Abstract: Theophylline can be determined with an analytical element and method which utilize the inhibition, by theophylline, of alkaline phosphatase activity on an appropriate substrate. The assay is carried out at a pH of 9 or less. The element comprises a porous spreading zone, a subbing zone and at least one other zone. The zones are in fluid contact with each other. The isoenzyme of alkaline phosphatase and a suitable substrate for the isoenzyme are located in different zones of the element. The subbing zone contains polyvinylpyrrolidone and a buffer for maintaining a pH of 9 or less.

Abstract: Theophylline can be determined with an analytical element and method which utilize the inhibition, by theophylline, of alkaline phosphatase activity on an appropriate substrate. The assay is carried out at a pH of 9 or less. The element comprises an absorbent carrier material, a suitable buffer and first and second zones in fluid contact. The isoenzyme of alkaline phosphatase and a suitable substrate for the isoenzyme are located in different zones of the element. The isoenzyme is present in one of the zones in an amount of at least about 100 I.U./m.sup.2. This level of isoenzyme reduces or eliminates bias caused by endogenous alkaline phosphatase and hemoglobin during an assay.

Abstract: Theophylline can be determined with an analytical element and method which utilize the inhibition, by theophylline, of alkaline phosphatase activity on an appropriate substrate. The assay is carried out at a pH of 9 or less. The element comprises a porous spreading zone and at least one other zone in fluid contact with the spreading zone. The isoenzyme of alkaline phosphatase and a suitable substrate for the isoenzyme are located in different zones of the element. The element also contains a buffer for maintaining the pH at 9 or less, substantially all of which is located in the spreading zone.

Abstract: A heterogeneous, competitive binding immunoassay is conducted with a dry analytical element. The immunoassay is useful for determining a ligand (e.g. a therapeutic drug) and includes contacting a finite area of the element spreading layer with a sample of a liquid in the presence of a labeled ligand analog and an immobilized receptor. An immobilized ligand-receptor complex is formed within the finite area. Simultaneously, the uncomplexed ligand migrates horizontally away from the immobilized complex which remains in the center of the finite area. At least five seconds after the completion of the contacting, the amount of immobilized complex is measured in the center of the finite area.