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KMID : 1156220100360010044
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Journal of Environmental Health Sciences
2010 Volume.36 No. 1 p.44 ~ p.51
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Determination of 8-iso-PGF2α as Oxidative Stress Marker in Human Urine by High Performance Liquid Chromatography with Tandem Mass Spectrometry
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Kho Young-Lim
Lee Eun-Hee
Chae Hong-Jae
Choi Kyung-Ho
Pack Do-Myung
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Abstract
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This study aimed to develop analytical method for 8-isoprostanes as biomarkers for oxidative stress with LC/MS/MS technique and to apply the method for human urine samples. Analyzed compounds for urinary oxidative stress markers were 7 stereo-isomers of prostaglandins and the internal standard (iso-PGF2α-d₄) was used to adjust the recovery rate. The method for determining urinary iso-PGF2α consisted of solid phase extraction and LC/MS/MS detection. Separation of isomers of prostaglandins completed by porous graphitic carbon column and buffer solution. Detection limits for urinary markers of oxidative stress, iso-PGF2α with LC/MS/MS were 0.01 ng/㎖ by S/N ratio 3 and 0.028 ng/㎖ by calculated as to FDA method. The recovery (92.8~101.9%) and precision (8.8~20.7%) of analysis were feasible for detecting iso-PGF2α in real human urine samples. We detected 4 isomers of prostaglandins in human urine samples. Mean (standard deviation) of urinary iso-PGF2α concentration were 0.231 (0.117) ng/mg creatinine for smoking group and 0.154 (0.082) ng/㎎ creatinine for non-smoking group.
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KEYWORD
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oxidative stress, 8-iso-PGF2α, prostaglandin, LC/MS/MS
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