Cholinesterase inhibition based determination of pancuronium bromide in biological samples
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 K-M=0.40 mmol/L, maximal reaction rate V-max=52.2 mu mol/L min, inhibition constant K-i=0,56 mu mol/L and IC50=1.31 mu mol/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 mu mol/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.
Keywords:
pancuronium bromide / cholinesterase inhibition / benzoylcholine / spectrophotometric kinetic determinationSource:
Analytical and Bioanalytical Chemistry, 2006, 385, 8, 1462-1469Publisher:
- Springer Heidelberg, Heidelberg
DOI: 10.1007/s00216-006-0556-5
ISSN: 1618-2642
PubMed: 16804673
WoS: 000239644200017
Scopus: 2-s2.0-33746865535
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Institution/Community
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Stankov-Jovanović, Vesna P. AU - Nikolić-Mandić, Snežana D. AU - Mandić, Ljuba M. AU - Mitić, Violeta D. PY - 2006 UR - https://cherry.chem.bg.ac.rs/handle/123456789/794 AB - 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 K-M=0.40 mmol/L, maximal reaction rate V-max=52.2 mu mol/L min, inhibition constant K-i=0,56 mu mol/L and IC50=1.31 mu mol/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 mu mol/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. PB - Springer Heidelberg, Heidelberg T2 - Analytical and Bioanalytical Chemistry T1 - Cholinesterase inhibition based determination of pancuronium bromide in biological samples VL - 385 IS - 8 SP - 1462 EP - 1469 DO - 10.1007/s00216-006-0556-5 ER -
@article{ author = "Stankov-Jovanović, Vesna P. and Nikolić-Mandić, Snežana D. and Mandić, Ljuba M. and Mitić, Violeta D.", year = "2006", 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 K-M=0.40 mmol/L, maximal reaction rate V-max=52.2 mu mol/L min, inhibition constant K-i=0,56 mu mol/L and IC50=1.31 mu mol/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 mu mol/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.", publisher = "Springer Heidelberg, Heidelberg", journal = "Analytical and Bioanalytical Chemistry", title = "Cholinesterase inhibition based determination of pancuronium bromide in biological samples", volume = "385", number = "8", pages = "1462-1469", doi = "10.1007/s00216-006-0556-5" }
Stankov-Jovanović, V. P., Nikolić-Mandić, S. D., Mandić, L. M.,& Mitić, V. D.. (2006). Cholinesterase inhibition based determination of pancuronium bromide in biological samples. in Analytical and Bioanalytical Chemistry Springer Heidelberg, Heidelberg., 385(8), 1462-1469. https://doi.org/10.1007/s00216-006-0556-5
Stankov-Jovanović VP, Nikolić-Mandić SD, Mandić LM, Mitić VD. Cholinesterase inhibition based determination of pancuronium bromide in biological samples. in Analytical and Bioanalytical Chemistry. 2006;385(8):1462-1469. doi:10.1007/s00216-006-0556-5 .
Stankov-Jovanović, Vesna P., Nikolić-Mandić, Snežana D., Mandić, Ljuba M., Mitić, Violeta D., "Cholinesterase inhibition based determination of pancuronium bromide in biological samples" in Analytical and Bioanalytical Chemistry, 385, no. 8 (2006):1462-1469, https://doi.org/10.1007/s00216-006-0556-5 . .