[The use of enzymatic hydrolysis for isolation of barbituric acid derivatives from blood (as exemplified by phenobarbital and barbamyl)].
Modern isolation techniques by direct extraction with organic solvents or after protein precipitation by various sedimenting or salting-out agents are characterized by low efficiency and do not permit to liberate derivatives of barbituric acid from their complexes with blood proteins. The use of enzymatic hydrolysis makes it possible to break bonds between barbiturates and protein and thereby improve the efficiency of isolation. We performed enzymatic hydrolysis of the model phenobarbital-blood and barbamyl-blood complexes with the use of trypsin, pepsin, chymotrypsin, and papain. The degree of phenobarbital extraction with trypsin and barbamyl was estimated at 62.1 +/- 1.2% and 75.1 +/- 1.6% respectively; in other words, it was 32.7 +/- 1.0% and 51.1 +/- 1.0% higher than that achieved by traditional methods. Certain validation characteristics of the new method are presented.
N A
NA
Chuvina
A S
AS
Kolupaeva
O Iu
OIu
Strelova
I V
IV
Zabolotskaia
T V
TV
Gorbacheva
D000654:Amobarbital / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D001463:Barbiturates / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D001798:Blood Proteins / Q000737:chemistry*; D002918:Chymotrypsin / Q000737:chemistry; D010450:Endopeptidases / Q000737:chemistry*; D005554:Forensic Medicine / Q000379:methods*; D006801:Humans; D006868:Hydrolysis; D006993:Hypnotics and Sedatives / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D010206:Papain / Q000737:chemistry; D010434:Pepsin A / Q000737:chemistry; D010634:Phenobarbital / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D014357:Trypsin / Q000737:chemistry
D001463:Barbiturates; D001798:Blood Proteins; D006993:Hypnotics and Sedatives; D010450:Endopeptidases; D002918:Chymotrypsin; D014357:Trypsin; D010206:Papain; D010434:Pepsin A; D000654:Amobarbital; D010634:Phenobarbital