Chromatography Forum

Hello,

I am running a sugar analysis method by HPLC-RI using Waters High Performance Carbohydrate Column (silica bonded with trifunctional amino propylsilane).

My chromatographic conditions are: M.P= 80% ACN, column temp=30°C, flow rate = 1.0ml/min.

For sample prep, I extract 1 g of sample in 25%ACN by vortex mixing, heating at 80°C for 20 min then centrifuging and filtering.

I notice sometimes (not usually) the sucrose peak will split, but only in my sample injection. I suspect it has to do with the sample prep. Only sucrose splits while glucose, fructose, maltose and lactose peaks remain intact.

What causes sugar to split?

I was able to simply resolve this issue by changing the guard column

Indeed, an HPLC column that is fouled or with a void in it (e.g. your guard column) is a common reason for peak "splitting" as is sample overloading. Was it the first peak to elute? That would make a lot of sense.

For more information on this topic, please review this free article; "HPLC Peak Splitting. Common Reasons For It"; https://hplctips.blogspot.com/2016/09/h ... s-for.html