Chap H J, Moatti J P, Mieusset R, Nieto M, Laneelle G, Bennet P J, Mansat A, Pontonnier F, Douste-Blazy L
Laboratoire de Biochimie III, Hôpital La Grave, Toulouse, France.
Clin Chem. 1988 Jan;34(1):106-9.
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 89% 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) mumol 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 specificity, reliability, and convenience, allowing its use in routine evaluation of male fertility.
我们设计了一种新的酶法来测定人类精浆中的sn-甘油-3-磷酸胆碱(GPC)。该方法基于磷酸二酯酶(PDE)对GPC的水解作用,然后通过胆碱氧化酶法测定游离胆碱。整个过程包括在胆碱氧化酶和过氧化氢酶存在的情况下进行第一次孵育,以消除精浆中存在的过量胆碱(与GPC相比多10倍)。每次测定中,吸光度和浓度在至少100 nmol范围内呈线性关系,分析回收率在89%至105%之间,在最高底物浓度下,批内和批间变异系数分别为3.2%和5.6%。使用该方法,我们发现21名生育男性的精浆中每份射精量含有5.22(标准差3.33)μmol,与之前通过色谱法获得的报告值在同一范围内。在输精管结扎术后,对10名志愿者进行测定,发现精浆中的GPC降至其原始值的28%。因此,这种新方法具有特异性、可靠性和便利性等合适的特性,可用于男性生育能力的常规评估。