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Pharmacokinetics

Comprehensive Metabolite Analysis GCMS-TQ8040 NX

LCMS-8040 / 8050

Comprehensive Analysis of All Metabolites Using

GC/MS and LC/MS for Researching Intestinal Bacteria Cell Line Optimization
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Operating Principle and Features High Peptides LC/MS • The world’s largest metabolite database includes preregistered optimized methods.
Coenzymes,
Metabolomic analysis using a mass spectrometer generally involves using a Molecular weight Nucleotides, Lipids • MRM measurements can detect components not detectable by scan or SIM modes.
Steroids, Vitamins
gas chromatograph mass spectrometer (GC-MS) or high-performance liquid Nucleosides, • High-speed MRM analysis enables simultaneous analysis of multiple components to
Terpenes
Sugar phosphates
chromatograph mass spectrometer (LC-MS) to comprehensively analyze Hydrocarbons Sugars, Amino acids, Culture
all the metabolites (metabolome) contained in a sample. That requires Esters Organic acids achieve comprehensive high-sensitivity analysis.
selectively using GC/MS or LC/MS based on the target components being Ketones Fatty acids
Alcohol
analyzed or the given purpose of analysis, as illustrated in Fig. 1. Using a
GC/MS to analyze hydrophilic metabolites such as amino acids, organic Low GC/MS Non-volatile
Volatile
acids, or sugars requires a derivatization process, but it offers superior Fig. 1 Target Components for GC/MS and LC/MS Analysis
robustness and can comprehensively analyze hundreds of components
in a single analysis. In contrast, an LC/MS can efficiently analyze specific GC/MS/MS
metabolites (up to 100 components) without derivatization, making it well- 750000 (Smart Metabolites Database)
suited for routine analysis of specific components. 500000
250000 Purification
Measurement Method and Conditions 0
(x100,000) 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 16.0 17.0 18.0 19.0
6.0
4.0
Fresh fecal samples were collected from male C57BL/6J mice raised in a 3.0 (LC/MS/MS Method Package for Primary Metabolites Ver. 2)
Ion pair LC/MS/MS
normal environment. 450 μL of a physiological phosphate buffer solution 2.0
was added to 50 mg of the fecal samples and then stirred. Then the 1.0
supernatant was ultrafiltered by centrifugal separation to extract the 0.0
0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 min
metabolites. To analyze the primary metabolites by GC-MS/MS, the filtrate (x10,000,000)
Ion pair-free LC/MS/MS
was derivatized to prepare the samples for GC-MS/MS. Meanwhile, to 1.5 (LC/MS/MS Method Package for Primary Metabolites Ver. 2)
analyze the primary metabolites by LC-MS/MS, the filtrate was diluted by 1.0
ten times with ultrapure water in preparation for LC-MS/MS. 0.5
For the GC/MS/MS analysis, 475 components were analyzed simultaneously 0.0 Characterization
0.0 2.5 5.0 7.5 10.0 12.5 15.0 min
using an MRM method from the Smart Metabolites Database, which Fig. 2 MRM Chromatograms of Mouse Fecal Extract
includes MRM information for 475 components, mainly for metabolites
included in biological samples. For the LC/MS/MS analysis, a method of
ion pair LC/MS/MS and a method of ion pair-free LC/MS/MS were used
for analysis in LCMS-8040 and LCMS-8050 systems. The method of ion
pair LC/MS/MS is intended for simultaneous analysis of 55 metabolite
components important for metabolomic analysis in the life sciences, such
as for analyzing the glycolytic system, TCA cycle, pentose phosphate
pathway, or amino acids/nucleotides, whereas the method of ion pair-free Quality Control
LC/MS/MS is intended for simultaneous analysis of 97 organic acid and
other metabolite components that cannot be analyzed using the method
of ion pair LC/MS/MS. Both methods are included in the LC/MS/MS
Method Package for Primary Metabolites Ver. 2. Fig. 3 Number of Metabolites Detected from Mouse Fecal Extract
Conclusion
Results
Because GC/MS/MS and LC/MS/MS methods target different Specifications
The GC/MS/MS analysis detected 100 components, mostly short-chain components, comprehensive analysis of metabolites is enabled by
fatty acids and organic acids. It even detected 17 sugar components using both methods. Furthermore, by using the MRM database for Instrument GCMS-TQ8040 NX
that are difficult to analyze by LC or LC/MS/MS (Fig. 2). The ion pair GC/MS/MS analysis and method packages for LC/MS/MS analysis, Pharmacokinetics
method detected 17 components, including mainly amino acids. The comprehensive analysis can be easily performed by operators who GC unit Oven temperature: Room temperature + 2 to 450 °C
ion pair-free method detected 75 components, including amino acids, are not very familiar with this analysis. The large amounts of data Carrier gas control: Constant liner velocity, constant pressure, or constant flowrate
Flow controller pressure: Max. 970 kPa
nucleotides, nucleosides, and organic acids involved in the TCA cycle (Fig. 2). generated can be interpreted easily by using Shimadzu’s Multi-Omic
Therefore, it is extremely useful to use both GC/MS/MS and LC/MS/MS for Data Analysis package to visualize the data. These sample and data MS unit Ionization: EI (standard), CI, NCI (optional)
comprehensively analyzing metabolites in fecal samples. analysis methods should be extremely useful not only for researching Mass range: m/z 10 to 1090
intestinal flora, but also for metabolomic analysis in a wide variety of MRM max. speed: >800 MRM/sec
This article was prepared with generous cooperation from Takanari Hattori *1, *2 , Akihiko other pharmacokinetic applications. Measurement modes: Scan mode, SIM mode, MRM mode,
*1
*1
Kunisawa *1,*2 , Shuichi Kawano , Shinichi Kono *1,*2 , Yoshihiro Hayakawa , Junko Iida *1,*2 , Eiichiro product ion scan mode, Others
*4
Fukusaki *2,*3 , Mitsuharu Matsumoto . precursor ion scan mode,
*1 Analytical & Measuring Instruments Division, Shimadzu Corporation Application Examples and neutral loss scan mode,
*2 Osaka University and Shimadzu Analytical Innovation Research Laboratory or simultaneous analysis with any combination thereof.
*3 Graduate School of Engineering, Osaka University
*4 Research Laboratories, Kyodo Milk Industry Co., Ltd. • Simultaneous analysis of metabolites (metabolomics) Weight: 110 kg for GC-MS main units and 10 kg for auxiliary pump
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