Separation / Chromatography
Lets assume I have three component mixture of liquids. And I wish to determine the quantity of each component present. I have reference samples of two of the components but the other component is not identified. How do I design an experiment in order to obtain the % composition of the mixture.
I don't expect you to write whole experiment, just give me some start off. I don't have any idea to start this experiment design.
Thank you!
See my response above.
To determine the composition of the three-component mixture, you can design an experiment using chromatography. Chromatography is a technique that separates components of a mixture based on their different properties and allows for their identification and quantification. Here are the steps you can follow to get started with your experiment design:
1. Choose a suitable chromatography technique: There are various chromatographic techniques available, such as paper chromatography, thin-layer chromatography (TLC), gas chromatography (GC), or high-performance liquid chromatography (HPLC). You need to select the appropriate technique based on the nature of your mixture and the components you want to separate.
2. Set up the chromatography system: The setup depends on the chosen technique. For example, in paper chromatography, you will need a strip of filter paper, a developing solvent, and a container to hold the solvent. In TLC, a thin layer of adsorbent material on a plate is needed, along with a developing chamber. For GC or HPLC, specialized equipment and columns are required.
3. Prepare the mixture: Take a known volume of the three-component mixture and dilute it if necessary. You should have reference samples of the known components for comparison.
4. Apply the sample: In the chosen chromatography technique, apply a small amount of the mixture and the reference samples onto the appropriate medium (e.g., filter paper, TLC plate, or column).
5. Develop the chromatogram: Place the medium into the container with the developing solvent. The solvent will move up the medium, carrying the components with them. Each component will have a different affinity for the medium and solvent, causing them to separate as distinct spots or bands on the medium.
6. Visualize the separated components: Once the solvent reaches the desired height on the medium, remove it, and let it dry. Depending on the technique, you may use UV light, staining reagents, or other detection methods to visualize the separated components.
7. Measure the distances traveled: Measure the distances each component and the reference samples have traveled from the application point to the solvent front. Typically, you measure the distance in chromatography in terms of "Rf" value, which is the ratio of the distance traveled by the component to the distance traveled by the solvent front.
8. Compare the Rf values: Compare the Rf values of the separated components in the mixture with the Rf values of the reference samples. If the unknown component has a similar Rf value to one of the reference samples, you can assign it accordingly. If not, it may indicate an unidentified component.
9. Calculate the % composition: If you have identified all three components, you can calculate their relative percentages in the mixture by comparing their Rf values. This calculation can be done using the ratio of the distances traveled or by comparing the intensities of the spots or bands observed.
Remember, the success of the experiment depends on choosing the right chromatography technique, optimized conditions, and proper analysis of the results obtained. It is also important to have appropriate reference samples and accurate measurements for reliable quantification.