ObjectivesMale infertility is on the rise. Artificial insemination is an option in many cases like oligozoospermia or oligoasthenozoospermia. Homologous insemination is helpful for some couples in whom sperm count is low. This study was aimed to understand the most suitable portion of split ejaculate for insemination and also the level of calcium and magnesium in each split.Materials and methodsA total number of 31 normal and clinically healthy adults participated in this study. They were instructed to maintain abstinence for 2-5 days prior to sample collection in three splits. Each split was evaluated as if it were a whole sample, following WHO criteria. Seminal plasma was separated. Calcium and magnesium levels were measured in seminal plasma and spermatozoa of each split.ResultsThe split ejaculate study revealed that the first portion contained a higher number of spermatozoa with better motility than the 2nd and 3rd splits. Similarly, the level of calcium and magnesium in seminal plasma and spermatozoa was greater in the first split, followed by 2nd and 3rd splits.ConclusionThe presented split ejaculate study has shown the first ejaculate as superior in quality in terms of total sperm count and percentage of active motility. The first split of ejaculation is recommended for homologous artificial insemination in case of oligozoospermia and oligoasthenozoospermia. This study has also shown that a major portion of two important elements, calcium and magnesium, is contributed by the prostate gland.

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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].

A study was conducted to establish the level of calcium and magnesium in human semen, seminal plasma and spermatozoa. One hundred and sixteen healthy, fertile young men submitted sample for this study. Total sperm count and active motility showed a positive correlation. Both elements were found more outside spermatozoa. They maintained a positive correlation. Spermatozoa count did not show correlation with any of these two elements. There was a negative correlation in calcium inside and outside spermatozoa.

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