How much internal standard is added in targeted metabolomics?

The amount of internal standard added in targeted metabolomics analysis depends on various factors, including sample type, analytical method, instrument sensitivity, and the concentration range of target metabolites. Here are some general guidelines for your reference:

1. Selection of an Appropriate Internal Standard

• Isotopically labeled internal standards: Use isotopically labeled internal standards that have a similar structure to the target metabolites but with different masses.
• Structurally similar internal standards: If isotopically labeled internal standards are not available, choose compounds with a similar structure as the internal standard.

2. Determining the Amount of Internal Standard to Add

• Standard curve method: Prepare a standard solution of the internal standard within a known concentration range and add it to the sample, ensuring that the internal standard concentration is within the linear range of the analytical method.
• Empirical values: Choose an appropriate amount of internal standard based on past experience and literature. Generally, the internal standard concentration should be similar to the concentration of the target metabolites to ensure accurate quantitative analysis.

3. Specific Steps

• Sample preparation: Add the internal standard at an early stage of sample processing to compensate for any losses and changes during the sample processing and analysis.
• Concentration range: The concentration of the internal standard should be within the expected concentration range of the target metabolites, typically 10-100 times the concentration of the target metabolites, to ensure reliable quantitative results.

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