Kinetic study of paracetamol on prolidase activity in erythrocytes by capillary electrophoresis with Ru(bpy)₃²⁺ electrochemiluminescence detection

Abstract

We explored the CE with (bpy) (2+) (3) electrochemiluminescence detection for the kinetic study of drug-enzyme interaction. Effects of four nonsteroidal anti-inflammatory drugs including aspirin, paracetamol, sodium salicylate and phenacetin on prolidase (PLD) activity in erythrocytes were investigated. Aspirin enhanced PLD activity whereas the other three had inhibiting effects. This may reveal their different effects on the collagen biosynthesis and catabolism that influence tumor invasiveness. Kinetic study of paracetamol on PLD showed that the value of Michaelis constant K(m) for PLD was 1.23 mM. The mechanism of PLD inhibition by paracetamol is noncompetitive inhibition, and the inhibitor constant K(i) value obtained in our research was 9.73 x 10(3) microg/L.

Electrogenerated Chemiluminescence and Its Biorelated Applications

“…The CE-ECL technology also has been applied for the thermodynamic and kinetic studies, for example, of drug–human serum albumin (BSA) binding, , of paracetamol on prolidase activity in erythrocytes, and in the serum of diabetic patients, with procain hydrolysis as a probe for butyrylcholinesterase by in vitro procain metabolism in plasma with butyrylcholinesterase acting as bioscavenger, and the interaction of chloroquine phosphate with BSA . A number of advantages are offered by CE-ECL with respect to other commonly used separation−detection methods such as HPLC using UV−vis or fluorescent detection for this kind of study and include very powerful resolving ability, good selectivity, high sensitivity, easy sample preparation, and fast data measurement as generally no sample pretreatment is needed and no interference exists. ,, Characterization of high-affinity antibodies and rapid estimation of enzyme activity in the hydrolysis of peptides by flow-cell-based Ru(bpy) 3 2+ ECL have also been recently demonstrated.…”

Ruthenium Polypyridine Complexes Combined with Oligonucleotides for Bioanalysis: A Review

“…Derived from the simple and inexpensive platform offered by electrochemical DNA sensing, a more sensitive analytical strategy has been proposed, namely, the ECL-based DNA detection. ECL based on Ru(bpy) 3 2+ and its analogues has been widely used in detection of many important analytes including metal ions, bioactive small molecules, drugs, protein, DNA, bacteria, cancer cells and so on with high selectivity and sensitivity [65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97,98,99,100,101,102,103,104,105,106,107,108,109,110,111,112,113,114,115,116,117,118,119,120,121,122,123,124,125,…”

Advances and analytical applications in chemiluminescence coupled to capillary electrophoresis

“…(Table 1). A relevant application consisted on the determination of proline produced from enzymatic reaction and its application to assay prolidase activity in nonpretreated serum of diabetic patients to evaluate collagen degradation in diabetes mellitus [55] or to study drug-enzyme interactions, such as non-steroidal anti-inflammatory drugs on prolidase activity in erythrocytes [56]. Other innovative contributions carried out with this commercial device include the application of ECL detection for the chiral separation of a racemic mixture of disopyramide in plasma, in which the buffers used in the capillary inlet and outlet differed from each other in terms of buffer pH, ionic strength, type of BGE as well as buffer composition [57], or the use of ionic liquid as an additive of the running buffer for the separation of main opium alkaloids in Chinese traditional medicine, providing high sensitivity and good reproducibility [58].…”