Dear All. I have separated 38 amino acids on HILIC column couple to QQQ followed by using 10 isotope-labeled amino acids as internal standard. However, I am not sure which formula to use to calculate the concentration of my amino acids. Can you help me out? There several formal out in literature I not sure which is one is best????
Thanks ...
For each AA, you need to create and run a full calibration that spans the range of concentrations tested (at least 5 - points per order of magnitude). Once you have the calibration curves created and saved for each standard and incorporated into the analysis method, you can then analyze each sample, using bracketed standards as needed and your IS to account for any sample prep or other matrix corrections.
What are bracketed standards? Can you explain that in the formula? I have a lot of samples I want to write a program for it. Thanks... Do you have any literature on Which I can look at the formula?
- "Bracketed standards" = This is the statistical process of running a set of standards, followed by one or more samples, followed by another set of standards. This is done to compensate for any 'changes' that may occur over time. The standards 'bracket' the samples (before/after) allowing for averaging of and/or weighting of any changes noted over time. We use these types of techniques to account for variations which occur when we run large numbers of samples.
What you need to do is to speak with a local, experienced chromatographer (way too much to go into on a forum like this! It takes many years of training to learn HPLC method development and someone with the training could help you set all of this up. No programming needed). Any experienced HPLC chromatographer will know all about how to set-up, create and run a quantitative analysis using standards, with or without bracketing, that is appropriate for your application. All of the professional chromatography software packages have these features built-in to them as they are used all of the time. Good luck to you.
Article Accurate and efficient amino acid analysis for protein quant...
- Badges
- Science topic
- Similar topics
- Analytical Chemistry
- Column