Assay of Urinary Free Fucose by Fluorescence Labeling and High-Performance Liquid Chromatography

1992 ◽  
Vol 38 (5) ◽  
pp. 752-755 ◽  
Author(s):  
J Suzuki ◽  
A Kondo ◽  
I Kato ◽  
S Hase ◽  
T Ikenaka
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Cancer Patients ◽  
Healthy Subjects ◽  
High Performance ◽  
Fluorescence Labeling ◽  
Anion Exchange Column ◽  
Highly Sensitive ◽  
Cleanup Procedure ◽  
The Difference

Abstract The concentrations of free fucose and other sugars in urine of cancer patients and healthy subjects were analyzed by high-performance liquid chromatography. After the urine samples were dried, we coupled the sugars in the residue with 2-aminopyridine to be detected as fluorescent derivatives. We then analyzed the pyridylamino derivatives of the sugars with an anion-exchange column and borate buffer. The difference between cancer patients and healthy subjects for the mean concentrations of fucose corrected for creatinine was significant (P less than 0.001). We checked the relationship between the concentrations of other sugars and the presence of cancer. This method is highly sensitive, and neither a cleanup procedure before labeling nor purification before injection into the column is needed. Not only fucose but also other sugars can be detected simultaneously, so this method should be useful for studying any changes in sugars in urine in various diseases.

scholarly journals Genotyping of the Lactase-Phlorizin Hydrolase C/T-13910 Polymorphism by Means of a New Rapid Denaturing High-Performance Liquid Chromatography-Based Assay in Healthy Subjects and Colorectal Cancer Patients

2007 ◽  
Vol 12 (5) ◽  
pp. 733-739 ◽  
Author(s):  
Ada Piepoli ◽  
Enrico Schirru ◽  
Angela Mastrorilli ◽  
Annamaria Gentile ◽  
Rosa Cotugno ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Colon Cancer ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Cancer Patients ◽  
Healthy Subjects ◽  
High Performance ◽  
Genetic Variant ◽  
Increased Risk ◽  
Colorectal Cancer Patients

Adult-type hypolactasia results from the progressive decline of lactase-phlorizin hydrolase activity in enterocytes after weaning. Lactase nonpersistence may determine a primary lactose intolerance with reduced diary product consumption, which is possibly related to an increased risk of colon cancer. Recently, a genetic variant C/T—13910 upstream of the lactase-phlorizin hydrolase ( LCT) gene has been strongly correlated with the lactase persistence/nonpersistence trait in both family and case-control studies. The authors validate a denaturing high-performance liquid chromatography (dHPLC)—based assay versus conventional genotype sequencing in detecting the C/T—13910 polymorphism of LCT and evaluate its prevalence in 2 different Italian geographical areas and in colorectal cancer patients. DNA samples of 157 healthy subjects and 124 colon cancer patients from Apulia and of 97 healthy subjects from Sardinia were evaluated for the C/T—13910 polymorphism by dHPLC, sequencing, and restriction fragment length polymorphism (RFLP). Under optimized conditions, dHPLC was as sensitive as DNA sequencing and detected a new genetic variant (T/C-13913) in 2 individuals that was not identified by RFLP assay. Frequency of lactase nonpersistence genotype (C/C—13910) was similar in healthy subjects from 2 different Italian geographical areas and not increased in patients with colorectal cancer. The results indicate that the dHPLC method may be used as a rapid, noninvasive, and laborsaving screening tool for genotyping C/T—13910 polymorphism, with high success, low cost, and reproducibility. ( Journal of Biomolecular Screening 2007:733-739)

Differential intestinal deconjugation of taurine and glycine bile acid N-acyl amidates in rats

1992 ◽  
Vol 262 (2) ◽  
pp. G351-G358
Author(s):  
R. Zhang ◽  
S. Barnes ◽  
R. B. Diasio
Keyword(s):  
High Performance Liquid Chromatography ◽  
Small Intestine ◽  
Liquid Chromatography ◽  
Bile Acid ◽  
Chenodeoxycholic Acid ◽  
High Performance ◽  
Biliary Fistula ◽  
The Difference ◽  
The Stability ◽  
Rat Bile

Mechanisms responsible for the difference in the relative amounts of taurine- and glycine-conjugated bile acid N-acyl amidates (Tau/Gly ratio) are not fully understood. In the present study, the stability of taurine- and glycine-conjugated bile acid N-acyl amidates during intestinal transit and absorption was examined to investigate the contribution of intestinal deconjugation to the Tau/Gly ratio in rat bile. Radiolabeled chenodeoxycholic acid (CDC) and its N-acyl amidates with glycine (CDC-Gly) or taurine (CDC-Tau) were introduced into the lumen of the upper small intestine in the biliary fistula rats, and radioactive metabolites in bile, blood, urine, and tissues were identified and quantitated by high-performance liquid chromatography. Results indicated that 1) extensive deconjugation of CDC-Gly occurs during intestinal absorption; 2) CDC-Tau is recovered in bile largely intact; and 3) newly synthesized CDC-Tau and CDC-Gly are formed in a ratio of less than 2:1 after administration of [14C]-CDC. In summary, the present study demonstrates that resistance of taurine-conjugated bile acid N-acyl amidates to hydrolysis in the intestine, rather than a difference in synthesis of taurine- and glycine-conjugated N-acyl amidates in liver, may account for the high Tau/Gly ratio in rat bile.

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