Determination of Amino Acids and Polyamines in Human Erythrocytes : Part 1. Fundamental Studies

1) A simple, rapid analytical method for amino acids and polyamines in erythrocytes within a blood volume of l ml with a quantitative limit of precision ranging from 1-10 p moles was established using high performance liquid chromatography (HPLC)

2) A capillary sampling system (C-S system) which consists of a propylene capillary tube for hematocrit measurement was adopted for treatment of the erythrocytes. After fillihg the tube with blood, it was treated with a variety of method.

3) The free amino acids in erythrocytes had such characteristics as (1) high levels of acidic amino acids, (2) low levels _of Val, Met, Ile, Phe, Trp and Arg and (3) branched chain amino acids (Val, Ile, Leu) had a tendency to readily pass through the erythrocyte membrane.

4) Washing of erythrocyte pellets with saline was not good for measurement of amino acids in erythrocytes, particularly the branched chain amino acids. The space rate of non-washed erythrocyte pellets was calculated as (free amino acid levels in blood pellets-space rate x free amino acid levels in plasma).

5) Polyamine level in erythrocytes changes in accordance with aging of erythrocytes, i.e. it begins decreasing from the time reticulocytes are released from the bone marrow and continues throughout the course of mature erythrocytes up to the time of destruction in the spleen.