Simultaneous extraction of metabolome and lipidome with methyl tert-butyl ether from a single small tissue sample for ultra-high performance liquid chromatography/mass spectrometry

Chen, Shili; Hoene, Miriam; Li, Jia; Li, Yanjie; Zhao, Xinjie; Häring, Hans‐Ulrich; Schleicher, Erwin; Weigert, Cora; Xu, Guowang; Lehmann, Rainer

doi:10.1016/j.chroma.2013.05.019

Cited by 190 publications

(151 citation statements)

References 38 publications

(44 reference statements)

Supporting

Mentioning

149

Contrasting

Order By: Relevance

“…The protocol had been validated in previous studies (18,19). For each specimen, 10 mg of wet tissue was weighed and placed into a 2 ml centrifuge tube.…”

Section: Methodsmentioning

confidence: 99%

Integration of Metabolomics and Transcriptomics Reveals Major Metabolic Pathways and Potential Biomarker Involved in Prostate Cancer

Ren

Shao

Zhao

et al. 2016

Molecular & Cellular Proteomics

Self Cite

166

130

View full text Add to dashboard Cite

Prostate cancer is a highly prevalent tumor affecting millions of men worldwide, but poor understanding of its pathogenesis has limited effective clinical management of patients. In addition to transcriptional profiling or transcriptomics, metabolomics is being increasingly utilized to discover key molecular changes underlying tumorigenesis. In this study, we integrated transcriptomics and metabolomics to analyze 25 paired human prostate cancer tissues and adjacent noncancerous tissues, followed by further validation of our findings in an additional cohort of 51 prostate cancer patients and 16 benign prostatic hyperplasia patients. We found several altered pathways aberrantly expressed at both metabolic and transcriptional levels, including cysteine and methionine metabolism, nicotinamide adenine dinucleotide metabolism, and hexosamine biosynthesis. Additionally, the metabolite sphingosine demonstrated high specificity and sensitivity for distinguishing prostate cancer from benign prostatic hyperplasia, particularly for patients with low prostate specific antigen level (0 -10 ng/ml). We also found impaired sphingosine-1-phosphate receptor 2 signaling, downstream of sphingosine, representing a loss of tumor suppressor gene and a potential key oncogenic pathway for therapeutic targeting. By integrating metabolomics and transcriptomics, we have provided both a broad picture of the molecular perturbations underlying prostate cancer and a preliminary study of a novel metabolic signature, which may help to discriminate prostate cancer from normal tissue and benign prostatic hyperplasia. Molecular & Cellular Proteomics

show abstract

“…The protocol had been validated in previous studies (18,19). For each specimen, 10 mg of wet tissue was weighed and placed into a 2 ml centrifuge tube.…”

Section: Methodsmentioning

confidence: 99%

Integration of Metabolomics and Transcriptomics Reveals Major Metabolic Pathways and Potential Biomarker Involved in Prostate Cancer

Ren

Shao

Zhao

et al. 2016

Molecular & Cellular Proteomics

Self Cite

166

130

View full text Add to dashboard Cite

show abstract

“…A long run time is also a disadvantage for two-dimensional LC [HILIC combined with reversed-phase (RP) LC] ( 20 ). RP chromatography (RP-LC) has been widely applied for lipidomic analyses using different stationary phases [e.g., diphenyl ( 21 ), C 8 (22)(23)(24), and C 18 ( 23,(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)]. For several years, our group has used high strenght silica (HSS) T3 columns (C 18 material) for our lipidomic analyses ( 33 ).…”

Section: Uplc Analysismentioning

confidence: 99%

Enhanced lipid isomer separation in human plasma using reversed-phase UPLC with ion-mobility/high-resolution MS detection

Damen

Isaac

Langridge

et al. 2014

Journal of Lipid Research

113

105

View full text Add to dashboard Cite

This article is available online at http://www.jlr.orgLipids are the building blocks of all cell membranes and are thus essential for various biological functions varying from membrane traffi cking to signal transduction. Disorders in lipid metabolism play a key role in various diseases including cardiovascular disease, cancer, diabetes mellitus, and infl ammation ( 1, 2 ). Lipids have been classifi ed into eight categories (fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, sterol lipids, prenol lipids, saccharolipids, and polyketides) by the International Committee for the Classifi cation and Nomenclature of Lipids in conjunction with the LIPID MAPS consortium, based on their chemically distinct functional backbones and hydrophobic nature ( 3, 4 ).As the study of lipids attracts more and more attention, lipidomics, as a branch of metabolomics, is a quickly emerging fi eld. It has been defi ned as "the full characterization of lipid molecular species and of their biological roles with respect to expression of proteins involved in lipid metabolism and function, including gene regulation" ( 5 ).In order to achieve the goal of full characterization, different MS-based approaches have been described. Some groups describe shotgun lipidomics using direct infusion into the mass spectrometer ( 6-11 ). Direct infusion MS has the advantage of short analysis times and consuming very small amounts of sample. However, structural elucidation of lipid isomers is not possible. In order to elucidate structures Abstract An ultraperformance LC (UPLC) method for the separation of different lipid molecular species and lipid isomers using a stationary phase incorporating charged surface hybrid (CSH) technology is described. The resulting enhanced separation possibilities of the method are demonstrated using standards and human plasma extracts. Lipids were extracted from human plasma samples with the Bligh and Dyer method. Separation of lipids was achieved on a 100 × 2.1 mm inner diameter CSH C 18 column using gradient elution with aqueous-acetonitrile-isopropanol mobile phases containing 10 mM ammonium formate/0.1% formic acid buffers at a fl ow rate of 0.4 ml/min. A UPLC run time of 20 min was routinely used, and a shorter method with a 10 min run time is also described. The method shows extremely stable retention times when human plasma extracts and a variety of biofl uids or tissues are analyzed [intra-assay relative standard deviation (RSD) <0.385% and <0.451% for 20 and 10 min gradients, respectively (n = 5); interassay RSD <0.673% and <0.763% for 20 and 10 min gradients, respectively (n = 30)]. The UPLC system was coupled to a hybrid quadrupole orthogonal acceleration time-of-fl ight mass spectrometer, equipped with a traveling wave ion-mobility cell. Besides demonstrating the separation for different lipids using the chromatographic method, we demonstrate the use of the ion-mobility MS platform for the structural elucidation of lipids. The method can now be used to elucidate structures of a wide variety of lipid...

show abstract

“…Nevertheless, these methods were hampered either by limited throughput in using SPE, or by inconstant factors of dripping loss or possible interfering protein in using chloroform/methanol/water. Currently MTBE extraction was used as a substitute for chloroform extraction [26,34]. Hence we proposed a procedure using MTBE combined with mild alkaline hydrolysis as described in Section 2.3.…”

Section: Serum Sphingolipid Extractionmentioning

confidence: 99%

Large-scaled human serum sphingolipid profiling by using reversed-phase liquid chromatography coupled with dynamic multiple reaction monitoring of mass spectrometry: Method development and application in hepatocellular carcinoma

Simultaneous extraction of metabolome and lipidome with methyl tert-butyl ether from a single small tissue sample for ultra-high performance liquid chromatography/mass spectrometry

Cited by 190 publications

References 38 publications

Integration of Metabolomics and Transcriptomics Reveals Major Metabolic Pathways and Potential Biomarker Involved in Prostate Cancer

Integration of Metabolomics and Transcriptomics Reveals Major Metabolic Pathways and Potential Biomarker Involved in Prostate Cancer

Enhanced lipid isomer separation in human plasma using reversed-phase UPLC with ion-mobility/high-resolution MS detection

Large-scaled human serum sphingolipid profiling by using reversed-phase liquid chromatography coupled with dynamic multiple reaction monitoring of mass spectrometry: Method development and application in hepatocellular carcinoma

Contact Info

Product

Resources

About