HCID 25444, compound → Cholesterol

HRID 1292723Notes

Reaction equation

HRID 1292723 reaction equation
[CH3:1][C@H:2]1[CH2:85]O[C@@:5]2(O[C@H:8]3[C@@H:10](O)[C@H:11]4[C@@H:16]5[CH2:17][CH2:18][C@H:19]6[CH2:24][C@@H:23]([O:25][C@@H]7O[C@H](CO)[C@H](O[C@@H]8O[C@H](CO)C(O)[C@H](O[C@@H]9OC[C@@H](O)[C@H](O)[C@H]9O)[C@H]8O[C@@H]8O[C@H](CO)[C@H](O)[C@H](O[C@@H]9O[C@H](CO)[C@@H](O)[C@H](O)[C@H]9O)[C@H]8O)[C@H](O)[C@H]7O)[C@H:22](O)[CH2:21][C@:20]6([CH3:80])[C@H:15]5[CH2:14][CH2:13][C@:12]4([CH3:81])[C@H:7]3[C@@H:6]2[CH3:83])[CH2:4][CH2:3]1.C1(=O)OC(=O)C=C1.C(N1CCCC1=O)=C>>[CH3:85][CH:2]([CH2:3][CH2:4][CH2:5][C@H:6]([C@@H:7]1[C@:12]2([CH3:81])[C@H:11]([C@H:16]3[C@H:15]([CH2:14][CH2:13]2)[C@:20]2([CH3:80])[C:19]([CH2:24][C@H:23]([CH2:22][CH2:21]2)[OH:25])=[CH:18][CH2:17]3)[CH2:10][CH2:8]1)[CH3:83])[CH3:1]

Participants

3

Reactant 2

HCID 25444HCID 25444, compoundHCID 22904624HCID 22904624, compound

Product 1

HCID 5997Cholesterol

Procedure

The sticks were each placed into 100 uL solution of an A-C-B reagent in wells of a 96 well microwell plate. The reagent was a conjugate of digitonin (A) linked to horseradish peroxidase (B) through a maleic anhydride-N-vinylpyrrolidone copolymer (C) and was used at a concentration of approximately 1 μg/mL. The sticks were left in the solution for 15 minutes at room temperature, after which they were removed and placed into new wells of a microwell plate containing 200 μL of wash solution. The microwell plate was agitated to effect washing and after 1 min the sticks were removed to new wells containing 200 μL of fresh wash solution and again agitated for 1 min. Washing with agitation was done a third time, after which the sticks were removed and placed in 100 uL of a substrate solution (Enhanced K-Blue reagent). The sticks were incubated with the substrate solution, in the dark, for 15 minutes at room temperature, and then removed. One hundred (100) μL of 1 N sulfuric acid were added to the wells with the substrate solution to stop further reaction and the optical density of the resulting solution was read at 450 nm on a plate reading spectrophotometer, to provide a measure of the amount of cholesterol in the skin sample.

Workup

  1. customafter which they were removed
  2. additioncontaining 200 μL of wash solution
  3. washto effect washing and after 1 min the sticks
  4. customwere removed to new wells
  5. additioncontaining 200 μL of fresh wash solution
  6. stirringagain agitated for 1 min
  7. washWashing with agitation
  8. customafter which the sticks were removed
  9. waitin the dark, for 15 minutes at room temperature
  10. customremoved
  11. additionOne hundred (100) μL of 1 N sulfuric acid were added to the wells with the substrate solution
  12. customreaction

Sources & evidence

ORD record
ord-e0882a063f6b42ae96f54bd9e7dedda6
Source dataset
uspto-grants-2007_07
Patent number
US07238494B2

Reaction notes

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