" How do I take the dilutions from above and make a “spot chart” I can compare my experimental data to?"
To create a spot chart for cannabinoid quantitation using TLC, you will need to:
- Prepare a series of different concentrations of the cannabinoid standard solution.
- Apply each concentration of the cannabinoid standard solution to the TLC plate, starting with the lowest concentration and working your way up to the highest concentration.
- Develop the TLC plate and visualize the cannabinoid spots as described above.
- Mark the location of each cannabinoid spot on the TLC plate.
- Label each cannabinoid spot with the concentration of the cannabinoid standard solution that was applied to the TLC plate.
The resulting spot chart will serve as a reference for comparing the cannabinoid spots in your experimental sample.
Another way to create a spot chart is to use a TLC scanner. A TLC scanner is a device that can scan the TLC plate and create a digital image of the cannabinoid spots. The digital image can then be used to create a spot chart.
To create a spot chart using a TLC scanner, you will need to:
- Scan the TLC plate using the TLC scanner.
- Save the digital image of the TLC plate to your computer.
- Use a software program to analyze the digital image and identify the cannabinoid spots.
- The software program will typically generate a report that includes the location and intensity of each cannabinoid spot.
- You can use the report to create a spot chart by plotting the location of each cannabinoid spot against the intensity of the spot.
Once you have created a spot chart, you can use it to compare the cannabinoid spots in your experimental sample to the cannabinoid spots in the standard curve. This will allow you to identify the cannabinoids in your sample and to quantitate the amount of each cannabinoid.
Here are some tips for creating a spot chart:
- Use a consistent mobile phase and developing time when creating the spot chart. This will ensure that the cannabinoid spots are reproducible.
- Apply a wide range of concentrations of the cannabinoid standard solution to the TLC plate. This will create a more accurate spot chart.
- Use a densitometer or TLC scanner to measure the intensity of the cannabinoid spots. This will provide you with more accurate results.
- Label the cannabinoid spots on the spot chart with the concentration of the cannabinoid standard solution that was applied to the TLC plate. This will make it easier to compare the cannabinoid spots in your experimental sample to the cannabinoid spots in the standard curve.
…and while the human eye is a incredible sensor, I recommend the electronic method rather than spotting, developing, “photocopying” to transparent media any day.
Edit: FYI, the AI has come pretty close, but really doesn’t understand quite what you’re after. You should however be able to leverage it.
Check @Lincoln20XX’s posts on hplc method dev to get a better understanding of why you need calibrate your measuring devices.