TY - JOUR
T1 - Quantification of 8-iso-prostaglandin-F2α and 2,3-dinor-8-iso-prostaglandin-F2α in human urine using liquid chromatography-tandem mass spectrometry
AU - Liang, Yuanling
AU - Wei, Ping
AU - Duke, Russell W.
AU - Reaven, Peter D.
AU - Harman, S. Mitchell
AU - Cutler, Richard G.
AU - Heward, Christopher B.
N1 - Funding Information:
We are deeply grateful to Dr. John Sperling and the Kronos Group for financial support. We are very grateful to Dr. L. Jackson Roberts and Dr. Jason Morrow, who provided invaluable help on this project; especially by performing GCMS tests on the urine samples used in the correlation studies. We also thank the staff of the Kronos Science Laboratory for technical support, and the Carl T. Hayden VAMC for their support of Dr. Reaven.
PY - 2003/2/15
Y1 - 2003/2/15
N2 - Quantification of 8-iso-prostaglandin F2α (8-iso-PGF2α) has been suggested to be a reliable indicator of lipid peroxidation that may be related to in vivo free radical generation, oxidative damage, and antioxidant deficiency. We have developed a LC-MS/MS method to quantify 8-iso- PGF2α and its dinor metabolite, 2,3-dinor-8-iso-prostaglandin F2α (2,3-dinor-8-iso-PGF2α), in human urine samples. After an initial purification step using an automated C18 solid phase extraction procedure, the urine sample was injected directly into a liquid chromatography (LC) system and detected with tandem mass spectrometry. The detection limit of the assay was 9 pg for 8-iso-PGF2α and 3 pg for 2,3-dinor-8-iso-PGF2α with both inter- and intraday variations of less than 12%. The inaccuracies were less than 3% for both analytes at three different levels. The urinary excretion rate of 2,3-dinor-8-iso-PGF2α was higher than that of 8-iso-PGF2α, and changed in proportion to the parent compound (R = 0.70, n = 60). Values obtained with this method showed good linear correlation to duplicate 8-iso-PGF2α measurements performed with GCMS (R = 0.97, n = 15). The mean excretion rates of 8-iso-PGF2α and 2,3-dinor-8-iso-PGF2α were significantly higher in smokers than in nonsmokers (0.53 ± 0.37 vs. 0.25 ± 0.15 μg/g creatinine, p = 0.002 for 8-iso-PGF2α and 8.9 ± 3.8 vs. 4.6 ± 2.6 μg/g creatinine, p = 0.003 for 2,3-dinor-8-iso-PGF2α, respectively). The excellent accuracy, reproducibility, and high throughput of this method should permit it to be used in large clinical studies and standard clinical laboratories.
AB - Quantification of 8-iso-prostaglandin F2α (8-iso-PGF2α) has been suggested to be a reliable indicator of lipid peroxidation that may be related to in vivo free radical generation, oxidative damage, and antioxidant deficiency. We have developed a LC-MS/MS method to quantify 8-iso- PGF2α and its dinor metabolite, 2,3-dinor-8-iso-prostaglandin F2α (2,3-dinor-8-iso-PGF2α), in human urine samples. After an initial purification step using an automated C18 solid phase extraction procedure, the urine sample was injected directly into a liquid chromatography (LC) system and detected with tandem mass spectrometry. The detection limit of the assay was 9 pg for 8-iso-PGF2α and 3 pg for 2,3-dinor-8-iso-PGF2α with both inter- and intraday variations of less than 12%. The inaccuracies were less than 3% for both analytes at three different levels. The urinary excretion rate of 2,3-dinor-8-iso-PGF2α was higher than that of 8-iso-PGF2α, and changed in proportion to the parent compound (R = 0.70, n = 60). Values obtained with this method showed good linear correlation to duplicate 8-iso-PGF2α measurements performed with GCMS (R = 0.97, n = 15). The mean excretion rates of 8-iso-PGF2α and 2,3-dinor-8-iso-PGF2α were significantly higher in smokers than in nonsmokers (0.53 ± 0.37 vs. 0.25 ± 0.15 μg/g creatinine, p = 0.002 for 8-iso-PGF2α and 8.9 ± 3.8 vs. 4.6 ± 2.6 μg/g creatinine, p = 0.003 for 2,3-dinor-8-iso-PGF2α, respectively). The excellent accuracy, reproducibility, and high throughput of this method should permit it to be used in large clinical studies and standard clinical laboratories.
KW - 2,3-dinor-8-iso-prostaglandin F
KW - 8-Iso-prostaglandin F
KW - Free radicals
KW - Human urine
KW - LC-MS/MS
KW - Oxidative stress
KW - Quantification
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U2 - 10.1016/S0891-5849(02)01018-3
DO - 10.1016/S0891-5849(02)01018-3
M3 - Article
C2 - 12566066
AN - SCOPUS:0037441349
SN - 0891-5849
VL - 34
SP - 409
EP - 418
JO - Free Radical Biology and Medicine
JF - Free Radical Biology and Medicine
IS - 4
ER -