Abstract
Duodenal fluid was collected using an Entero-testR string, which was swallowed at bedtime and removed the next morning. Fluid squeezed from the string was centrifuged 4 min and injected into a reverse-phase HPLC system equipped with a Guard-PAKR pre-column module and C-18 μBONDAPAKR column. Pigments were eluted with an acetonitrile gradient. Peaks were identified by: a) standards prepared from TLC of rat bile; b) in-vitro incubations of UDP-sugars (glucose, xylose, glucuronic acid) and bilirubin isomers (IIIα, IXα, XIIIα) with liver microsomes; and c) referral to Onishi's method (Biochem J 159:1899, 1980). Total bilirubin diglucuronide (DG), monoglucuronide (MG) and monoglucoside-monoglucuronide diester (DE, tentative assignment) account for more than 90% of the 436-nm peak areas. This has been defined as 100% in the following table; data = mean±SD.This method is safe, reproducible, requires no derivatization or extraction and is easily performed in the ambulatory setting. Bile pigment composition is similar in males and females and independent of estrus. Analysis of 4 male patients with Gilbert's syndrome (GS) demonstrates the increased excretion of MG seen in this condition.
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