Hydrochloric Acid → HCID 178472559, compound

HRID 1660431Notes

Reaction equation

HRID 1660431 reaction equation
Cl.[CH3:2][O:3][C:4](=[O:24])[C@H:5]([CH2:17][CH2:18][CH2:19][NH:20][C:21](=[NH:23])[NH2:22])[NH:6][S:7]([C:10]1[CH:16]=[CH:15][C:13]([CH3:14])=[CH:12][CH:11]=1)(=[O:9])=[O:8].P([O-])([O-])([O-])=O.NO.NO.Cl.[OH-].[Na+].[Cl:37][C:38](Cl)(Cl)C(O)=O.Cl>>[ClH:37].[CH2:2]([O:3][C:4](=[O:24])[C@H:5]([CH2:17][CH2:18][CH2:19][NH:20][C:21](=[NH:22])[NH2:23])[NH:6][S:7]([C:10]1[CH:16]=[CH:15][C:13]([CH3:14])=[CH:12][CH:11]=1)(=[O:8])=[O:9])[CH3:38]

Participants

8

Reactant 7

HCID 313Hydrochloric AcidHCID 787HydroxylamineHCID 1061PhosphateHCID 6421Trichloroacetic AcidHCID 14798Sodium HydroxideHCID 443297Hydroxyamine hydrochlorideHCID 178501463HCID 178501463, compound

Product 1

HCID 178472559HCID 178472559, compound

Procedure

To 0.1 ml of thrombin were added 0.3 ml of Nα-tosyl-L-arginine methyl ester hydrochloride solution (10 micromoles/0.4 ml of 5% DMSO) and 0.6 ml of phosphate buffer solution (pH 7.4), and the resulting mixture was subjected to incubation at 37° C. for 30 min, after which 1.5 ml of a hydroxylamine solution (a mixture of equal amounts of 2 M NH2OH hydrochloride and 3.5 M NaOH) was added thereto. The resulting mixture was allowed to stand at room temperature for 15 min. Thereto were added 1 ml of 18% trichloroacetic acid solution, 1 ml of 4 N hydrochloric acid, and 1 ml of 10% ferric chloride solution. The resulting mixture was thoroughly stirred and then centrifuged at 3,000 r.p.m. for 10 min. The color thus developed of the supernatant was measured as absorbance (530 l nm) by means of a spectrophotometer. The value obtained corresponds to the amount of the substrate remaining unhydrolyzed with thrombin, and therefore, and activity of the enzyme corresponds to the difference between the value obtained when no enzyme was used (control) and the value obtained after the enzyme reaction.

Workup

  1. additionwas added
  2. waitto stand at room temperature for 15 min
  3. waitfor 10 min
  4. customThe value obtained corresponds to the amount of the substrate
  5. customactivity of the enzyme corresponds to the difference between the value obtained when no enzyme
  6. customthe value obtained
  7. customafter the enzyme reaction

Sources & evidence

ORD record
ord-d7ca062196934f509a0f21ca41858d1f
Source dataset
uspto-grants-1981_12
Patent number
US04308201

Reaction notes

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