What HPLC method are you using (chromatographic conditions)? Polysorbate 80 is a polymer.

Column – waters symmetry C18 (150x4.6)mm, 5μ

 Flow rate – 1ml/min

 Detection – By UV at 220nm

 Column oven Temperature – 25°C

 Sample cooler Temperature – 25°C

 Gradient programme

We followed a literature method

What is the method (reference)?  What is the sample matrix?

sample matrix is oral solid tablet. Method mentioned earlier. What the details you want?

Reference paper.  Almost everything absorbs at 220 nm so an attached chromatogram would be nice too.  What is the expected concentration of polysorbate 80?

dear Sarkar,

1. Please check sample preservation if your sample is in liquid form then solvent may be evaporate 

2.. Calibrate the Instrument

3. Check Conditioning properly

Good Luck

Dear Anindita.

Kindly share literature method as i am also struggling for Estimation in Cream