Method Package for Cell Culture Profiling
Описание
For LabSolutions LCMS
Simultaneous Analysis Conditions for 125 Compounds
This Method Package enables the analysis of 125 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.
Compared to the previous version, several metabolites derived from amino acids, nucleic acids and the TCA cycle have been added, allowing even more comprehensive profiling information to be gathered.
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.
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 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 Citrulline Ornithine Proline Putrescine Argininosuccinic acid 2-Aminobutyric acid 5-Glutamylcysteine 5′-Methylthioadenosine Cystathionine Cysteine Cystine Glutathione Homocysteine Methionine Methionine sulfoxide N-Acetylcysteine Oxidized glutathione S-Adenosylhomocysteine Glycine Serine Threonine 1-Methylhistidine 3-Methylhistidine Histidine Urocanic acid 2-Aminoadipic acid Hydroxylysine Lysine Pipecolic acid Saccharopine 3-Hydroxyanthranilic acid 5-Hydroxytryptophan Anthranilic acid Formylkynurenine Hydroxykynurenine Indole-3-acetic acid Kynurenic acid Kynurenine
Serotonin Tryptophan 4-Hydroxyphenyllactic acid Phenylalanine Tyrosine 3-Hydroxyisobutyric acid 3-Methyl-2-oxovaleric acid Isoleucine Leucine Valine Alanyl-glutamine Glycyl-glutamine |
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 MT Arg MT Arg MT Arg MT Arg MT Arg MT Arg MT Arg 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 His MT His MT His MT His 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 Tyr MT Tyr MT Tyr MT Val MT Val MT Val MT Val MT Val MT — — |
| Others | Group |
|---|---|
| 2-ketoglutaric acid Acotinic acid Citric acid Fumaric acid Isocitric acid Lactic acid Malic acid Pyruvic acid Succinic acid Penicillin G 2-Aminoethanol Glyceric acid NAD O-Phosphoethanolamine Taurine |
TCA Cycle TCA Cycle TCA Cycle TCA Cycle TCA Cycle TCA Cycle TCA Cycle TCA Cycle TCA Cycle Antibiotics — — — — — |
| 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 Niacinamide Nicotinic acid Pantothenic acid 4-Pyridoxic acid Pyridoxal Pyridoxalphosphate Pyridoxine Biotin 4-Aminobenzoic acid Folic acid Choline Ascorbic acid Cyanocobalamin Lipoic acid |
B2 B3 B3 B5 B6 B6 B6 B6 B7 B9 B9 B12 C Vitamin-like Vitamin-like |
| Internal standard | Group |
|---|---|
| 2-Isopropylmalic acid |
Remarks and Precautions
LabSolutions LCMS Ver.5.97 or later is required.
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.
LabSolutions and LCMS are trademarks of Shimadzu Corporation.
GARUDA is a trademark of The Systems Biology Institute.
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Applications
Simultaneous Analysis of Culture Supernatant of Mammalian Cells Using Triple Quadrupole LC/MS/MS
Industrial fermentation for the production of biofuels or biopharmaceutics requires routine monitoring of medium conditions such as pH, dissolved gas, carbon source (glucose) and nitrogen source (glutamine) for optimization and control of the fermentation process. However, culture media also consist of various other biologically important compounds such as vitamins, nucleic acids and other primary metabolites, which would lead to more detailed understanding of the bioprocess if monitored altogether.
*The data was obtained by LC/MS/MS Method Package for Cell Culture Profiling Ver.1.
