TY - JOUR
T1 - Quantitative metabolic network profiling of Escherichia coli
T2 - An overview of analytical methods for measurement of intracellular metabolites
AU - Shen, Yang
AU - Fatemeh, Tohidi
AU - TANG, Lei Han
AU - Cai, Zongwei
N1 - Funding Information:
The authors are benefitted from the discussion with Zhi Tang and Xiaona Li. Financial supports from Collaborative Research Fund of Hong Kong Research Grant Council ( HKBU5/CRF/10 ) and Hong Kong Baptist University ( IRMC/12-13/1B ) are acknowledged.
PY - 2016/1
Y1 - 2016/1
N2 - Understanding the mechanism of metabolic network response to extracellular changes has become one of the primary objectives of systems biology. Quantitative metabolic network profiling of model organisms such as E. coli is the principal approach. Well-designed analytical protocols for the determination of intracellular metabolites may provide a crucial premise to quantitative profiling of cellular metabolism. This article reviews and evaluates the existing methods for determining intracellular metabolites of E. coli in terms of sample preparation and mass spectrometry (MS) analysis. Critical issues including the variation of metabolic status and interference of external pools during cell sampling, sample loss during quenching procedures, degradation of metabolites and efficiency of extraction, and matrix effects in separation and quantification are discussed. Moreover, suggestions are provided to solve the analytical problems based on the results from the recent studies in order to establish a precise and reliable analytical platform for the intracellular metabolites.
AB - Understanding the mechanism of metabolic network response to extracellular changes has become one of the primary objectives of systems biology. Quantitative metabolic network profiling of model organisms such as E. coli is the principal approach. Well-designed analytical protocols for the determination of intracellular metabolites may provide a crucial premise to quantitative profiling of cellular metabolism. This article reviews and evaluates the existing methods for determining intracellular metabolites of E. coli in terms of sample preparation and mass spectrometry (MS) analysis. Critical issues including the variation of metabolic status and interference of external pools during cell sampling, sample loss during quenching procedures, degradation of metabolites and efficiency of extraction, and matrix effects in separation and quantification are discussed. Moreover, suggestions are provided to solve the analytical problems based on the results from the recent studies in order to establish a precise and reliable analytical platform for the intracellular metabolites.
KW - Escherichia coli
KW - Intracellular metabolites
KW - Mass spectrometry
KW - Metabolic network profiling
KW - Sample preparation
UR - http://www.scopus.com/inward/record.url?scp=84947976549&partnerID=8YFLogxK
U2 - 10.1016/j.trac.2015.07.006
DO - 10.1016/j.trac.2015.07.006
M3 - Review article
AN - SCOPUS:84947976549
SN - 0165-9936
VL - 75
SP - 141
EP - 150
JO - TrAC - Trends in Analytical Chemistry
JF - TrAC - Trends in Analytical Chemistry
ER -