Retention time
Posted: Thu Aug 18, 2011 10:31 am
Hi everybody! I have the following problem: I am trying to identify Pentosidine in hydrolysed (6N HCl 24 h 110 °C) collagen samples using a Phenomenex Gemini C18 150x4.6 column. Mobile phase is 0.1% TFA and 0.1% in 100% CH3CN respectively. Gradient used ranges from 1 to 7% at 0.8ml/min in 30min. Collagen samples are dryed after acid hydrolysis and resuspended in 0.1% TFA. The Pentosidine standard is dissolved in 0.1% TFA. The retention time of the Pentosidine standard is 15min. Pentosidine in collagen samples comes at 17min. When I mix Pentosidine standard and collagen sample in a ratio of 1:1 the Pentosidine standard also elutes at 17min. Now my questions: Is it possible that remaining HCl has an influence on the retention time and if so what can I do about it? Many thanks for your help! Regards. Tillman