Simultaneous determination of phenolphthalein and phenolphthalein glucuronide from dog serum, urine and bile by high-performance liquid chromatography.
A procedure is described to simultaneously quantitate phenolphthalein and its glucuronide metabolite from dog serum, urine and bile using high-performance liquid chromatography. The major advantages of this over pre-existing methods include direct analysis of the parent compound and glucuronide metabolite without enzymatic hydrolysis, increased sensitivity and the potential for automation of a large number of samples. Analytes were extracted from serum and urine using a combination of liquid- and solid-phase extraction methodology. Bile samples were analyzed directly after a twenty-fold dilution with mobile phase. The components plus internal standard were separated by reversed-phase high-performance liquid chromatography using step gradient elution and quantitated by the absorbance of ultraviolet light at 230 nm. Limits of detection from 1 ml of serum, 0.1 ml of urine and 0.05 ml of bile were 0.1, 0.5 and 10 microgram/ml for phenolphthalein and 0.1, 10 and 50 microgram/ml for phenolphthalein glucuronide, respectively.
Wilhelm
J A
JA
Rutgers University, College of Pharmacy, Piscataway, NJ 08855.
Bailey
L C
LC
Shepard
T A
TA
Venturella
V S
VS
D000818:Animals; D001646:Bile / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D020113:Phenolphthalein; D010635:Phenolphthaleins / Q000378:metabolism*; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D010635:Phenolphthaleins; C016651:phenolphthalein glucuronide; D020113:Phenolphthalein