Chiral Purity (enantiomer excess) for HPLC

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In summary, the conversation discusses how to calculate Chiral Purity for HPLC testing using peak area. The ratio of the areas of both enantiomers is used to determine the enantiomeric excess (ee), which is then converted into a percentage. The desired enantiomer can be determined by the sign of the resulting number, with a negative number indicating the production of the undesired product.
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cwj7316
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[SOLVED] Chiral Purity (enantiomer excess) for HPLC

Hello!

When I use HPLC to test Chiral Purity, obtain chromatogra and peak area. How do I calculate with Peak area?

thanks!
 
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  • #2
cwj7316 said:
Hello!

When I use HPLC to test Chiral Purity, obtain chromatogra and peak area. How do I calculate with Peak area?

thanks!

LOL you want to know how they used to do it back in the day? Weigh the piece of paper your spectrum is on. Cut out the peaks and weigh the paper. You should be able to tell by the difference in mass how pure your compound is.
 
  • #3
If you are given the areas of both enantiomers (or even epimers), the ee is calculated by first summing the areas of both peaks and determining their relative ratios. R and S, for example where R+S=1 and ratio of R=R/(R+S). ee is defined as {(R-S)/(R+S)}*100. If the areas are 0.6:0.4, the ee is 20%. If the areas are 0.98:0.02, the ee would be 0.96. In this example the desired enantiomer is R. If it is S in your case, just switch it around. If you get a negative number, the reaction produces an ee of the undesired product.
 
  • #4
Thanks gravenewworld and chemisttree.
 

1. What is chiral purity and why is it important for HPLC?

Chiral purity, also known as enantiomer excess, is a measure of the percentage of a single enantiomer present in a sample. It is important for HPLC because many compounds exist in two mirror-image forms, or enantiomers, and separating and quantifying these enantiomers accurately is crucial for many industries such as pharmaceuticals and agrochemicals.

2. How is chiral purity determined using HPLC?

Chiral purity is determined using HPLC by separating the enantiomers using a chiral stationary phase column, which has a specific molecular structure that interacts differently with each enantiomer. The separated enantiomers are then detected and quantified using a detector, such as a UV or mass spectrometer.

3. What factors can affect the chiral purity results in HPLC?

Several factors can affect chiral purity results in HPLC, including the type and quality of the chiral stationary phase column, the composition and flow rate of the mobile phase, and the detection method used. Additionally, sample preparation and handling, as well as the expertise of the analyst, can also impact the results.

4. What is the typical range of chiral purity values obtained in HPLC?

The typical range of chiral purity values obtained in HPLC is between 90-99%, with a higher percentage indicating a higher enantiomeric excess. However, it is essential to note that the acceptable range can vary depending on the specific application and industry.

5. How can one improve the chiral purity results obtained in HPLC?

To improve chiral purity results in HPLC, one can use a high-quality chiral stationary phase column, optimize the mobile phase composition and flow rate, and ensure proper sample preparation and handling. Additionally, using a more sensitive detection method and having a well-trained analyst can also lead to improved results.

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