Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 409-418 |
| Number of pages | 10 |
| Journal | Free Radical Biology and Medicine |
| Volume | 34 |
| Issue number | 4 |
| DOIs | |
| State | Published - Feb 15 2003 |
| Externally published | Yes |
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Keywords
- 2,3-dinor-8-iso-prostaglandin F
- 8-Iso-prostaglandin F
- Free radicals
- Human urine
- LC-MS/MS
- Oxidative stress
- Quantification
ASJC Scopus subject areas
- Biochemistry
- Physiology (medical)
Cite this
Quantification of 8-iso-prostaglandin-F2α and 2,3-dinor-8-iso-prostaglandin-F2α in human urine using liquid chromatography-tandem mass spectrometry. / Liang, Yuanling; Wei, Ping; Duke, Russell W.; Reaven, Peter D.; Harman, S. Mitchell; Cutler, Richard G.; Heward, Christopher B.
In: Free Radical Biology and Medicine, Vol. 34, No. 4, 15.02.2003, p. 409-418.Research output: Contribution to journal › Article
}
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.
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
UR - http://www.scopus.com/inward/record.url?scp=0037441349&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037441349&partnerID=8YFLogxK
U2 - 10.1016/S0891-5849(02)01018-3
DO - 10.1016/S0891-5849(02)01018-3
M3 - Article
C2 - 12566066
AN - SCOPUS:0037441349
VL - 34
SP - 409
EP - 418
JO - Free Radical Biology and Medicine
JF - Free Radical Biology and Medicine
SN - 0891-5849
IS - 4
ER -