Development of metabolome extraction strategy for metabolite profiling of skin tissue

IF 3.5 3区 医学 Q2 ENDOCRINOLOGY & METABOLISM
Ting Bu, Sooah Kim
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引用次数: 0

Abstract

Introduction

Changes in skin phenotypic characteristics are based on skin tissue. The study of the metabolic changes in skin tissue can help understand the causes of skin diseases and identify effective therapeutic interventions.

Objectives

We aimed to establish and optimize a non-targeted skin metabolome extraction system for skin tissue metabolomics with high metabolite coverage, recovery, and reproducibility using gas chromatography/mass spectrometry.

Methods

The metabolites in skin tissues were extracted using eleven different extraction systems, which were designed using reagents with different polarities based on sequential solid-liquid extraction employing a two-step strategy and analyzed using gas chromatograph/mass spectrometry. The extraction efficiency of diverse solvents was evaluated by coefficient of variation (CV), multivariate analysis, metabolites coverage, and relative peak area analysis.

Results

We identified 119 metabolites and the metabolite profiles differed significantly between the eleven extraction systems. Metabolites with high abundances in the organic extraction systems, followed by aqueous extraction, were involved in the biosynthesis of unsaturated fatty acids, while metabolites with high abundances in the aqueous extraction systems, followed by organic extraction, were involved in amino sugar and nucleotide sugar metabolism, and glycerolipid metabolism. MeOH/chloroform-H2O and MeOH/H2O-chloroform were the extraction systems that yielded the highest number of metabolites, while MeOH/acetonitrile (ACN)-H2O and ACN/H2O-IPA exhibited superior metabolite recoveries.

Conclusion

Our results demonstrated that our research facilitates the selection of an appropriate metabolite extraction approach based on the experimental purpose for the metabolomics study of skin tissue.

Abstract Image

开发用于皮肤组织代谢物分析的代谢组提取策略
引言 皮肤表型特征的变化基于皮肤组织。研究皮肤组织中的代谢变化有助于了解皮肤疾病的病因,并确定有效的治疗干预措施。目的我们旨在建立并优化一种非靶向皮肤代谢组提取系统,利用气相色谱/质谱技术进行皮肤组织代谢组学研究,该系统具有高代谢物覆盖率、高回收率和高重现性。方法 使用 11 种不同的萃取系统萃取皮肤组织中的代谢物,这些萃取系统是基于两步法固液萃取顺序设计的,使用不同极性的试剂,并使用气相色谱/质谱法进行分析。通过变异系数(CV)、多元分析、代谢物覆盖率和相对峰面积分析评估了不同溶剂的萃取效率。在有机萃取系统中含量较高的代谢物参与了不饱和脂肪酸的生物合成,而在水提取系统中含量较高的代谢物参与了氨基糖、核苷酸糖和甘油脂的代谢,其次是水提取。MeOH/氯仿-H2O和MeOH/H2O-氯仿是获得代谢物数量最多的萃取体系,而MeOH/乙腈(ACN)-H2O和ACN/H2O-IPA则表现出更高的代谢物回收率。
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来源期刊
Metabolomics
Metabolomics 医学-内分泌学与代谢
CiteScore
6.60
自引率
2.80%
发文量
84
审稿时长
2 months
期刊介绍: Metabolomics publishes current research regarding the development of technology platforms for metabolomics. This includes, but is not limited to: metabolomic applications within man, including pre-clinical and clinical pharmacometabolomics for precision medicine metabolic profiling and fingerprinting metabolite target analysis metabolomic applications within animals, plants and microbes transcriptomics and proteomics in systems biology Metabolomics is an indispensable platform for researchers using new post-genomics approaches, to discover networks and interactions between metabolites, pharmaceuticals, SNPs, proteins and more. Its articles go beyond the genome and metabolome, by including original clinical study material together with big data from new emerging technologies.
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