I am working on phytochemistry and I am facing problem with development of method for Sarsasapogenin on HPLC. As we know Sarsasapogenin has very weak UV absorption so It cant be detected on UV detector.
So I am trying to detect It on ELSD ( Model: Varian 385 LC )but still i cant find any peak.
Can anybody help me please..??
Sarsasapogenin
First, can you confirm that your detector is working? Can you inject other molecules and get a peak?
For the Sarsasapogenin, please provide your separation conditions (column, mobile phase, flow, etc.). Do you know if it is actually eluting? This is a large, hydrophobic molecule and it will be highly retained on a C18 column.
Hello Dr. Analytical
My Detector is working properly because I detected other moleclules peak.
In case of Sarsasapogenin, I tried different seperation conditions.
Mobile phase: MeOH : H2O, CH3CN : H2O with different ratio
Column: Merck, C18 (250 x 4.6 mm)
Flow rate : 1.0 ml/min
Detector condition: Nebuliser: 25 --up to 55 ' C
Evoperation: 25--up to 85 'C
Nitrogen Gas: 1.0--up to 1.6 SLM
But still I am facing problem with peak detection.
Dr. Analytical Please.....
I have never worked on ELSD and hence have no idea of it. However, i would love to learn.
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used hplc