Advancing monosaccharides as biomarkers: part I. Development of fluorophore-assisted carbohydrate-electrophoresis in Chironomus riparius.
Fluorophore-assisted carbohydrate-electrophoresis (FACE) was developed as a bioassay for environmental stressors in larval Chironomus riparius. This quantitative technique involved acid hydrolysis and 2-aminoacridone labeling of monosaccharides followed by carbohydrate gel electrophoresis. Methods for carbohydrate isolation from whole tissue homogenates as well as migration distances for 23 different monosaccharides and 4 disaccharides were established. Sensitivity of the technique (5 microg/ml saccharide) exceeded those of other detection methods. Results showed seven distinct bands in larvae. Four migrated distances similar to those of ribose, glucose, galactose, and fructose. Three proved to be alternative reaction products (ARP). Experiments determined that two of the ARPs were primarily from glucose and one was from glycogen. FACE allowed different saccharides from multiple larval samples to be analyzed in parallel. Effects of toxicants and diet on bioenergetics could be studied using this technique.
Bentivegna
Carolyn S
CS
Department of Biology, Seton Hall University, 400 South Orange Avenue, South Orange, NJ 07079, USA. bentivca@shu.edu
D000609:Aminoacridines; D000818:Animals; D001681:Biological Assay / Q000379:methods; D015415:Biomarkers / Q000032:analysis; D002241:Carbohydrates / Q000737:chemistry; D002683:Chironomidae / Q000502:physiology*; D004586:Electrophoresis / Q000379:methods*; D004785:Environmental Pollutants / Q000009:adverse effects*; D005456:Fluorescent Dyes; D006868:Hydrolysis; D007814:Larva; D012680:Sensitivity and Specificity
D000609:Aminoacridines; D015415:Biomarkers; D002241:Carbohydrates; D004785:Environmental Pollutants; D005456:Fluorescent Dyes; C072515:2-aminoacridone