Levodopa → Dopamine

HRID 543242Notizen

Reaktionsgleichung

Reaktionsgleichung von HRID 543242
[O:1]=[C:2]([C@H:4]([CH2:6][C:7]1[CH:14]=[C:12]([OH:13])[C:10]([OH:11])=[CH:9][CH:8]=1)[NH2:5])[OH:3].C1(C=CC=C(N)C=1O)O>P([O-])([O-])([O-])=O>[NH2:5][CH2:4][CH2:6][C:7]1[CH:8]=[CH:9][C:10]([OH:11])=[C:12]([OH:13])[CH:14]=1.[O:1]=[C:2]([C@H:4]([CH2:6][C:7]1[CH:14]=[C:12]([OH:13])[C:10]([OH:11])=[CH:9][CH:8]=1)[NH2:5])[OH:3]

Komponenten

5

Edukt 2

HCID 6047Levodopa3×HCID 189460Catecholamine

Produkt 2

HCID 681DopamineHCID 6047Levodopa

Reagenzien, Katalysatoren, Lösungsmittel 1

HCID 1061PhosphateLösungsmittel

Versuchsvorschrift

DOPA decarboxylase activity was routinely measured by liquid chromatography and electrochemical detection of catecholamine products. DOPA decarboxylase (1-5 micrograms, 1-20 units) was incubated with L-DOPA (2000 micromolar, 150 microliters) and PLP (10 micromolar) in 0.1M phosphate at pH 6.8 and 37° C. After 1-10 minutes, the reaction was quenched with citric acid (2M, 25 microliters), diluted two-fold with distilled water and 10 microliter aliquots were analyzed by HPLC on a reverse phase Brownlee RP-18 column. Isocratic elution with pH 4.35 buffer [0.1M NaOAc (63% v/v), 0.1M citric acid (32% v/v) and methanol (15% v/v)] at a flow rate of 2.5 ml/min. afforded baseline resolution of dopamine (RT=2.8 min.) from L-DOPA (RT=1.9 min.).

Aufarbeitung

  1. customthe reaction was quenched with citric acid (2M, 25 microliters)
  2. additiondiluted two-fold with distilled water and 10 microliter aliquots
  3. washIsocratic elution with pH 4.35 buffer [0.1M NaOAc (63% v/v), 0.1M citric acid (32% v/v) and methanol (15% v/v)] at a flow rate of 2.5 ml/min.

Quellen und Nachweise

ORD-Eintrag
ord-9c5979020e7040b5810803a55cdc4436
Quelldatensatz
uspto-grants-1987_04
Patentnummer
US04661510

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