HPLC with ELSD

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Kingmaker
Kingmaker's picture
HPLC with ELSD

 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

Dr. Analytical
Dr. Analytical's picture
First, can you confirm that

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.

Kingmaker
Kingmaker's picture
 Hello Dr. Analytical

 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.

Kingmaker
Kingmaker's picture
 Dr. Analytical Please.....

 Dr. Analytical Please.....

dws123
dws123's picture
I have never worked on ELSD

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