TITLE

Cholinesterase inhibition based determination of pancuronium bromide in biological samples

AUTHOR(S)
Stankov-Jovanović, Vesna P.; Nikolić-Mandić, Snežana D.; Mandić, Ljuba M.; Mitić, Violeta D.
PUB. DATE
September 2006
SOURCE
Analytical & Bioanalytical Chemistry;Sep2006, Vol. 385 Issue 8, p1462
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Pancuronium bromide (PCBr) inhibition effect on enzyme cholinesterase from pooled human serum (Che, EC 3.1.1.8 acylcholine acylhydrolase) was used for development of a spectrophotometric kinetic method for PCBr determination in human serum and urine. Optimal conditions for the basic and inhibitor reactions were established: pH=7.7 and substrate concentration c(benzoylcholine chloride)=1.33 mmol/L. Kinetic parameters were also determined: Michaelis-Menten’s constant KM=0.40 mmol/L, maximal reaction rate Vmax=52.2 μmol/L min, inhibition constant Ki=0,56 μmol/L and IC50=1.31 μmol/L. Linear dependence between the reaction rate and inhibitor concentration exists in PCBr concentration range 8.20–68.25 nmol/L, which corresponds to the real sample concentrations from 0.328 to 2.730 μmol/L. The method detection and quantification limits were 2.01 nmol/L and 6.67 nmol/L, respectively. Precision of the method was tested for three pancuronium concentrations (10.70, 29.35 and 51.25 nmol/L). Relative standard deviation (RSD) was in the range 0.15–7.45%. Accuracy was examined by standard addition method. Influence of the substances usually present in serum and urine on the reaction rate was tested. The developed method was applied for PCBr content determination in serum model samples, urine model samples and in urine taken during surgery. The method has good sensitivity, accuracy, precision and it is suitable for clinical practice. [Figure not available: see fulltext.]
ACCESSION #
21857595

 

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