Hydrxoxyproline interference
Posted: Fri Dec 01, 2017 12:35 pm
We have a contamination or interference for the hydrolysate sample of hydroxyproline analysis. We think it is the product that we use that is the problem, because we can see a peak in the retention time of the hydroxyproline in a method blank. We have an 30-40 fold overestimation of our hydrolysate sample. We can see a shoulder peak or a split peak in those samples. Beside a contamination or interference problems coming from my product, do you have any other suggestion why I have a peak of hydroxyproline in a method blank?
My method that I use is :
1. The acid hydrolysis with 6 N HCl + 1% phenol for 24 hours at 110°C.
2. SPE with the Dowex resin.
3. Analyze with the 1290 infinity HPLC-DAD with an OPA and FMOC derivatization.
Column : InfinityLab Poroshell HPH-C18 4.6 * 150 mm 2.7-micron
Mobile phase A : 10 mM Na2HPO4 7H20 and 10 mM Na2B4O7 10H20
Mobile phase B : ACN:MeOH:H20 (45:45:10).
My method that I use is :
1. The acid hydrolysis with 6 N HCl + 1% phenol for 24 hours at 110°C.
2. SPE with the Dowex resin.
3. Analyze with the 1290 infinity HPLC-DAD with an OPA and FMOC derivatization.
Column : InfinityLab Poroshell HPH-C18 4.6 * 150 mm 2.7-micron
Mobile phase A : 10 mM Na2HPO4 7H20 and 10 mM Na2B4O7 10H20
Mobile phase B : ACN:MeOH:H20 (45:45:10).