[Determination of lipid peroxidation in human seminal plasma by high performance liquid chromatography and its diagnostic value of male infertility].
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).
Li
Ke
K
The Center of Medical Laboratory Science, Nanjing General Hospital of Nanjing Command, PLA, Nanjing 210002, China.
Shang
Xuejun
X
Chen
Yonggang
Y
D000328:Adult; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007248:Infertility, Male / Q000378:metabolism*; D015227:Lipid Peroxidation; D008297:Male; D008315:Malondialdehyde / Q000032:analysis*; D012661:Semen / Q000737:chemistry*; D013858:Thiobarbiturates / Q000032:analysis
D013858:Thiobarbiturates; D008315:Malondialdehyde; C029684:thiobarbituric acid