Ultramicro Methods in Biochemistry. I. General Considerations. II. Procedures for the Determination of Serum Bilirubin.

摘要

A battery of biochemical tests to survey the function of body organs and systems of animals under a variety of experimental circumstances was selected. The choice of procedures was guided by an analytical concept, often employed clinically, to reveal early failure of the integrative mechanisms responsible for maintaining homeostasis. More specifically, the methods were chosen to assess general metabolism; to reveal general tissue destruction; and to provide selected data concerning adrenal, thyroid and liver function as well as the status of liver, kidney and bone. The need for serial sampling necessitated the development of an ultramicro system to allow numerous determinations on a small sample of blood. The Sanz ultra-micro system was evaluated and various aspects of the associated instrumentation are described and considered. Ultramicro procedures for determining serum bilirubin utilizing both the diazo reaction and spectrophotometric analysis were investigated. The preparation of a bilirubin standard in aqueous protein solution was developed and evaluated for stability and reproducibility. The relevant diazo coupling and spectrophotometric procedures are described and results comparing the two analytical principles are recorded. Various necessary precautions applicable to these methods are set forth and the advantages and disadvantages of each are detailed and discussed.