Analyhca ChrmtcaActa, 213 (1993) 443441 Elsevler Science Pubhshers B V , Amsterdam 443 Determination of inert sugars in urine by liquid chromatography with pulsed amperometric detection S K Sanghl ’ and W Th Kok Laboratory for Analytxal Chemrstry, Unrversrty of Amsterdam, Nteuwe Achtergracht 166, 1018 WVAmsterdam (Netherlands) G C M Koomen and F J Hoek Department of Cluucal Chemutry, Academu: MedEal Centre, Metirgdreef 9,lIM AZ Amsterdam (Netherlands) (Received 29th June 1992) Abstract bqmd chromatography and pulsed amperometnc detection with a gold workmg electrode have been used for the determmatlon of celloblose and rhamnose m urme samples A conventtonal sugar-separation column was used mth an acldlc mobile phase, and a potassmm hydronde solution was added post-column Under optimal conditions detection hmits of 10 ng were obtained The method could be used for the assessment of the human intestinal permeablhty by the determmatlon of the urinary excretion of mert sugars Lmuts of determmatton m unne were 5 mg 1-l for celloblose and 15 mg 1-l for rhamnose Good agreement was found with the results of alternative methods Keywords Amperometry, Llqmd chromatography, Intestinal permeablhty, Sugars, Urme Alterations of the permeability of the mtestme for inert compounds are an mdlcatlon of the mucosal abnormahtles observed m various dlsor- ders The determmatron of the urinary excretion rate of exogenous compounds after oral admmls- tratlon can be used m the diagnosis and follow-up of, for Instance, coehac disease and Crohn’s dls- ease [l] In particular, the determmatlon of the ratio of the excretion rate of mert monosaccha- ndes and disaccharides finds mcreasmg apphca- tlon m research and clmlcal practice [2-61 Although enzymatic methods, quantitative pa- per chromatography and gas chromatography Correspondence to W Th Kok, Laboratory for Analytical Chemistry, Umverslty of Amsterdam, Nleuwe Achtergracht 166,1018 WV Amsterdam (Netherlands) ’ On leave from the Department of Chermstry, Bhdal Instl- tute of Technology (Seth Balknshna Memonal), Bhlhu House, Durg, MP (India) (GC) are also used for the determmatlon of these sugars m urine samples, liquid chromatography (LC) 1s now the pnmary approach for this analy- SIS The separation can be performed on catlon- exchange columns especially designed for this purpose, or, alternatively, on anion-exchange columns operated with a high-pH mobile phase A third approach 1s to use a normal-phase [7,8] or reversed-phase [9,101 column for the separation after denvatlzatlon of the sugars When the often troublesome post- or precol- umn denvatlzatlon techniques are to be avoided, until recently a refractive index (RI) detector had to be used for the detection of (underrvatlzed) sugars m LC However, for complex samples such as urine, the sensltmty and selectivity of this detector 1s m many cases msufflclent A break- through m this field was achieved with the devel- opment of the pulsed amperometrlc detection (PAD) technique, usmg platmum [11,12] or gold 0003-2670/93/$06 00 0 1993 - Elsemer Science Publishers B V All nghts reserved