An optimized procedure for metabonomic analysis of rat liver tissue using gas chromatography/time-of-flight mass spectrometry

Li Pan, Yunping Qiu, Tianlu Chen, Jinchao Lin, Yi Chi, Mingming Su, Aihua Zhao, Wei Jia

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

In this paper, we present a tissue metabonomic method with an optimized extraction procedure followed by instrumental analysis with gas chromatography/time-of-flight mass spectrometry (GC/TOFMS) and spectral data analysis with multivariate statistics. Metabolite extractions were carried out using three solvents: chloroform, methanol, and water, with design of experiment (DOE) theory and multivariate statistical analysis. A two-step metabolite extraction procedure was optimized using a mixed solvent of chloroform-methanol-water (1:2:1, v/v/v) and then followed by methanol alone. This approach was subsequently validated using standard compounds and liver tissues. Calibration curves were obtained in the range of 0.50-125.0μg/mL for standards and 0.02-0.25 g/mL acceptable for liver tissue samples. For most of the metabolites investigated, relative standard deviations (RSD) were below 10% within a day (reproducibility) and below 15% within a week (stability). Rat liver tissues of carbon tetrachloride-induced acute liver injury models (n=. 10) and healthy control rats (n=. 10) were analyzed which demonstrated the applicability of the developed procedure for the tissue metabonomic study.

Original languageEnglish (US)
Pages (from-to)589-596
Number of pages8
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume52
Issue number4
DOIs
StatePublished - Aug 2010
Externally publishedYes

Fingerprint

Metabolomics
Gas chromatography
Gas Chromatography
Liver
Mass spectrometry
Rats
Mass Spectrometry
Tissue
Metabolites
Methanol
Chloroform
Rat control
Water
Carbon Tetrachloride
Design of experiments
Calibration
Statistical methods
Multivariate Analysis
Statistics
Wounds and Injuries

Keywords

  • Carbon tetrachloride
  • Gas chromatography/time-of-flight mass spectrometry
  • Liver tissue
  • Metabonomics
  • Multivariate statistics

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmaceutical Science
  • Spectroscopy
  • Clinical Biochemistry
  • Medicine(all)

Cite this

An optimized procedure for metabonomic analysis of rat liver tissue using gas chromatography/time-of-flight mass spectrometry. / Pan, Li; Qiu, Yunping; Chen, Tianlu; Lin, Jinchao; Chi, Yi; Su, Mingming; Zhao, Aihua; Jia, Wei.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 52, No. 4, 08.2010, p. 589-596.

Research output: Contribution to journalArticle

Pan, Li ; Qiu, Yunping ; Chen, Tianlu ; Lin, Jinchao ; Chi, Yi ; Su, Mingming ; Zhao, Aihua ; Jia, Wei. / An optimized procedure for metabonomic analysis of rat liver tissue using gas chromatography/time-of-flight mass spectrometry. In: Journal of Pharmaceutical and Biomedical Analysis. 2010 ; Vol. 52, No. 4. pp. 589-596.
@article{22f4c5b37a914e64b1dd753be86ae1f9,
title = "An optimized procedure for metabonomic analysis of rat liver tissue using gas chromatography/time-of-flight mass spectrometry",
abstract = "In this paper, we present a tissue metabonomic method with an optimized extraction procedure followed by instrumental analysis with gas chromatography/time-of-flight mass spectrometry (GC/TOFMS) and spectral data analysis with multivariate statistics. Metabolite extractions were carried out using three solvents: chloroform, methanol, and water, with design of experiment (DOE) theory and multivariate statistical analysis. A two-step metabolite extraction procedure was optimized using a mixed solvent of chloroform-methanol-water (1:2:1, v/v/v) and then followed by methanol alone. This approach was subsequently validated using standard compounds and liver tissues. Calibration curves were obtained in the range of 0.50-125.0μg/mL for standards and 0.02-0.25 g/mL acceptable for liver tissue samples. For most of the metabolites investigated, relative standard deviations (RSD) were below 10{\%} within a day (reproducibility) and below 15{\%} within a week (stability). Rat liver tissues of carbon tetrachloride-induced acute liver injury models (n=. 10) and healthy control rats (n=. 10) were analyzed which demonstrated the applicability of the developed procedure for the tissue metabonomic study.",
keywords = "Carbon tetrachloride, Gas chromatography/time-of-flight mass spectrometry, Liver tissue, Metabonomics, Multivariate statistics",
author = "Li Pan and Yunping Qiu and Tianlu Chen and Jinchao Lin and Yi Chi and Mingming Su and Aihua Zhao and Wei Jia",
year = "2010",
month = "8",
doi = "10.1016/j.jpba.2010.01.046",
language = "English (US)",
volume = "52",
pages = "589--596",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
issn = "0731-7085",
publisher = "Elsevier",
number = "4",

}

TY - JOUR

T1 - An optimized procedure for metabonomic analysis of rat liver tissue using gas chromatography/time-of-flight mass spectrometry

AU - Pan, Li

AU - Qiu, Yunping

AU - Chen, Tianlu

AU - Lin, Jinchao

AU - Chi, Yi

AU - Su, Mingming

AU - Zhao, Aihua

AU - Jia, Wei

PY - 2010/8

Y1 - 2010/8

N2 - In this paper, we present a tissue metabonomic method with an optimized extraction procedure followed by instrumental analysis with gas chromatography/time-of-flight mass spectrometry (GC/TOFMS) and spectral data analysis with multivariate statistics. Metabolite extractions were carried out using three solvents: chloroform, methanol, and water, with design of experiment (DOE) theory and multivariate statistical analysis. A two-step metabolite extraction procedure was optimized using a mixed solvent of chloroform-methanol-water (1:2:1, v/v/v) and then followed by methanol alone. This approach was subsequently validated using standard compounds and liver tissues. Calibration curves were obtained in the range of 0.50-125.0μg/mL for standards and 0.02-0.25 g/mL acceptable for liver tissue samples. For most of the metabolites investigated, relative standard deviations (RSD) were below 10% within a day (reproducibility) and below 15% within a week (stability). Rat liver tissues of carbon tetrachloride-induced acute liver injury models (n=. 10) and healthy control rats (n=. 10) were analyzed which demonstrated the applicability of the developed procedure for the tissue metabonomic study.

AB - In this paper, we present a tissue metabonomic method with an optimized extraction procedure followed by instrumental analysis with gas chromatography/time-of-flight mass spectrometry (GC/TOFMS) and spectral data analysis with multivariate statistics. Metabolite extractions were carried out using three solvents: chloroform, methanol, and water, with design of experiment (DOE) theory and multivariate statistical analysis. A two-step metabolite extraction procedure was optimized using a mixed solvent of chloroform-methanol-water (1:2:1, v/v/v) and then followed by methanol alone. This approach was subsequently validated using standard compounds and liver tissues. Calibration curves were obtained in the range of 0.50-125.0μg/mL for standards and 0.02-0.25 g/mL acceptable for liver tissue samples. For most of the metabolites investigated, relative standard deviations (RSD) were below 10% within a day (reproducibility) and below 15% within a week (stability). Rat liver tissues of carbon tetrachloride-induced acute liver injury models (n=. 10) and healthy control rats (n=. 10) were analyzed which demonstrated the applicability of the developed procedure for the tissue metabonomic study.

KW - Carbon tetrachloride

KW - Gas chromatography/time-of-flight mass spectrometry

KW - Liver tissue

KW - Metabonomics

KW - Multivariate statistics

UR - http://www.scopus.com/inward/record.url?scp=77950369068&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77950369068&partnerID=8YFLogxK

U2 - 10.1016/j.jpba.2010.01.046

DO - 10.1016/j.jpba.2010.01.046

M3 - Article

C2 - 20185264

AN - SCOPUS:77950369068

VL - 52

SP - 589

EP - 596

JO - Journal of Pharmaceutical and Biomedical Analysis

JF - Journal of Pharmaceutical and Biomedical Analysis

SN - 0731-7085

IS - 4

ER -