LC/MS/MS Method Package for Cell Culture Profiling Ver. 3

For LabSolutions™ LCMS Software

Simultaneous Analysis Conditions for 144 Compounds

This Method Package enables the analysis of 144 compounds in under 20 minutes per sample. Performing analysis separately for each compound group such as amino acids and vitamins makes profiling of cell culture components very laborious, but with this method package a large number of culture medium components and secreted metabolites can be analyzed simultaneously.

Version 3 of this method package adds metabolites characteristic of CHO cells, which are frequently used in the manufacture of monoclonal antibodies



 

Optimized Method for Culture Medium Analysis

Pre-set analysis conditions make full use of triple quadrupole MS capabilities for analyzing trace components such as vitamins. On the other hand, analysis conditions are adjusted so that high-concentration components such as glucose or amino acids are not saturated. It is possible to measure a variety of culture medium components from the same vial.

Note: A dilution series must be created for accurate quantification.

 


Analysis of culture supernatant 

 

Ready-to-Use Method

Analysis can be started without the need for time-consuming preparations such as optimization of MS parameters for each compound or careful consideration of LC/MS/MS analysis conditions.


Excellent Data Analysis Functionality

Using the Multiomics Analysis Package, it is easy and straightforward to compare changes in compound quantities on a bar graph, display changes over time on a line chart, etc.

It also includes easy-to-implement functions for correlation analysis, volcano plots, and more, significantly reducing the time required for bottleneck processes such as data analysis and visualization. The whole workflow from measurement to data analysis becomes smoother and more efficient.

 



 

Data obtained with the method package can be inserted on the included metabolic map. It is then simple to compare compound quantities on a bar graph or display changes over time on a line graph.



 

The data analysis software in this Method Package is developed based on tools (gadgets) that have been released on the GARUDA™ open research platform, which is mainly managed by The Systems Biology Institute, Japan (SBI).



 

Data Analysis Tools Used in Method Package

Volcano Plot
A tool that combines a t-test (statistically significant difference) and a fold analysis (Example: Difference in mean value such as 2 times or 1/2) to visualize the differences between the 2 groups. The Volcano Plot gadget developed by Shimadzu is included in the package.

VANTED
Tool maintained at University of Konstanz, Germany, for visualization and analysis of networks across different data sets. (GARUDA support was developed at Monash University)

iPath
Data analysis tool developed by the European Molecular Biology Laboratory that can be used for visualization of diverse metabolic pathways or data mapping and customization.

Cytoscape
Bioinformatics tool developed by the Cytoscape Consortium, used to visualize metabolic pathways, to integrate gene expression profiles with related data, and so on. It is especially useful for analyzing networks and visualizing correlations.

List of Registered Compounds

 
Amino acids and their metabolites Group
5-Oxoproline
Alanine
Asparagine
Aspartic acid
Glutamic acid
Glutamine
N-Acetylaspartic acid
4-Aminobutyric acid
4-Hydroxyproline
Arginine
Argininosuccinic acid
Citrulline
Creatine
Ornithine
Proline
Putrescine
2-Aminobutyric acid
5-Glutamylcysteine※
5'-Methylthioadenosine
Cystathionine
Cysteine
Cystine
Glutathione
Homocysteine
Homocystine
Methionine
Methionine sulfoxide
N-Acetylcysteine
Ophthalmic acid
Oxidized glutathione
S-Adenosylhomocysteine
2-Aminoethanol
Glycine
O-Phosphoethanolamine
Serine
Threonine
2-Aminoadipic acid
Acetylcarnitine
Carnitine
Hydroxylysine
Lysine
Pipecolic acid
Saccharopine
3-Hydroxyanthranilic acid
5-Hydroxytryptophan
Anthranilic acid
Formylkynurenine
Hydroxykynurenine
Indole-3-acetic acid
Kynurenic acid
Kynurenine
Serotonin
Tryptophan
Ala, Asp and Glu MT
Ala, Asp and Glu MT
Ala, Asp and Glu MT
Ala, Asp and Glu MT
Ala, Asp and Glu MT
Ala, Asp and Glu MT
Ala, Asp and Glu MT
Arg and Pro MT
Arg and Pro MT
Arg and Pro MT
Arg and Pro MT
Arg and Pro MT
Arg and Pro MT
Arg and Pro MT
Arg and Pro MT
Arg and Pro MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Cys and Met MT
Gly and Ser MT
Gly and Ser MT
Gly and Ser MT
Gly and Ser MT
Gly and Ser MT
Lys MT
Lys MT
Lys MT
Lys MT
Lys MT
Lys MT
Lys MT
Trp MT
Trp MT
Trp MT
Trp MT
Trp MT
Trp MT
Trp MT
Trp MT
Trp MT
Trp MT
Amino acids and their metabolites Group
1-Methylhistidine
3-Methylhistidine
Histidine
Urocanic acid
4-Hydroxyphenyllactic acid
Dopa
Norepinephrine
Phenylalanine
Phenyllactic acid
Tyrosine
3-Hydroxyisobutyric acid
3-Methyl-2-oxovaleric acid
Isoleucine
Leucine
Valine
Alanyl-glutamine
Asymmetric dimetyhlarginine
Glycyl-glutamine
Symmetric dimetyhlarginine
His MT
His MT
His MT
His MT
Tyr and Phe MT
Tyr and Phe MT
Tyr and Phe MT
Tyr and Phe MT
Tyr and Phe MT
Tyr and Phe MT
Val MT
Val MT
Val MT
Val MT
Val MT
Others
Others
Others
Others
Others Group
2-ketoglutaric acid
Acotinic acid
Citric acid
Fumaric acid
Isocitric acid
Lactic acid
Malic acid
Pyruvic acid
Succinic acid
Glyceric acid
Glycolic acid
Glyoxylic acid
Mevalonic acid
MVA-P
Penicillin G
Resveratrol
Shikimic acid 3-phosphate
Taurine
TCA Cycle
TCA Cycle
TCA Cycle
TCA Cycle
TCA Cycle
TCA Cycle
TCA Cycle
TCA Cycle
TCA Cycle
Others
Others
Others
Others
Others
Others
Others
Others
Others
 
Nucleic acids and their metabolites Group
Adenine
Adenosine
Adenosine monophosphate
Deoxyadenosine
Deoxyadenosine monophosphate
Deoxyguanosine
Deoxyguanosine monophosphate
Guanine
Guanosine
Guanosine monophosphate
Hypoxanthine
Inosine
Inosine monophosphate
Uric acid
Xanthine
Xanthosine
Xanthosine monophosphate
3-Aminoisobutyric acid
3-Aminopropanoic acid
Cytidine
Cytidine monophosphate
Cytosine
Deoxycytidine
Deoxycytidine monophosphate
Orotic acid
Thymidine
Thymidine monophosphate
Thymine
Uracil
Uridine
Uridine monophosphate
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Purine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Pyrimidine MT
Sugars Group
Gluconic acid                    
Hexose (Glucose)
Sucrose
Threonic acid



Vitamins Group
Riboflavin
Pantothenic acid
4-Pyridoxic acid
Pyridoxal
Pyridoxalphosphate
Pyridoxine
Biotin
4-Aminobenzoic acid
Folic acid
Cyanocobalamin
Ascorbic acid
Acetylcholine
Choline
Citicoline
NAD
Niacinamide
Nicotinic acid
Lipoic acid
B2
B6
B6
B6
B6
B7
B9
B9
B12
C
Others
Others
Others
Others
Others
Others
Others
Internal standard Group
2-Isopropylmalic acid
10-Camphorsulfonic acid

※This compound may be deleted from the list depending on the analytical column selected.
 

Remarks and Precautions

  1. LabSolutions LCMS Ver.5.113 or later is required, and LabSolutiosn Insight Ver. 3.8 SP3 or later is required.
  2. It is the user’s responsibility to adopt appropriate quality control tests using standard samples to confirm qualitative and quantitative information obtained with this method package.
  3. This method package is intended for research use. It may not be used for clinical diagnostic applicaitons.
 

LabSolutions and LCMS are trademarks of Shimadzu Corporation.
GARUDA is a trademark of The Systems Biology Institute.

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