Citric Acid → Thromboxane

HRID 615598

Reaction equation

HRID 615598 reaction equation
[CH3:1][CH2:2][CH2:3][CH2:4][CH2:5][C@H:6](O)/[CH:7]=[CH:8]/[C@@H:9]1[C@@H:13]([CH2:14]/[CH:15]=[CH:16]\[CH2:17][CH2:18][CH2:19][C:20](O)=O)[C@H:12]2O[O:24][C@@H:10]1[CH2:11]2.C(O)(=O)CC(CC(O)=O)(C(O)=O)O>CCOCC>[CH3:20][CH2:19][CH2:18][CH2:17][CH2:16][CH2:15][CH2:14][C@@H:13]1[C@@H:9]([CH2:8][CH2:7][CH2:6][CH2:5][CH2:4][CH2:3][CH2:2][CH3:1])[O:24][CH2:10][CH2:11][CH2:12]1

Participants

4

Reactant 2

HCID 311Citric AcidHCID 445049Prostaglandin H2

Product 1

HCID 114873Thromboxane

Reagents, catalysts & solvents 1

HCID 3283Diethyl EtherSolvent

Procedure

To a 12×75 mm tube containing [1-14C] PGH2 (2 μg, 0.01 μCi or 0.05 μCi) and inhibitor (1 mM final assay concentration, in 10 μl of ethanol) in 225 μl of phosphate buffer (0.1M, pH 7.5), was added to an aliquot of guinea pig lung mocrosomes (25 μl, containing 400 μg protein) or an aliquot of human platelet microsomes (25 μl, containing ≅130 g protein). The solution was incubated in a 22° water bath for 2 min.; ether (1 ml) and citric acid (25 μMoles) were added, and the tube plunged into an acetone-dry ice bath. The ether extract was transferred to a 1.0 ml Reacti-vial, and the volume reduced under nitrogen to ≅50 μl.

Workup

  1. concentrationassay concentration
  2. additionin 10 μl of ethanol) in 225 μl of phosphate buffer (0.1M, pH 7.5), was added to an aliquot of guinea pig lung mocrosomes (25 μl, containing 400 μg protein)
  3. additionan aliquot of human platelet microsomes (25 μl, containing ≅130 g protein)
  4. customwas incubated in a 22°
  5. customfor 2 min.
  6. customthe tube plunged into an acetone-dry ice bath
  7. extractionThe ether extract

Sources & evidence

ORD record
ord-f987717a68094bb3ac24ef7c396c803e
Source dataset
uspto-grants-1985_02
Patent number
US04500540

Linked notes

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