinical applications, the composition of body fluids has become a commonly used standard for diagnosing various diseases or monitoring of drug responses
The saphenous vein and tail vein blood collection methods have the minimum adverse effects on the animals; this approach could be selected as the best representative of peripheral blood samples for potential multiple samples collection in the same animal
The current study compared the metabolites in blood samples using various blood collection methods, including different bleeding sites, i.e., saphenous vein/tail vein/inferior thoracic vena cava (ITVC), awake/anesthetized rats, and sevoflurane/pentobarbital induced anesthesia

Summary

Introduction

The composition of body fluids has become a commonly used standard for diagnosing various diseases or monitoring of drug responses. There are several kinds of body fluids, such as urine [1], blood plasma [2], serum [3], cerebrospinal fluid [4] etc. Among these samples, blood/serum measurements are the cornerstone of clinical testing; numerous investigations into the analysis of blood serum composition exist. Molecules 2019, 24, 2542 many common sites for blood collection in rodents, such as the tail vein (easy for catheterization), retro-orbital sinus, facial vein, saphenous vein, heart or the inferior thoracic vena cava (ITVC) [5,6]. For the collection of multiple blood samples over a short period of time, the approaches of retro-orbital sinus, tail vein, or saphenous vein are appropriate

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Conclusion