111 DESIGN AND ANALYSIS OF BIOLOGICAL ASSAYS
The potency of several Pharmacopeial drugs must be determined by bioassay. A controlling factor in assay design and analysis is the variability of the biological test system, which may vary in its mean response from one laboratory to another, and from time to time in the same laboratory. To control this type of variation, the response to a Pharmacopeial drug is compared with a USP Reference Standard or other suitable standard. For convenience, each such preparation will be called the Standard and each preparation under assay, or Sample, the Unknown, and these will be designated respectively by the symbols S and U. (The Sample is sometimes referred to as the test preparation.)
After elimination of extraneous variables from the comparison of the Standard and the Unknown, an error variance is computed from the remaining variation, which, while uncontrolled, can nevertheless be measured. The error variance is required in calculating the confidence interval of the assayed potency. The confidence interval, known also as the fiducial interval, is so computed that its upper and lower limits are expected to enclose the true potency of the Unknown in 19 out of 20 assays. Many assay procedures fix the acceptable width of the confidence interval, and two or more independent assays may be needed to meet the specified limit. The confidence limits of the individual component assays usually overlap.
The aim of this chapter is to present a concise account of biometrical procedures for the USP bioassays. Its various sections are interrelated. Although the procedures are planned primarily for the assay of a single Unknown, equations for the joint assay of several Unknowns are given in context throughout the chapter and are summarized in the last section. Proof that an assayed potency meets its required confidence limits may be based also upon other recognized biometric methods that have a precision equivalent to that of the methods outlined herein.
A glossary of the terms used in the equations is provided at the end of this chapter.
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USP35NF30 Page 106Pharmacopeial Forum: Volume No. 36(4) Page 952