Simple, rapid enzymatic determination of glycerophosphocholine in human seminal plasma

Abstract

We have devised a new enzymatic determination of sn-glycero-3-phosphocholine (GPC) in human seminal plasma. This is based on GPC hydrolysis by a phosphodiesterase (PDE), free choline being then determined by the choline oxidase method. The whole procedure involves a first incubation in the presence of choline oxidase and catalase, to eliminate the excess of choline present in seminal plasma (10-fold, compared with GPC). Absorbance and concentration are linearly related up to at least 100 nmol per assay, analytical recovery ranges between 98% and 105%, and intra- and interassay CVs are 3.2% and 5.6%, respectively, at the highest substrate concentration. Using this procedure, we found seminal plasma from 21 fertile men to contain 5.22 (SD 3.33) μmol per ejaculate-within the same range as previously reported values obtained chromatographically. After vasectomy, GPC in seminal plasma decreased to 28% of its original value, as determined in 10 volunteers. Thus this new method displays appropriate characteristics of specifity, reliability, and convenience, allowing its use in routine evaluation of male fertility.