Zelikman I, Hjertén S
Institute of Biochemistry, Uppsala University, Sweden.
Biomed Chromatogr. 1989 Jul;3(4):161-5. doi: 10.1002/bmc.1130030405.
A new method based on displacement electrophoresis has been developed for the determination of the total and free concentration of propranolol, a beta-adrenergic blocker drug, in plasma. To determine the total concentration the drug is extracted from human plasma into a chloroform + heptane mixture in the presence of ammonia. After evaporation of the solvent mixture the residue is submitted to displacement electrophoresis in a glass tube containing a two-layer polyacrylamide gel. When the propranolol electrophoretically leaves the gel column it is transferred by a buffer flow to the cuvette of a fluorimeter for continuous detection and quantification. The concentration of the free non-protein bound drug can be determined by the same displacement electrophoresis technique following extraction of the plasma sample into the above organic solvent mixture in the absence of ammonia. Alternatively the extraction procedure can be exchanged for dialysis for 1 h. To decrease the risk that large size material might comigrate with propranolol two layers of polyacrylamide are used, one having small pores to retard such material. In addition, we use fluorimetric detection which means that many possible contaminants are not recorded and therefore do not affect a quantitative determination of propranolol.
已开发出一种基于置换电泳的新方法，用于测定血浆中β-肾上腺素能阻滞剂药物普萘洛尔的总浓度和游离浓度。为了测定总浓度，在氨存在的情况下，将药物从人血浆中萃取到氯仿+庚烷混合物中。溶剂混合物蒸发后，将残留物在装有两层聚丙烯酰胺凝胶的玻璃管中进行置换电泳。当普萘洛尔通过电泳离开凝胶柱时，它会被缓冲液流转移到荧光计的比色皿中进行连续检测和定量。游离非蛋白结合药物的浓度可以通过相同的置换电泳技术来测定，即将血浆样品在无氨的情况下萃取到上述有机溶剂混合物中。或者，萃取程序可以换成透析1小时。为了降低大尺寸物质可能与普萘洛尔一起迁移的风险，使用了两层聚丙烯酰胺，其中一层具有小孔以阻滞此类物质。此外，我们使用荧光检测，这意味着许多可能的污染物不会被记录，因此不会影响普萘洛尔的定量测定。
