IntroDuCtIon Artificial insemination is one of the methods widely used in the treatment of male infertility [1-4]. Many childless couples prefer homologous artificial insemination in which a portion of split ejaculate containing rich and active spermatozoa was shown to give good results in such cases [5-8]. Furthermore, in cases of oli-gospermia it was the preferred choice. These outcomes prompted spilt ejaculation studies in order to understand semen details and the origin of each component in semen [9-12] The different features of two elements, calcium and magnesium , in the human reproductive system and in semen, indicate their prominent role in fertilization. The aim of the current study is to investigate the quality of semen in each split ejaculate in order to recommend a suitable split for artificial insemination as well as to identify the origin of calcium and magnesium in semen. MaterIalS anD MetHoDS All glassware and plasticware were cleaned thoroughly [13, 14] (Valsa et al. 2012a; 2012b). Thirty-one healthy adults belonging to the Province of Gujarat, aged of 20 to 35 years, volunteered for the study. Prior to sample collection, their consent was obtained and they were instructed to maintain abstinence for three to five days prior to semen sample collection[15]. They were further instructed to collect complete semen samples following masturbation in three splits. Each split was to be collected in a clean, numbered wide mouthed sterile bottle [16]. Samples were evaluated as per WHO [8] criteria. Seminal plasma was separated by centrifugation (10x2000 rpm) [17]. The pellet of spermatozoa and seminal plasma were kept under-20 o C until the estimation of calcium and magnesium. Calcium and magnesium was estimated by a colorimetric method [18]. A comparative study of colorimetry and atomic absorption spectrophotometry produced similar results in case of calcium and magnesium [14].
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