Measurement of 20-Hydroxyeicosatetraenoic Acid in Human Urine by Gas Chromatography–Mass Spectrometry

Rivera, Jennifer; Ward, Natalie C.; Hodgson, Jonathan M.; Puddey, Ian B.; Falck, John R.; Croft, Kevin D.

doi:10.1373/clinchem.2003.025775

Cited by 46 publications

(28 citation statements)

References 11 publications

(13 reference statements)

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“…The assay was a modification of our previously described method [16] and extracted a smaller volume of urine (0.5 ml) and used 2 ng of d 6 -20-HETE (Cayman Chemical Co., Ann Arbor, Michigan, USA) as an internal standard. Plasma-free 20-HETE was measured in 125 ml of plasma.…”

Section: Methodsmentioning

confidence: 99%

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n-3 fatty acids reduce plasma 20-hydroxyeicosatetraenoic acid and blood pressure in patients with chronic kidney disease

Barden

Burke

Mas

et al. 2015

Journal of Hypertension

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Section: Methodsmentioning

confidence: 99%

“…The column was then washed with methanol/water (50/50) and dried under vacuum (6 mmHg for 2 min) before elution of 20-HETE with hexane/ethyl acetate/acetic acid (75/25/1). The sample was dried and derivatized as previously described for urine [16].…”

Section: Methodsmentioning

confidence: 99%

n-3 fatty acids reduce plasma 20-hydroxyeicosatetraenoic acid and blood pressure in patients with chronic kidney disease

Barden

Burke

Mas

et al. 2015

Journal of Hypertension

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“…17 Briefly, an internal standard, [2H 2 ]-20-HETE (2 ng) was added to all of the urine samples (2 mL) before incubation with glucuronidase to free all of the conjugated 20-HETE. After hydrolysis, samples were extracted using Bond Elut-Certify II cartridges before high-performance liquid chromatography purification, formation of the tBDMS-PFB derivative, and finally gas chromatography/ mass spectrometry analysis in select ion monitoring mode using negative chemical ionization.…”

Section: Analysis Of 20-hetementioning

confidence: 99%

A Single Nucleotide Polymorphism in the CYP4F2 but not CYP4A11 Gene Is Associated With Increased 20-HETE Excretion and Blood Pressure

et al. 2008

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Abstract-Arachidonic acid is a major fatty acid that can be metabolized by the cytochrome P450 enzyme to a number of bioactive eicosanoids. A major metabolite of this oxidation is 20-hydroxyeicosatetraenoic acid, which acts as a potent vasoconstrictor. However, in the kidney, its vasoconstrictor actions can be offset by its natriuretic properties. A guanine-to-adenine polymorphism in the CYP4F2 gene was associated with a reduction in 20-hydroxyeicosatetraenoic acid production in vitro. A thymidine-to-cytosine polymorphism in the CYP4A11 gene reduced catalytic activity by Ͼ50% in vitro and was associated with hypertension. The aim was to determine whether these 2 mutations are associated with urinary 20-hydroxyeicosatetraenoic acid excretion and blood pressure in humans. For the CYP4F2, 51% were homozygous for the G allele, 40% were carriers, and 9% were homozygous for the A allele. For CYP4A11, 72% were homozygous for the T allele, 25% were carriers, and 3% were homozygous for the C allele. The CYP4F2 GA/AA genotype was significantly associated with an increase in both 20-hydroxyeicosatetraenoic acid excretion and systolic blood pressure. The CYP4A11 CC/TC genotype was significantly associated with a reduction in 20-hydroxyeicosatetraenoic acid excretion but was not associated with blood pressure. We have demonstrated for the first time in humans that polymorphisms of the CYP4F2 and CYP4A11 genes have opposite effects on 20-hydroxyeicosatetraenoic acid excretion. The positive association between the CYP4F2 GA/AA genotype and both systolic blood pressure and 20-hydroxyeicosatetraenoic acid excretion strengthens a role for 20-hydroxyeicosatetraenoic acid in the modulation of blood pressure. A rachidonic acid is a major membrane fatty acid that can be metabolized by the cytochrome P450 (CYP450) enzymes to a range of bioactive compounds. These compounds are thought to play a central role in the regulation of blood pressure (BP), vascular tone, and renal function. 1,2 Within the vasculature, the CYP450 enzymes belonging to the 2-gene family (CYP 2B, 2C8, 2C9, 2C10, and 2J2) are responsible for the production of epoxides, whereas the -hydroxylases belonging to the CYP 4A and 4F families are involved in the production of hydroxyeicosatetraenoic acids (HETEs). 3,4 Animal studies have previously shown that disruption of the murine Cyp450 4a14 gene results in hypertension, possibly via increased expression of Cyp4a12. 5 20-HETE has been shown to play a role in vasoconstriction and renal salt handling in the spontaneously hypertensive rat. 2,6 -8 In humans, 20-HETE has been shown to play a role in regulation of natriuresis in salt-sensitive and salt-resistant hypertension, 9 and we have previously demonstrated a significant association between urinary 20-HETE excretion and both hypertension and endothelial dysfunction. 10 Paradoxically, within the kidney, 20-HETE can have either prohypertensive or antihypertensive actions, depending on its site of production. In the renal tubule, 20-HETE inhibits tubular sodium reabsorpti...

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“…24 The treated urine samples were applied to preconditioned Bond Elut-Certify II columns (Varian), washed with 2 mL of methanol/water (1:1), and then collected with 2 mL of hexane/ethyl acetate/acetic acid (75:25:1) for eicosanoid analysis.…”

Section: Measurements Of Eicosanoidsmentioning

confidence: 99%

Altered Release of Cytochrome P450 Metabolites of Arachidonic Acid in Renovascular Disease

et al. 2008

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Abstract-The aim of the present cross-sectional study was to investigate whether activation of the renin-angiotensin system in renovascular disease affects the cytochrome P450 /-1 hydroxylase (20-hydroxyeicosatetraenoic acid ) and epoxygenase (epoxyeicosatrienoic acids [EETs]) pathways of arachidonic acid metabolism in vivo, each of which interacts with angiotensin II. Plasma concentration and urinary excretion of 20-HETE and EETs and their metabolites, dihydroxyeicosatrienoic acids, were measured in urine and plasma by mass spectrometry in 10 subjects with renovascular disease, 10 with essential hypertension, and 10 healthy normotensive subjects (control subjects), pair-matched for gender and age. Vascular and renal function were evaluated in all of the subjects. Plasma 20-HETE was highest in subjects with renovascular disease (median: 1.20 ng/mL; range: 0.42 to 1.92 ng/mL) compared with subjects with essential hypertension (median: 0.90 ng/mL; range: 0.40 to 2.17 ng/mL) and control subjects (median: 0.45 ng/mL; range: 0.14 to 1.70 ng/mL; PϽ0.05). Plasma 20-HETE significantly correlated with plasma renin activity in renovascular disease (r s ϭ0.67; nϭ10; PϽ0.05). The urinary excretion of 20-HETE was significantly lower in subjects with renovascular disease (median: 12.9 g/g of creatinine; range: 4.4 to 24.9 g/g of creatinine) than in control subjects (median: 31.0 g/g of creatinine; range: 11.9 to 102.8 g/g of creatinine; PϽ0.01) and essential hypertensive subjects (median: 35.9 g/g of creatinine; range: 14.0 to 72.5 g/g of creatinine; PϽ0.05). Total plasma EETs were lowest, as was the ratio of plasma EETs to plasma dihydroxyeicosatrienoic acids, an index of epoxide hydrolase activity, in renovascular disease (ratio: 2.4; range: 1.2 to 6.1) compared with essential hypertension (ratio: 3.4; range: 1.5 to 5.6) and control subjects (ratio: 6.8; range: 1.4 to 18.8; PϽ0.01). In conclusion, circulating levels of 20-HETE are increased and those of EETs are decreased in renovascular disease, whereas the urinary excretion of 20-HETE is reduced. Key Words: eicosanoids Ⅲ 20-HETE Ⅲ EETs Ⅲ DHETs Ⅲ renal artery stenosis Ⅲ hypertension Ⅲ angiotensin II I n the past 2 decades, the view that the cytochrome P450 (CYP) monooxygenase pathway of arachidonic acid (AA) metabolism affects blood pressure and contributes to the development of hypertension is supported by studies in several experimental models. 1,2 However, little information is available on possible contributions of CYP products to essential hypertension (EH) and human renal vascular hypertension. 3 The role of angiotensin II (Ang II) in the development of hypertension and ischemic nephropathy in the experimental model of the 2-kidney 1-clip hypertension is evident, because a fall in renal perfusion pressure triggers renin release. Vasoconstriction, vascular remodeling, glomerulosclerosis, and interstitial matrix deposition implicate an array of mediators that operate downstream from Ang II. 4 CYP monooxygenases are expressed in the renal vasculature and nephron, 5 gen...

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Measurement of 20-Hydroxyeicosatetraenoic Acid in Human Urine by Gas Chromatography–Mass Spectrometry

Abstract: Presence of fetal DNA in maternal plasma and serum. Lancet 1997;350: 485-7

Cited by 46 publications

References 11 publications

n-3 fatty acids reduce plasma 20-hydroxyeicosatetraenoic acid and blood pressure in patients with chronic kidney disease

n-3 fatty acids reduce plasma 20-hydroxyeicosatetraenoic acid and blood pressure in patients with chronic kidney disease

A Single Nucleotide Polymorphism in the CYP4F2 but not CYP4A11 Gene Is Associated With Increased 20-HETE Excretion and Blood Pressure

Altered Release of Cytochrome P450 Metabolites of Arachidonic Acid in Renovascular Disease

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