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- Posts: 17
- Joined: Mon Jun 27, 2022 1:20 pm
I'm analyzing samples to detect monosaccharides, organic acids and alcohols in HPLC. Unfortunately, the peaks for two primary products, ethanol (Eth) and butyric acid (HBu) tend to overlap while the retention time of other organic acids (e.g lactic acid) are bit too close to the RT of HBu and Eth for my comfort (1.5 min before HBu).
Can someone please tell me how I can separate the Eth and HBu peaks? Additionally, if there's something I can do to distance the lactic acid peak away from Eth/HBu that would be wonderful.
For reference, please find the specifications of the HPLC machine and the restrictions below:
1. HPLC: Shimadzu LC10A
2. Column: Rezex™ RHM-Monosaccharide H+ Ion Exclusion (300 x 7.8 mm)
3. Detector: RID (UV-Vis not possible)
4. Mobile Phase: Water (Reverse Osmosis)
5. Flow Rate: 0.5 ml/min [Isocratic Flow]
6. Column Temperature: 75 degree C (max. 85 C)
7. (Detector) Cell Temperature: 40 degree C
8. Injection Volume: 20 uL (Direct Injection)
9. (Detector) Response: 1.5 seconds
Sample Preparation
- 400uL of 0.02N H2SO4
- 300uL of sample (passed through 0.22um filter)
- 100uL of ISTD for organic acids (crotonic acid)
- 100uL of ISTD for alcohols (TBD)