In the first place, general explanation of quantitative ultramicroanalyses will be given, and the basic procedures such as blood collection, pipetting, etc, are discussed. Next the future problems will be stated and finally actual examples of the analyses which were made in this hospital are displayed. (1) Explanation of quantitative ultramicroanalyses.
1. In the field of clinical chemistry, quantitative ultramicroanalyses are defined as an assay method which is carried out by using samples of 0.1ml or less, and routine tests have been already practised on this scale, and its necessity has gradually come to be widely appreciated.
2. There are various methods for this analyses, and the most ideal one is micromacro method which is done by using sample of ultramicro quantity and reagent of regular quantity.
3. In order to obtain a result of high precision, it is desirable that deproteiniztion should be avoided and the whole process must be completed in one test tube.
(2) From blood collection to separation of serum.
1. When the blood is collected from the sole of a foot by a capillary tube, the value of the chemical composition of the serum is almost the same as that of blood collected from cubital vein by an ordinary syringe.
2. As to potassium and lactate dehydrogenase, in most cases, the value is rather high in the blood collected by a capillary tube, and it is assumed that hemolysis and also mixing in of tissue fluid cause this.
3. Even in the case of blood collected by a capillary tube, the value of blood sugar became gradually low by the time of separation of serum unless the capillary tube had been coated with sodium fluoride. (3) Pipetting
1. It is desirable to collect sample and standard solution with the same pipet, and the viscosity and temperature should also be the same.
2. The contents of the pipet should be so completely blown out that no droplet is left on the inner surface of the pipet and one must be careful about the speed of blowing and using a clean pipet.
(4) In case of colorimetry.
When the amount of sample used for colorimetry is small and still a measurement of high sensitivity must be obtained:
1. Use microcuvette devised not to shorten the light path.
2. It is necessary to measure with light of narrow band width. (5) Future problems in quantitative ultramicroanalyses.
Generally speaking, as ultramicroanalyses progress, the precision of the test tends to drop. But this problem can be overcome by improving the method and mechanizing the tools and processes. The ideal aim is in the complete automation of ultramicroanalyses, and for the time being, our hospital introduced semiautomatic quantitative ultramicroanalyses of urea nitrogen and alkaline phosphatase using EEL Auto Chemist.
(6) Examples of quantitative ultramicroanalyses. Also the outline of examples of quantitative ultramicroanalyses which were discussed in our hospital were given: they were: Lactic acid (enzymatic colorimetry) Pyruvic acid (enzymatic method) Total bilirubin (fluorometry) Galactose (indirect and direct enzymatic method) Indocyanine green (ultramicro-colorimetry) Fibrinogen (nephelometry)
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