Ascorbic Acid in human seminal plasma: determination and its relationship to sperm quality

Abstract

The objective of the present study was to assess the ascorbic acid (AA) levels in seminal plasma of the fertile and infertile men and to investigate its relationship with sperm count, motility and normal morphology. Semen samples were provided by fertile [smoker (n = 25), nonsmoker (n = 21)] and infertile men [smoker (n = 23), nonsmoker (n = 32)]. A simplified method of reverse phase high performance liquid chromatography (RP-HPLC) procedure using UV detection was applied for the determination of seminal AA. Fertile subjects, smoker or not, demonstrated significantly higher seminal AA levels than any infertile group (p<0.01). Nonsmokers had high, but no significant, mean AA levels in their seminal plasma compared with smokers. Seminal AA in fertile and infertile (smokers or nonsmokers) males correlated significantly with the percentage of spermatozoa with normal morphology (p<0.01). Seminal AA decreased significantly in infertile men. Decrease of seminal plasma AA is a risk factor for low normal morphology of spermatozoa and idiopathic male infertility. Measurement of seminal AA in the seminal plasma of males with a history of subfertility or idiopathic infertility is necessary and can be helpful in fertility assessment.

Keywords: RP-HPLC; ascorbic acid; male infertility; seminal plasma; sperm quality.

Fig. 1

Representative chromatograms from ascorbic acid (AA) in seminal plasma. Samples were prepared as described in materials and methods. The mobile phase consisted of 2.5% acetic acid glacial and 80% methanol in ddH₂O; flow rate: 1 ml/min and spectrophotometric detection at 254 nm; column: a Spherisorb C18 column (150 × 4.6 mm, 5 µm particle size).

Fig. 2

Correlation between the seminal plasma AA content with sperm with normal morphology. Seminal plasma AA was positively correlated with sperm with normal morphology (r² = 0.0796, p<0.01). The Pearson correlation test was used to analyze and examine the relationship between AA with sperm parameters.