thiobarbituric-acid and Infertility--Male

A simple and reliable high performance liquid chromatographic (HPLC) method has been developed and validated for the analysis of malondialdehyde (MDA) in human seminal plasma. After human seminal plasma was hydrolyzed, MDA, one of the hydrolysis products, reacted with thiobarbituric acid (TBA) to form MDA (TBA)2, a red-colored adduct with a maximum absorbance at 532 nm. HPLC separation of the adduct in human seminal plasma was performed on a Lichrospher C18 column. A mobile phase composed of 0.025 mol/L KH2PO4 (pH 6.2)-methanol in 58:42 (v/v) was found to be the most suitable ratio for this separation at a flow rate of 1.0 mL/min and enabled the baseline separation of the adduct with isocratic elution. Under the chromatographic conditions described, the MDA-TBA adduct had a retention time of approximately 4 min, and good separation and detectability of MDA in human seminal plasma samples were obtained. The method proved to be linear in the range of MDA from 0.10 micromol/L to 2.50 micromol/L. The relative standard deviations of MDA analysis within- and between-assay were 3.1% (n = 7) and 3.8% (n = 5), respectively. The average recoveries were 90.0% -98.8% for the human seminal plasma samples. The method has been successfully applied to the study of male infertility induced by overproduction of lipid peroxidation in male reproductive system. Exception of obstructive azoospermic group, MDA concentrations of seminal plasma in control group made very significant difference from those in other infertile groups (P < 0.01).

Topics: Adult; Chromatography, High Pressure Liquid; Humans; Infertility, Male; Lipid Peroxidation; Male; Malondialdehyde; Semen; Thiobarbiturates

This study was designed to investigate the relationship between the effects of testicular reactive oxygen species (ROS) levels and testicular histology on infertile patients with the aid of xanthine oxidase system and testicular tissue malondialdehyde levels. Forty patients with idiopathic infertility constituted our study group. Bilateral testicular biopsies were performed and spermatogenesis was assessed histopathologically. Patients were divided into 4 groups according to spermatogenic pattern (normal spermatogenesis; hypospermatogenesis; maturation arrest; Sertoli cell only syndrome). Testicular tissue xanthine oxidase and malondialdehyde (MDA) concentrations were analyzed in each sample by spectrophotometric assay and thiobarbituric acid reaction assay, respectively. Testicular tissue MDA and xanthine oxidase concentrations were not statistically different in patients having normal spermatogenesis, with respect to Sertoli cell only syndrome, maturation arrest and hypospermatogenesis, respectively. As a result of our study we think that there are still some factors other than ROS which may be important contributors to spermatogenetic injury that need to be examined.

Topics: Adult; Biopsy; Humans; Infertility, Male; Male; Malondialdehyde; Reactive Oxygen Species; Spectrophotometry; Spermatogenesis; Testis; Thiobarbiturates; Xanthine Oxidase