TY - JOUR
T1 - Spectrophotometric assay of phenolphthalein in biological fluids
AU - Morris, Sarah M.
AU - Powell, D. W.
N1 - Funding Information:
The authors wish to thank Nancy Love for preparation of manuscript and Dr. B. E. Statland for helpful discussions of the method. This investigation was supported by Research Grant AM 15350 from the National Institutes of Arthritis, Metabolism and Digestive Diseases and by a grant from Ex-Lax Pharmaceutical Company. Inc. Dr. Powell is the recipient of Research Career Development Award (AM70454) from the National Institutes of Arthritis, Metabolism and Digestive Diseases.
PY - 1979/6
Y1 - 1979/6
N2 - An assay for phenolphthalein in biological fluids has been developed utilizing methods previously applied to the assay of bromosulphalein and to the deconjugation of steroidal compounds in urine. Intestinal perfusate, serum, and urine samples containing phenolphthalein are deproteinized with acidified acetone, the samples dried, and the phenolphthalein redissolved in ethanol. Color is developed with 0.5 m glycine buffer, pH 12, and the samples read at 550 nm after blanking the spectrophotometer with one of the replicates to which acidic glycine buffer is added. To measure conjugated phenolphthalein in urine, the sample is incubated overnight with β-glucuronidase/arylsulphatase prior to phenolphthalein determination as noted above. This method gives an accurate assay of phenolphthalein to 10-5 m concentrations with coefficients of variation between 2 and 8% and with no resulting interference from hemoglobin or bilirubin.
AB - An assay for phenolphthalein in biological fluids has been developed utilizing methods previously applied to the assay of bromosulphalein and to the deconjugation of steroidal compounds in urine. Intestinal perfusate, serum, and urine samples containing phenolphthalein are deproteinized with acidified acetone, the samples dried, and the phenolphthalein redissolved in ethanol. Color is developed with 0.5 m glycine buffer, pH 12, and the samples read at 550 nm after blanking the spectrophotometer with one of the replicates to which acidic glycine buffer is added. To measure conjugated phenolphthalein in urine, the sample is incubated overnight with β-glucuronidase/arylsulphatase prior to phenolphthalein determination as noted above. This method gives an accurate assay of phenolphthalein to 10-5 m concentrations with coefficients of variation between 2 and 8% and with no resulting interference from hemoglobin or bilirubin.
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U2 - 10.1016/0003-2697(79)90757-7
DO - 10.1016/0003-2697(79)90757-7
M3 - Article
C2 - 453532
AN - SCOPUS:0018333847
SN - 0003-2697
VL - 95
SP - 465
EP - 471
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -