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1) In a Reverse Phase HPLC procedure, will the most polar components in the samp

ID: 633379 • Letter: 1

Question

1) In a Reverse Phase HPLC procedure, will the most polar components in the sample elute near the beginning or late part of the chromatogram? How does this compare to HILIC? 2a) Regarding HPLC, explain specifically why it is vital that the mobile phase (Carrier) is either held under a slight vacuum or is purged with helium gas before entering the HPLC column. 2b) An analysis of fatty acids is desired. These fatty acids have boiling points in excess of 275O C. The sample is extracted with methanol to obtain a solution that only contains the fatty acids. Which of the following Methods/Cases is not possible and why, & which is the best approach & why? Case I-The methanol extract is mixed with a compound that reacts with the fatty acids and adds a methyl group to the end of the fatty acid

Explanation / Answer

HILIC; a sub-technique within HPLC, this method separates analytes based on differences in their polarity. HILIC most often uses a bonded polar stationary phase and a mobile phase made primarily of acetonitrile with water as the strong component. helium gas is merged in the sample before entering the HPLC because of avoiding other gas presence, so that we will get the clear result. for the fatty acid study Case 2, should be used that is the use of HPLC, it will be better for this study than HILIC which is better for amino acids and protein study.

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