Flash Purification of Carbohydrates with puriFlash® 5.030 and Integrated ELSD

Instrumentation
Flash chromatography system: puriFlash^® 5.030 with pack iELSD
Column: puriFlash^® 50µm NH2 F0025

Author
Applications Team
Advion Interchim Scientific

Introduction
Carbohydrates are non-chromophoric compounds that often lead to flat signals when using UV detection only. In this application note, we show carbohydrate purification with ELSD and how such detection can help in purification.

Why use ELSD from Advion Interchim Scientific?

• • Detect chromophoric and non chromophoric compounds as carbohydrates
  • Low maintenance with Isopropanol make-up solvent for ELSD
  • IPA push the sample to the ELSD
  • System cleaning meanwhile processing to purification => almost maintenance freeAutomatic refill
  • Eliminates risk of signal saturation or non-detection
  • Allow to detect and collect low concentration compounds
  • Easy set up (no parameters to predict, easy to use)
  • Automatic gain to clearly see all compounds at the same time (Figure 1)
  • Low sample consumption 40µL/min
  • ELSD is a destructive detection mode, thank to automated spliter quantity send to detector is control
  • Low gas consumption (1 – 1.5L/min at 1-1.5bar)
  • Low temperature technology

  
  Figure 1: Automatic gain allows users to clearly see all compounds at the same time

  Method
  Experimental Set Up
  Flash chromatography system: puriFlash^® 5.030 with pack iELSD
  Column puriFlash^®: 50µm NH2 F0025
  Sample: D(-)Fructose 100mg
  Alpha (D)-Lactose 100mg
  Detector: ELSD 35°C
  UV: 254nm

  

  Results & Discussion
  ELSD Signal
  
  Figure 2: ELSD showing a good signal intensity and smoothing.

  UV Signal 254nm
  

  UV Signal SCAN 200-800nm
  

  Figure 3 & 4: Compounds are not visible with UV. Scan start to show low intensity for the first compounds, but not enough to get good collection.

  Conclusion
  Carbohydrates are non-chromophore compounds and can’t be detected by UV, it is necessary to use ELSD detector that provide good detection.

  The ELSD detector that AIS propose is capable of provide several benefits for user:

• no need to define gain depending on the loading
• Thanks to low temperature technology, it is possible to look more compounds (especially temperature sensitive or volatile)
• In this application we used water and 35°C was enough to remove water response and get flat signal.